Note: Descriptions are shown in the official language in which they were submitted.
PSU004-CADIV1
DEPHOSPHORYLATION-CONTROLLED EXTRACTION OF PHOSPHORYLATABLE
PSYCHOACTIVE ALKALOIDS
TECHNICAL FIELD
[1] This application relates to a process of obtaining psychoactive
alkaloid
extracts. More specifically, the present invention relates to controlling
dephosphorylation during extraction. Further, the present invention relates to
psychoactive alkaloid compositions with controlled dephosphorylation.
BACKGROUND
[2] Varieties of mushrooms have played important roles in most societies.
The
active ingredients in mushrooms have been found to have medicinal properties
including relief of symptoms of various diseases and conditions. The
concentration of
active ingredients for these applications may vary not only from species to
species, but
also from mushroom to mushroom inside a given species, subspecies, or variety.
The
same holds true even for different parts of the same mushroom or mycelium.
[3] Various methods of extraction, which have been used to separate natural
extracts from a variety of mushrooms, have resulted in difficulties with large
crop-to-
crop variability. A large variability within a single plant or fungus
sometimes causes
inconsistent concentration of the active psychoactive compound and its
stability.
Different solvent choices extract the psychoactive compounds equally, some of
them
electively extract one or the other, and some convert the compounds between
each
other or degrade them into non-psychoactive compounds. Many extraction
processes
for extracting standardized concentrations of the compounds for direct medical
use are
usually complex. This results in expensive extraction processes and a high
cost of
isolated, natural extracts.
[4] U.S. Patent 3183172 to Heim et al. relates to an industrial process for
the
isolation of active compounds from mushrooms grown under predetermined
conditions.
With the predetermined growing conditions, mushrooms grew with ten times more
active mycelium and sclerotium and increased concentrations of psychoactive
compounds. However, a large portion of the target compounds were lost during
the
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PSU004-CADIV1
extraction process or not extracted at all. In addition, the solvent and
solvent systems
used during the extraction process were materials such as methanol, acetone,
dichloromethane, diethyl ether, or others known to be toxic to humans, even in
small
quantities.
[5] Extraction methods currently described in the art are inefficient such
that a
large portion of the target compounds were lost during the extraction process
or not
extracted at all. They lack analytical data as evidence. We have replicated
and
measured these methods to prove they are ineffective. In addition, they
describe the
use of solvents which are known to be toxic to humans, even in small
quantities.
[6] To date, the focus has largely been on synthetic preparations of these
compounds because of the many difficulties associated with naturally extracted
preparations. It is currently infeasible and expensive to extract psilocybin
from
mushrooms, and even the best chemical synthesis methods require expensive and
difficult-to-source starting substrates.
[7] Accordingly, there is a need of methods to produce high efficiency,
standardized preparations of the target compounds for medical use while using
acceptable solvent systems to create a more consistent supply chain.
SUMMARY OF INVENTION
[8] A psychoactive alkaloid composition comprising of, by weight: 0.1-99.9%
of a
psychoactive alkaloid extract; and one or more preservatives up to 10%, a flow
agent
up to 2%, 0-94% of one or more carriers, or any combination thereof. In some
embodiments, the composition comprises 2-99.7% of the psychoactive alkaloid
extract.
In some embodiments, the composition comprises an antioxidant up to 0.5% by
weight.
In some embodiments, the composition comprises a bioavailability agent up to
0.5% by
weight. In some embodiments, the psychoactive alkaloid extract has a
psychoactive
alkaloid concentration ranging from 0.1`)/0 to 99% by weight of the extract.
In some
embodiments, the one or more preservatives are selected from ascorbic acid,
citric
acid, lactose, vitamin A, vitamin E, retinyl palmitate, selenium, sodium
citrate, sodium
ascorbate, calcium ascorbate, sodium benzoate, and potassium benzoate. In some
embodiments, the psychoactive alkaloid extract has a psychoactive alkaloid
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concentration ranging from 0.1% to 20% by weight of the extract. In some
embodiments, the psychoactive alkaloid extract is a purified psychoactive
alkaloid
extract, and the purified psychoactive alkaloid extract has a psychoactive
alkaloid
concentration ranging from 10% to 99% by weight. In some embodiments, the
composition is in a powder form. In some embodiments, the composition
comprises
10% or more of the carrier. In some embodiments, the psychoactive alkaloid is
psilocybin, psilocin, baeocystin, norbaeocystin, norpsilocin, aeruginascin,
bufotenin,
bufotenidine, 5-Me0-DMT (5-methoxy-N.Ndimethyltryptamine), N,N-
dimethyltryptamine
(DMT), 4-hydroxytryptamine, N,N,N-trimethy1-4-hydroxytryptamine ergine (LSA),
ergonovine, ergometrine, muscimol, ibotenic acid, lysergic acid
hydroxyethylamide
(LSH), elymoclavine, ergometrinine, and/or chanoclavine, or any combination
selected
therefrom. In some embodiments, the psychoactive alkaloid extract comprises
naturally
occurring substances selected from the group consisting of fats, sugars,
carbohydrates,
chitin, chitosan, minerals, waxes and proteins. In some embodiments, the
naturally
occurring substances are present in the psychoactive alkaloid extract in a
concentration
ranging from 1% - 99.9% by weight. In some embodiments, the psychoactive
alkaloid
extract is from fungi. In some embodiments, the psychoactive alkaloid extract
is from
Psilocybe cyanescens, Psilocybe cubensis, Amanita muscaria, or any selection
therefrom. In some embodiments, the psychoactive alkaloid extract is from
psychoactive plants. In some embodiments, the psychoactive alkaloid extract is
from
Anadenanthera colubrina. In some embodiments, the psychoactive alkaloid
extract is
from Anadenanthera peregrina. In some embodiments, the psychoactive alkaloid
extract is from psychoactive animals. In some embodiments, the psychoactive
alkaloid
extract is from Incilius alvarius. In some embodiments, the psychoactive
alkaloid
extract is from psychoactive yeasts. In some embodiments, the flow agent is
selected
from silicon dioxide, stearic acid, magnesium stearate, or talc. In some
embodiments,
the one or more carriers are selected from starch, maltodextrin, alpha and
beta
cyclodextrin, microcrystalline cellulose (MCC), gum arabic, xanthum gum, guar
gum,
mannitol, or cellulose gum. In some embodiments, the maltodextrin is tapioca
maltodextrin or rice maltodextrin. In some embodiments, the starch is potato
starch. In
some embodiments, the flow agent is present in the composition at 0.1 to 1.2%.
In
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PSU004-CADIV1
some embodiments, a first preservative of the one or more preservatives is
present in
the composition at 0.1 to 3%. In some embodiments, a second preservative of
the one
or more preservatives is present in the composition at 0.1 to 3%. In some
embodiments, a first carrier of the one or more carriers is present in the
composition at
to 20%. In some embodiments, a second carrier of the one or more carriers is
present in the composition at 10 to 20%. In some embodiments, the composition
comprises: the flow agent present in the composition at 0.1 to 1.2%; the one
or more
preservatives present in the composition at 0.1 to 2%; and the one or more
carriers
present in the composition at 10 to 20%. In some embodiments, the flow agent
is
silicon dioxide; the carrier comprises maltodextrin and mannitol; and the one
or more
preservatives comprise ascorbic acid and citric acid. In some embodiments, the
silicon
dioxide is present in the composition at 0.1 to 1.2%. In some embodiments, the
ascorbic acid is present in the composition at 0.1 to 2%. In some embodiments,
the
citric acid is present in the composition at 0.1 to 2%. In some embodiments,
the
maltodextrin is present in the composition at 10 to 20%. In some embodiments,
the
mannitol is present in the composition at 10 to 20%. In some embodiments, the
flow
agent is silicon dioxide; the carrier comprises starch and mannitol; and the
one or more
preservatives comprise ascorbic acid and citric acid. In some embodiments, the
silicon
dioxide is present in the composition at 0.1 to 1.2%. In some embodiments, the
ascorbic acid is present in the composition at 0.1 to 2%. In some embodiments,
the
citric acid is present in the composition at 0.1 to 2%. In some embodiments,
the starch
is present in the composition at 10 to 20%. In some embodiments, the mannitol
is
present in the composition at 10 to 20%.
[9] A
method for generating a psychoactive alkaloid extract from a psychoactive
organism, the method comprising: providing a biomass of the psychoactive
organism;
contacting the biomass with 10 to 100 milliliters (mL) of solvent per gram (g)
of the
biomass; and evaporating the solvent from the biomass to yield the
psychoactive
alkaloid extract. In some embodiments, the solvent is selected from 100%
methanol, an
alcohol:water mixture wherein the alcohol comprises 60% to 99% of the
alcohol:water
mixture, an alcohol:acid mixture wherein the alcohol comprises 60% to 99% of
the
alcohol:acid mixture, and acidified water.
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[10] A
method for generating a psychoactive alkaloid extract from a psychoactive
organism, the method comprising: providing a biomass of the psychoactive
organism;
contacting the biomass with a solvent, wherein the solvent is selected from
100%
methanol, an alcohol:water mixture wherein the alcohol comprises 60% to 99% of
the
alcohol:water mixture, an alcohol:acid mixture wherein the alcohol comprises
60% to
99% of the alcohol:acid mixture, and acidified water; and evaporating the
solvent from
the biomass to yield the psychoactive alkaloid extract. In some embodiments,
the
solvent is present at 10 to 100 milliliters (mL) of solvent per gram (g) of
the biomass. In
some embodiments, the solvent is present at 10 to 60 mL of solvent per gram of
the
biomass. In some embodiments, the solvent is present at 40 to 60 mL of solvent
per
gram of the biomass. In some embodiments, the biomass of the psychoactive
organism
is dried prior to contacting with the solvent. In some embodiments, the
biomass of the
psychoactive organism is reduced to a particle size of 6 millimeters (mm) to
0.03 mm
prior to contacting with the solvent. In some embodiments, the biomass of the
psychoactive organism is reduced to a particle size of 1 mm to 0.03 mm prior
to
contacting with the solvent. In some embodiments, the biomass of the
psychoactive
organism is reduced to a particle size of at least 0.074 mm prior to
contacting with the
solvent. In some embodiments, the biomass of the psychoactive organism is
contacted
with the solvent at 5 C to 95 C. In some embodiments, the biomass of the
psychoactive
organism is contacted with the solvent at 20 C to 70 C. In some embodiments,
the
biomass of the psychoactive organism is contacted with the solvent at 25 C. In
some
embodiments, the biomass of the psychoactive organism is contacted with the
solvent
for 1 to 720 minutes. In some embodiments, the biomass of the psychoactive
organism
is contacted with the solvent for 20 to 60 minutes. In some embodiments, the
biomass
of the psychoactive organism is contacted with the solvent for 30 minutes. In
some
embodiments, following (b), the biomass is filtered through a filter. In some
embodiments, the filter comprises a 1 micron (pm) to 10 pm mesh. In some
embodiments, following (b), the biomass is contracted with a second solvent.
In some
embodiments, the second solvent is selected from 100% methanol, an
alcohol:water
mixture wherein the alcohol comprises 60% to 99% of the alcohol:water mixture,
an
alcohol:acid mixture wherein the alcohol comprises 60% to 99% of the
alcohol:acid
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PSU004-CADIV1
mixture, and acidified water. In some embodiments, the second solvent is
present at 10
to 100 milliliters (mL) of solvent per gram (g) of the biomass. In some
embodiments, the
solvent is present at 10 to 60 mL of solvent per gram of the biomass. In some
embodiments, the solvent is present at 40 to 60 mL of solvent per gram of the
biomass.
In some embodiments, the alcohol of the alcohol:water mixture, the
alcohol:acid
mixture, or both, is a C1-C4 primary aliphatic alcohol. In some embodiments,
the Cl-C4
primary aliphatic alcohol is ethanol or methanol. In some embodiments, the
acid in the
alcohol:acid mixture, the acidified water, or both, is acetic acid, adipic
acid, ascorbic
acid, phosphoric acid, ammonium aluminum sulphate, ammonium citrate dibasic,
ammonium citrate monobasic, calcium citrate, calcium fumarate, calcium
gluconate,
calcium phosphate dibasic, calcium phosphate, hydrochloric acid, sulphuric
acid
monobasic, calcium phosphate tribasic, citric acid, fumaric acid, gluconic
acid,
magnesium fumarate, malic acid, phosphoric acid, potassium acid tartrate,
potassium
citrate, potassium fumarate, sodium citrate, sodium fumarate, sodium
gluconate,
sodium lactate, sodium potassium hexametaphosphate, sodium potassium tartrate,
sodium potassium tripolyphosphate, sodium pyrophosphate tetrabasic, sodium
tripolyphosphate, tartaric acid, and any combination therefrom. In some
embodiments,
the solvent, second solvent, or both is buffered to a pH of either 4 or less,
or 10 or
greater. In some embodiments, the solvent, second solvent, or both is buffered
with
ammonium bicarbonate, ammonium carbonate, ammonium hydroxide, calcium acetate,
calcium carbonate, calcium chloride, calcium hydroxide, calcium lactate,
calcium oxide,
calcium phosphate, dibasic, calcium phosphate monobasic, magnesium carbonate,
potassium aluminum sulphate, potassium bicarbonate, potassium carbonate,
potassium
hydroxide, potassium lactate, potassium phosphate, dibasic, potassium
pyrophosphate,
tetrabasic, potassium phosphate tribasic, potassium tripolyphosphate, sodium
acetate,
sodium acid pyrophosphate, sodium aluminum phosphate, sodium aluminum
sulphate,
sodium bicarbonate, sodium bisulphate, sodium carbonate, sodium
hexametaphosphate, sodium hydroxide, sodium lactate, sodium phosphate dibasic,
sodium phosphate monobasic, sodium phosphate tribasic, and any combination
therefrom. In some embodiments, the solvent, second solvent, or both is
buffered with
acetic acid, adipic acid, ascorbic acid, phosphoric acid, ammonium aluminum
sulphate,
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ammonium citrate dibasic, ammonium citrate monobasic, calcium citrate, calcium
fumarate, calcium gluconate, calcium phosphate dibasic, calcium phosphate,
hydrochloric acid, sulphuric acid monobasic, calcium phosphate tribasic,
citric acid,
fumaric acid, gluconic acid, magnesium fumarate, malic acid, phosphoric acid,
potassium acid tartrate, potassium citrate, potassium fumarate, sodium
citrate, sodium
fumarate, sodium gluconate, sodium lactate, sodium potassium
hexametaphosphate,
sodium potassium tartrate, sodium potassium tripolyphosphate, sodium
pyrophosphate
tetrabasic, sodium tripolyphosphate, tartaric acid, and any combination
therefrom. In
some embodiments, the psychoactive alkaloid is psilocybin, psilocin,
baeocystin,
norbaeocystin, norpsilocin, aeruginascin, bufotenin, bufotenidine, 5-Me0-DMT
(5-
methoxy-N.Ndimethyltryptamine), N,N-dimethyltryptamine (DMT), 4-
hydroxytryptamine,
N,N,N-trimethy1-4-hydroxytryptamine ergine (LSA), ergonovine, ergometrine,
muscimol,
ibotenic acid, lysergic acid hydroxyethylamide (LSH), elymoclavine,
ergometrinine,
and/or chanoclavine, or any combination selected therefrom. In some
embodiments, the
solvent has a pH of 10 or greater and the psychoactive alkaloid extract
comprises
greater than 50% of the phosphorylated psychoactive alkaloid. In some
embodiments,
the psychoactive alkaloid extract comprises greater than 90% of a
phosphorylated
psychoactive alkaloid. In some embodiments, the phosphorylated alkaloid is
psilocybin,
baeocystin, norbaeocystin, aeruginascin, or any combination therefrom. In some
embodiments, the solvent has a pH of 4 or less and the psychoactive alkaloid
extract
comprises greater than 50% of a dephosphorylated psychoactive alkaloid. In
some
embodiments, the psychoactive alkaloid extract comprises greater than 90% of
the
dephosphorylated psychoactive alkaloid. In some embodiments, the
dephosphorylated
alkaloid is psilocin, norpsilocin, 4-hydroxytryptamine, N,N,N-trimethy1-4-
hydroxytryptamine, or any combination therefrom. In some embodiments, the
psychoactive organism is a plant, animal, fungus, protist, or bacterium. In
some
embodiments, the psychoactive organism is Psilocybe cyanescens, Psilocybe
cubensis,
Amanita muscaria, or any selection therefrom. In some embodiments, the
psychoactive
organism is Anadenanthera colubrina or Anadenanthera peregrina. In some
embodiments, the psychoactive organism is Incilius alvarius. In some
embodiments, the
psychoactive organism is yeast.
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[11] A process for forming an extract of psychoactive alkaloids from
psychoactive
organisms comprising the steps of: soaking a biomass of dried, raw psychedelic
fungus
in a solvent selected from the group consisting of ethanol, a water-ethanol
mixture,
methanol, and a water-methanol mixture in order to dissolve the psychoactive
alkaloids
in the solvent; filtering an undissolved portion of the biomass from the
solvent;
evaporating the solvent sufficiently to remove the solvent completely, leaving
a
concentrated slurry or a residue that is converted to the concentrated slurry
by adding
water thereto: and standardizing the concentrated slurry by adding thereto a
quantity of
carrier measured to achieve a specified purity of extract. In some
embodiments, the
standardizing comprises: measuring a psychoactive alkaloid content in the
concentrated slurry; and using the psychoactive alkaloid content, the
specified purity
and a volume of the concentrated slurry to determine the quantity of carrier.
In some
embodiments, the process comprises drying the concentrated slurry to result in
the
extract, wherein the extract is a powdered extract. In some embodiments, the
solvent is
a water-ethanol or water-methanol alkaline buffered solution. In some
embodiments,
the solvent has a pH of 11-12. In some embodiments, the solvent is buffered
with
sodium hydroxide, the process comprising, between the filtering and
evaporating steps,
adjusting the solvent to a pH of 4-9 using phosphoric acid. In some
embodiments, the
solvent is a water-ethanol or water-methanol acid buffered solution. In some
embodiments, the solvent has a pH of 1.8-3. In some embodiments, the solvent
is
buffered with citric acid, the process comprising, between the filtering and
evaporating
steps, adjusting the solvent to a pH of 4-9 using sodium hydroxide. In some
embodiments, the solvent comprises 100% reverse osmosis water. In some
embodiments, the soaking is at a temperature of 5-95 C. In some embodiments,
the
process comprises applying a pressure of 50 kPa ¨ 100 MPa to the solvent
during the
soaking step. In some embodiments, the process comprises agitating the solvent
during
the soaking step, wherein the soaking step has a duration of 10 minutes to 12
hours. In
some embodiments, the psychedelic organism is a plant, animal, fungus,
protist, or
bacterium. In some embodiments, the fungus comprises Amanita muscaria,
Psilocybe
cubensis, Psilocybe cyanescens, or any combination thereof. In some
embodiments,
the psychoactive alkaloids comprise psilocybin, psilocin, baeocystin,
norbaeocystin,
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ibotenic acid or any mixture thereof. In some embodiments, the solvent to
biomass ratio
is in a range from 1L:lkg to 50L:1kg. In some embodiments, the specified
purity is 0.1-
10%. In some embodiments, the specified purity is specified as a percentage
with a
precision of two decimal places. In some embodiments, the carrier comprises
ascorbic
acid, silicon dioxide, maltodextrin, gum arabic, microcrystalline cellulose,
sodium citrate,
sodium benzoate, sodium phosphate, rice, rice hulls, or any combination of the
foregoing. In some embodiments, the process comprises=repeating, using further
solvent, the soaking and filtering steps for the filtered biomass; and
combining the
filtered solvent with the filtered further solvent.
[12] A process for obtaining a purified psychoactive alkaloid solution, the
process
comprising: extracting a psychoactive alkaloid from a psychoactive alkaloid
source to
obtain a psychoactive alkaloid extract; contacting the psychoactive alkaloid
extract with
an adsorbent material to obtain an adsorbed psychoactive alkaloid; and eluting
the
adsorbed psychoactive alkaloid using a solvent to obtain a purified
psychoactive
alkaloid solution, wherein the solvent is water, an organic solvent or a
combination
thereof, under basic, acidic or neutral pH. In some embodiments, the process
comprises prior to the treating step, adding an acid or a base to the
psychoactive
alkaloid extract. In some embodiments, after adding the acid or base, the
psychoactive
alkaloid extract has a pH ranging from 2.5-4.5 or from 9-10 respectively. In
some
embodiments, the acid is selected from the group consisting of acetic acid,
adipic acid,
ascorbic acid, phosphoric acid, ammonium aluminum sulphate, ammonium citrate
dibasic, ammonium citrate monobasic, calcium citrate, calcium fumarate,
calcium
gluconate, calcium phosphate dibasic, calcium phosphate, hydrochloric acid,
sulphuric
acid monobasic, calcium phosphate tribasic, citric acid, fumaric acid,
gluconic acid,
magnesium fumarate, malic acid, phosphoric acid, potassium acid tartrate,
potassium
citrate, potassium fumarate, sodium citrate, sodium fumarate, sodium
gluconate,
sodium lactate, sodium potassium hexametaphosphate, sodium potassium tartrate,
sodium potassium tripolyphosphate, sodium pyrophosphate tetrabasic, sodium
tripolyphosphate, tartaric acid, and any combination therefrom. In some
embodiments,
the base is selected from the group consisting of ammonium bicarbonate,
ammonium
carbonate, ammonium hydroxide, calcium acetate, calcium carbonate, calcium
chloride,
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calcium hydroxide, calcium lactate, calcium oxide, calcium phosphate, dibasic,
calcium
phosphate monobasic, magnesium carbonate, potassium aluminum sulphate,
potassium bicarbonate, potassium carbonate, potassium hydroxide, potassium
lactate,
potassium phosphate, dibasic, potassium pyrophosphate, tetrabasic, potassium
phosphate tribasic, potassium tripolyphosphate, sodium acetate, sodium acid
pyrophosphate, sodium aluminum phosphate, sodium aluminum sulphate, sodium
bicarbonate, sodium bisulphate, sodium carbonate, sodium hexametaphosphate,
sodium hydroxide, sodium lactate, sodium phosphate dibasic, sodium phosphate
monobasic, sodium phosphate tribasic, and any combination therefrom. In some
embodiments, the adsorbent material is a gel resin, a macroporous resin, or a
combination thereof. In some embodiments, the macroporous resin is a non-ionic
macroporous resin, an ion-exchange macroporous resin, or a combination
thereof. In
some embodiments, the psychoactive alkaloid source comprises psilocybin,
psilocin,
baeocystin, norbaeocystin, norpsilocin, aeruginascin, bufotenin, bufotenidine,
5-Me0-
DMT (5-methoxy-N.N-dimethyltryptamine), N,N-dimethyltryptamine (DMT), ergine
(LSA), ergonovine, ergometrine, muscimol, ibotenic acid, lysergic acid
hydroxyethylamide (LSH), elymoclavine, ergometrinine, chanoclavine, or any
combination therefrom. In some embodiments, the organic solvent is selected
from a
group consisting of C1-4 primary aliphatic alcohols, C3-4 ketones, and any
combination
therefrom. In some embodiments, the process comprises further purifying the
obtained
purified psychoactive alkaloid solution by repeating, with the obtained
purified
psychoactive alkaloid solution, the treating step with a different adsorbent
material and
the eluting step with another solvent. In some embodiments, the process
comprises
evaporating a portion of solvent from the purified psychoactive alkaloid
solution to
obtain a purified psychoactive alkaloid slurry. In some embodiments, the
purified
psychoactive alkaloid slurry comprises 5% by weight or more of a psychoactive
alkaloid. In some embodiments, the process comprises standardizing the
purified
psychoactive alkaloid slurry by adding thereto a quantity of excipient
measured to
provide a specific concentration of psychoactive alkaloid when the purified
psychoactive
alkaloid slurry is dried; and drying the purified psychoactive alkaloid slurry
by
evaporating the remaining portion of the solvent to obtain a standardized,
purified,
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powdered psychoactive alkaloid extract having the specific concentration of
psychoactive alkaloid. In some embodiments, the psychoactive alkaloid
comprises
psilocybin, psilocin, baeocystin, norbaeocystin, norpsilocin, aeruginascin,
bufotenin,
bufotenidine, 5-Me0-DMT (5-methoxy-N.N-dimethyltryptamine), N,N-
dimethyltryptamine (DMT), ergine (LSA), ergonovine, ergometrine, muscimol,
ibotenic
acid, lysergic acid hydroxyethylamide (LSH), elymoclavine, ergometrinine,
chanoclavine, or any combination therefrom; and the standardized, purified,
powdered
psychoactive alkaloid extract has a psychoactive alkaloid concentration
ranging from
0.1-99% by weight. In some embodiments, the excipient is selected from the
group
consisting of silicon dioxide, ascorbic acid, maltodextrin, gum arabic,
microcrystalline
cellulose, sodium benzoate, sodium phosphate, sodium citrate, rice hulls, rice
and any
combination therefrom. In some embodiments, the process comprises prior to the
treating step: adding an acid to the psychoactive alkaloid extract to bring
its pH to
4 0.5; and removing solids from the psychoactive alkaloid extract; and after
the treating
step and before the eluting step: washing the adsorbent material with purified
water;
wherein: the adsorbent material is a non-ionic macroporous resin; and the
solvent used
for the eluting step is a hydro-ethanol solvent. In some embodiments, the
hydro-
ethanol solvent is 5% ethanol. In some embodiments, the process comprises
prior to
the treating step: adding an acid to the psychoactive alkaloid extract to
bring its pH to
3 0.5; after the treating step and before the eluting step: washing the
adsorbent
material with 100% ethanol, wherein the adsorbent material is a macroporous
strong
cation exchange resin in an H+ or an Na + form; and washing the adsorbent
material with
purified water; wherein the solvent used for the eluting step is 2%
hydrochloric acid and
80% ethanol; and after the eluting step: adding alkali to the purified
psychoactive
alkaloid solution to bring its pH to 4 0.5; removing solids from the purified
psychoactive
alkaloid solution; evaporating a portion of the solvent from the purified
psychoactive
alkaloid solution; removing further solids from the purified psychoactive
alkaloid
solution; treating the purified psychoactive alkaloid extract with a non-ionic
macroporous resin to obtain a second adsorbed psychoactive alkaloid; washing
the
non-ionic macroporous resin with purified water; and eluting the second
adsorbed
psychoactive alkaloid from the non-ionic macroporous resin using a hydro-
ethanol
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solvent to obtain a twice purified psychoactive alkaloid solution. In some
embodiments,
the process comprises prior to the treating step: adding a base to the
psychoactive
alkaloid extract to bring its pH to 9.5 0.5; after the treating step and
before the eluting
step: washing the adsorbent material with 100% ethanol, wherein the adsorbent
material is a macroporous strong anion exchange resin in an OH- or a Cl- form;
and
washing the adsorbent material with purified water; wherein the solvent used
for the
eluting step is 2% sodium chloride and 80% ethanol; and after the eluting
step: adding
acid to the purified psychoactive alkaloid solution to bring its pH to 4 0.5;
removing
solids from the purified psychoactive alkaloid solution; evaporating a portion
of the
solvent from the purified psychoactive alkaloid solution; removing further
solids from the
purified psychoactive alkaloid solution; treating the purified psychoactive
alkaloid extract
with a non-ionic macroporous resin to obtain a second adsorbed psychoactive
alkaloid;
washing the non-ionic macroporous resin with purified water; and eluting the
second
adsorbed psychoactive alkaloid from the non-ionic macroporous resin using a
hydro-
ethanol solvent to obtain a twice purified psychoactive alkaloid solution. In
some
embodiments, the psychoactive alkaloid source comprises psychoactive fungus
and the
extracting step comprises: drying and pulverizing the psychoactive alkaloid
source to
obtain a dried biomass; heating the dried biomass in a first solvent to obtain
a first
slurry, and filtering the first slurry to obtain a first filtrate and a first
residue; heating the
first residue in a second solvent to obtain a second slurry, and filtering the
second slurry
to obtain a second filtrate and a second residue; and mixing the first
filtrate and the
second filtrate to obtain the psychoactive alkaloid extract. In some
embodiments, the
first solvent and the second solvent are selected from a group consisting of a
primary
aliphatic alcohol, a ketone, purified water, and any combination therefrom;
and the
heating is carried out at a temperature ranging from 5-95 C and for a time
duration
ranging from 10 minutes to 12 hours. In some embodiments, the psychoactive
alkaloid
source is Anadenanthera peregrina, the process comprising: prior to the
treating step:
adding an acid to the psychoactive alkaloid extract to bring its pH to 4 0.5;
and
removing solids from the psychoactive alkaloid extract; and after the treating
step and
before the eluting step: washing the adsorbent material with purified water
then with
10% ethanol; wherein: the adsorbent material is a macroporous resin; and the
solvent
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used for the eluting step is 50% ethanol. In some embodiments, the
psychoactive
alkaloid source comprises a plant, animal, fungus, protist, or bacterium. In
some
embodiments, the psychoactive alkaloid source cornprises Psilocybe cyanescens,
Psilocybe cubensis, Amanita muscaria, or any selection therefrom. In some
embodiments, the psychoactive alkaloid source comprises Anadenanthera
colubrina or
Anadenanthera peregrina. In some embodiments, the psychoactive alkaloid source
comprises Incilius alvarius. In some embodiments, the psychoactive alkaloid
source
comprises yeast. In some embodiments, the psychoactive alkaloid extract is
contacted
with the adsorbent material at a flow rate of 1 bed volume per hour (BV/h) to
10 BV/h. In
some embodiments, the psychoactive alkaloid extract is contacted with the
adsorbent
material at a flow rate of 2 bed volumes per hour (BV/h) to 6 BV/h.
[13] A
process for obtaining a psychoactive alkaloid extract with a desired amount
of a phosphorylated psychoactive alkaloid and a desired amount of a
dephosphorylated
psychoactive alkaloid, the process comprising: drying and pulverizing a
psychoactive
alkaloid source to obtain a dried powdered biomass; extracting a psychoactive
alkaloid
from the dried powdered biomass with an acidified solvent or a basified
solvent to
obtain a psychoactive alkaloid liquid with a specific pH, wherein the specific
pH is lower
than 3.5 or greater than 10.5; adjusting the pH of the psychoactive alkaloid
liquid to a
pH ranging from 3.5-4.5; and evaporating the solvent from the psychoactive
alkaloid
liquid to obtain the psychoactive alkaloid extract with the desired amount of
the
phosphorylated psychoactive alkaloid and the desired amount of the
dephosphorylated
psychoactive alkaloid; wherein: the desired amount of the phosphorylated
psychoactive
alkaloid is 0-100% by weight of a total phosphorylatable psychoactive alkaloid
content
in the psychoactive alkaloid extract; and the desired amount of the
dephosphorylated
psychoactive alkaloid is the remainder of the total phosphorylatable
psychoactive
alkaloid content in the psychoactive alkaloid extract. In some embodiments,
the
phosphorylated alkaloid is psilocybin, baeocystin, norbaeocystin,
aeruginascin, or any
combination therefrom; and the dephosphorylated alkaloid is psilocin,
norpsilocin, 4-
hydroxytryptamine, N,N,N-trimethy1-4-hydroxytryptamine, or any combination
therefrom.
In some embodiments, the psychoactive alkaloid source comprises psilocybin,
baeocystin, norbaeocystin, aeruginascin, psilocin, norpsilocin, 4-
hydroxytryptamine,
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N,N,N-trimethy1-4-hydroxytryptamine, or any combination therefrom. In some
embodiments, the extracting step comprises: mixing the dried powdered biomass
with
the acidified solvent or the basified solvent to obtain a slurry; and
filtrating the slurry to
obtain a filtrate residue and the psychoactive alkaloid liquid. In some
embodiments, the
extracting step comprises further extracting the psychoactive alkaloid by
repeating, with
the obtained filtrate residue, the extracting step with the same or a
different acidified
solvent, or the same or a different basified solvent. In some embodiments,
after the
mixing step the acidified solvent or the basified solvent, the slurry has a pH
ranging
from 0.5-3.5 or from 10.5-13.5 respectively. In some embodiments, the
acidified solvent
is a mixture of an acid; and a C1-C4 primary aliphatic alcohol, a C3-C4
ketone, water,
or any combination selected therefrom. In some embodiments, the basified
solvent is a
mixture of a base; and a C1-C4 primary aliphatic alcohol, a C3-C4 ketone,
water, or any
combination selected therefrom. In some embodiments, the extraction is
performed: at
a temperature ranging from 5-95 C; and for a time period ranging from 10-720
minutes.
In some embodiments, the extraction is performed at a pressure ranging from 50
kPa ¨
138 MPa (7 to 20,000 psi). In some embodiments, the extraction is performed
with a
solvent to solid proportion of 1L:1kg to 50L:1kg, wherein the solid is the
dried powdered
biomass. In some embodiments, the specific pH is lower than 3.5 and the
desired
amount of the phosphorylated psychoactive alkaloid is 0% by weight of the
total
phosphorylatable psychoactive alkaloid content in the psychoactive alkaloid
extract,
and the desired amount of the dephosphorylated psychoactive alkaloid is 100%
by
weight of the total phosphorylatable psychoactive alkaloid content in the
psychoactive
alkaloid extract. In some embodiments, the specific pH is greater than 10.5
and the
desired amount of the phosphorylated psychoactive alkaloid is 100% by weight
of the
total phosphorylatable psychoactive alkaloid content in the psychoactive
alkaloid
extract, and the desired amount of the dephosphorylated psychoactive alkaloid
is 0%
by weight of the total phosphorylatable psychoactive alkaloid content in the
psychoactive alkaloid extract. In some embodiments, the specific pH is greater
than
10.5, and a maximum desired amount of the phosphorylated alkaloid is limited
by an
amount of the dephosphorylated alkaloid in the psychoactive alkaloid source.
In some
embodiments, the specific pH is greater than 10.5, and the desired amount of
the
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phosphorylated psychoactive alkaloid is 1-99% by weight of the total
phosphorylatable
psychoactive alkaloid content in the psychoactive alkaloid extract. In some
embodiments, the process comprises pausing the evaporating step when a portion
of
the solvent has been evaporated from the psychoactive alkaloid liquid to
obtain a
psychoactive alkaloid slurry; standardizing the psychoactive alkaloid slurry
by adding
thereto a measured quantity of one or more excipients to obtain a standardized
slurry
with a specific amount of psychoactive alkaloid content; and continuing the
evaporating
step by drying the standardized slurry to obtain a psychoactive alkaloid
composition
comprising the psychoactive alkaloid extract, and one or more excipients;
wherein a
total psychoactive alkaloid content in the psychoactive alkaloid composition
is specified
as a result of the standardizing step. In some embodiments, the desired amount
of the
phosphorylated psychoactive alkaloid is 100% by weight of the total
phosphorylatable
psychoactive alkaloid content in the psychoactive alkaloid composition, the
process
comprising: preparing another psychoactive alkaloid composition comprising
another
psychoactive alkaloid extract of some embodiments disclosed herein wherein the
desired amount of the dephosphorylated psychoactive alkaloid is 100% by weight
of the
total phosphorylatable psychoactive alkaloid content in the other psychoactive
alkaloid
extract; mixing the psychoactive alkaloid composition and the other
psychoactive
composition in a measured ratio to obtain a psychoactive alkaloid composition
comprising the phosphorylated psychoactive alkaloid of the psychoactive
alkaloid
composition and the dephosphorylated psychoactive alkaloid of the other
psychoactive
alkaloid composition in a specific ratio; wherein the specific ratio of
phosphorylated
psychoactive alkaloid to dephosphorylated psychoactive alkaloid ranges from
1:1000 to
1000:1.
[14] A process for obtaining a psychoactive alkaloid composition with a
specific
ratio of a phosphorylated psychoactive alkaloid to a dephosphorylated
psychoactive
alkaloid, the process comprising: extracting a psychoactive alkaloid from a
dried
powdered biomass with a basified solvent to obtain a psychoactive alkaloid
liquid with a
pH greater than 10.5, wherein a majority of a total phosphorylatable
psychoactive
alkaloid content is the phosphorylated alkaloid and a remainder thereof is the
dephosphorylated alkaloid; adjusting the pH of the psychoactive alkaloid
liquid to a pH
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PSU004-CADIV1
ranging from 3.5-4.5; extracting another psychoactive alkaloid from another
dried
powdered biomass with an acidified solvent to obtain another psychoactive
alkaloid
liquid with a pH lower than 3.5, wherein all of a total phosphorylatable
psychoactive
alkaloid is the dephosphorylated alkaloid; adjusting the pH of the other
psychoactive
alkaloid liquid to a pH ranging from 3.5-4.5; evaporating a portion of the
basified solvent
from the psychoactive alkaloid liquid and a portion of the acidified solvent
from the other
psychoactive alkaloid liquid to obtain a psychoactive alkaloid extract slurry
and another
psychoactive alkaloid extract slurry respectively; mixing measured portions of
the
psychoactive alkaloid extract slurry and the other psychoactive alkaloid
extract slurry to
obtain a bulk psychoactive alkaloid extract slurry comprising the
phosphorylated
psychoactive alkaloid and the dephosphorylated psychoactive alkaloid in the
specific
ratio; standardizing the bulk psychoactive alkaloid extract slurry by adding
thereto a
measured quantity of one or more excipients to obtain a standardized bulk
slurry; and
drying the standardized bulk psychoactive alkaloid slurry to obtain the
psychoactive
alkaloid composition, wherein the phosphorylated psychoactive alkaloid and the
dephosphorylated psychoactive alkaloid are in the specific ratio; wherein the
specific
ratio of phosphorylated psychoactive alkaloid to dephosphorylated psychoactive
alkaloid ranges from 1:1000 to 1000:1. A psychoactive alkaloid composition
comprising:
a psychoactive alkaloid extract comprising a desired amount of a
phosphorylated
psychoactive alkaloid and a desired amount of a dephosphorylated psychoactive
alkaloid, wherein: the desired amount of the phosphorylated psychoactive
alkaloid is 0-
100% by weight of a total phosphorylatable psychoactive alkaloid content in
the
psychoactive alkaloid extract, and the desired amount of the dephosphorylated
psychoactive alkaloid is the remainder of the total phosphorylatable
psychoactive
alkaloid content in the psychoactive alkaloid extract; and one or more
excipients. In
some embodiments, the composition is in powder form. In some embodiments, the
desired amount of the phosphorylated psychoactive alkaloid is 0% by weight of
the total
phosphorylatable psychoactive alkaloid content in the psychoactive alkaloid
extract,
and the desired amount of the dephosphorylated psychoactive alkaloid is 100%
by
weight of the total phosphorylatable psychoactive alkaloid content in the
psychoactive
alkaloid extract. In some embodiments, the desired amount of the
phosphorylated
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PSU004-CADIV1
psychoactive alkaloid is 100% by weight of the total phosphorylatable
psychoactive
alkaloid content in the psychoactive alkaloid extract, and the desired amount
of the
dephosphorylated psychoactive alkaloid is 0% by weight of the total
phosphorylatable
psychoactive alkaloid content in the psychoactive alkaloid extract. In some
embodiments, the phosphorylated alkaloid is psilocybin, baeocystin,
norbaeocystin,
aeruginascin, or any combination selected therefrom; and the dephosphorylated
alkaloid is psilocin, norpsilocin, 4-hydroxytryptamine, N,N,N-trimethy1-4-
hydroxytryptamine, or any combination selected therefrom.
[15] A psychoactive alkaloid composition with a specific ratio of a
phosphorylated
psychoactive alkaloid and a dephosphorylated psychoactive alkaloid, the
composition
comprising: a psychoactive alkaloid extract having a total phosphorylatable
psychoactive alkaloid content of 100% of a phosphorylated psychoactive
alkaloid;
another psychoactive alkaloid extract having a total phosphorylatable
psychoactive
alkaloid content of 100% of a dephosphorylated psychoactive alkaloid; and one
or more
excipients; wherein the psychoactive alkaloid extract and the other
psychoactive
alkaloid extract are present in a proportion such that the specific ratio of
phosphorylated
psychoactive alkaloid to phosphorylated psychoactive alkaloid ranges from
1:1000 to
1000:1.
[16] The present invention is directed to an extraction process of
psychoactive
compounds from psychedelic fungus, for example, the Psilocybe cubensis species
of
psychedelic mushroom. The principal psychoactive compounds in Psilocybe
cubensis
include psilocybin and psilocin. In particular, the extraction process of
psychoactive
compounds involves drying fresh Psilocybe cubensis, followed by grinding,
extraction
with a solvent in one or more steps, one or more steps of filtration, optional
adjustment
of the pH if the solvent is acidic (acid/water/alcohol) or alkaline
(base/water/alcohol),
evaporation of the solvent, and standardization. Optionally, the process
includes drying
to result in a final powdered psilocybin mushroom extract.
[17] The invention described here consists of a never-before-described
method to
produce high-efficiency, standardized preparations of the target compounds all
while
using acceptable solvent systems.
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[18] The drying of fresh fungal biomass done in a fashion that does not
greatly
reduce the natural psilocybin, psilocin, or baeocystin concentrations.
[19] The temperature of extraction is between 5 and 95 C and uses a Ito
50:1
solvent:solid ratio of extraction. Time of extraction from 10 to 720 minutes.
Pressure
may be applied, ranging from 50kPa to 137MPa (0.5 atm to 20,000 psi). Multiple
species of psilocybe mushrooms may be used.
[20] Disclosed herein is a process for forming an extract of psychoactive
alkaloids
from psychoactive organisms such as psychedelic fungus comprising the steps
of:
soaking a biomass of dried, raw psychedelic fungus in a solvent selected from
the
group consisting of water-ethanol and water-methanol mixture in order to
dissolve the
psychoactive alkaloids in the solvent; filtering an undissolved portion of the
biomass
from the solvent; evaporating the solvent to remove the methanol or ethanol
completely, leaving a concentrated slurry or a residue that is converted to
the
concentrated slurry by adding water thereto: and standardizing the
concentrated slurry
by adding thereto a quantity of carrier measured to achieve a specified purity
of extract.
In some embodiments the solvent is water-methanol. In some embodiments the
solvent
is water-methanol, the concentration being a trace to about 100 %. In some
embodiments the solvent is water-methanol, about 10 % to about 90 %. In some
embodiments the solvent is water-methanol, about 20 % to about 80 %. In some
embodiments the solvent is water-methanol, about 30 `)/0 to about 70 %. In
some
embodiments the solvent is water-methanol, about 40 % to about 60 %. In some
embodiments the solvent is water-methanol, about 50 A to about 50 %. In some
embodiments the solvent is water-methanol, about 60 % to about 40 %. In some
embodiments the solvent is water-methanol, about 70 % to about 30 %. In some
embodiments the solvent is water-methanol, about 80 `)/0 to about 20 %. In
some
embodiments the solvent is water-methanol, about 90 % to about 10 %. In some
embodiments the solvent is water-methanol, about 100 % to a trace. In some
embodiments the solvent is water-ethanol. In some embodiments the solvent is
water-
ethanol, the concentration being a trace to about 100 %. In some embodiments
the
solvent is water-ethanol, about 10 A to about 90 %. In some embodiments the
solvent
is water-ethanol, about 20 % to about 80 %. In some embodiments the solvent is
water-
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PSU004-CADIV1
ethanol, about 30 % to about 70 %. In some embodiments the solvent is water-
ethanol,
about 40 % to about 60 %. In some embodiments the solvent is water-ethanol,
about 50
% to about 50 %. In some embodiments the solvent is water-ethanol, about 60 %
to
about 40 %. In some embodiments the solvent is water-ethanol, about 70 % to
about 30
%. In some embodiments the solvent is water-ethanol, about 80 % to about 20 %.
In
some embodiments the solvent is water-ethanol, about 90 % to about 10 %. In
some
embodiments the solvent is water-ethanol, about 100 % to a trace. In some
embodiments, the standardizing comprises measuring a psychoactive alkaloid
content
in the concentrated slurry, and using the psychoactive alkaloid content, the
specified
purity and a volume of the concentrated slurry to determine the quantity of
carrier.
[21] The present invention relates to a process for obtaining a purified
psychoactive alkaloid solution from a psychoactive alkaloid source. The
purification
process of the present invention allows for producing standardized
preparations of
psychoactive alkaloids, all while using acceptable solvent and processing
systems.
[22] Standardization is a method that can be used to solve the problem of
inconsistency in the finished product. However, when dealing with a low-
potency
feedstock material, it may be difficult to standardize the active ingredients
to a high
percentage content and achieve the desired therapeutic effects. We therefore
need to
concentrate the active ingredients beforehand, using a purification process.
It may also
be desirable to concentrate the active ingredients to a high enough degree
that the
resulting volume of the final product is limited for a specific application,
such as to fit
into a standard size two-piece capsule.
[23] A psychoactive alkaloid source is used to provide a psychoactive
alkaloid
extract. The source may be a species containing psychedelic alkaloids or a
prior extract
therefrom. Psychoactive alkaloids in the extract are adsorbed onto a resin or
other
adsorbent material, from which they are then eluted to provide a purified
psychoactive
alkaloid solution. The process may be repeated with different resins,
different pH
values, and different elution solvents. Solids present in the extract may be
removed at
various stages by filtering or centrifuging.
[24] The purification process of the present invention allows for purifying
relatively
low-potency feedstocks to result in a purified psychoactive alkaloid solution
that may
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PSU004-CADIV1
have a relatively high concentration of psychoactive alkaloid. Depending on
the
embodiment, the process may be a purification process that enriches the
psychoactive
alkaloid content of the final formulation compared to the alkaloid content in
the raw
materials. Purification may also be the removal of some of the impurities,
irrespective of
the final alkaloid content. The process of purification in the present
invention allows use
of the lowest-grade raw materials to obtain a product capable of
standardization to a
desired specification.
[25] The purification process of the present invention may be, depending on
the
embodiment, a relatively simple and robust psychoactive alkaloid purification
process,
which is suitable for the production of food-grade, nutraceutical-grade, or
pharmaceutical-grade standardized extracts, especially of psilocybin,
psilocin,
baeocystin, norbaeocystin, norpsilocin, aeruginascin, bufotenin, bufotenidine,
5-Me0-
DMT (5-methoxy-N.N-dimethyltryptamine), N,N-dimethyltryptamine (DMT), ergine
(LSA), ergonovine, ergometrine, muscimol, ibotenic acid, lysergic acid
hydroxyethylamide (LSH), elymoclavine, ergometrinine, and/or chanoclavine.
[26] The present invention also relates to a standardization process for
preparation
of standardized extracts of psychoactive alkaloids. The standardization
process of the
present invention allows for standardizing the purified psychoactive alkaloid
solution to
result in a purified psychoactive alkaloid extract with a specific
concentration of
psychoactive alkaloids. The standardization process of the present invention
may also
be a simple and cost-efficient process.
[27] The standardized psychoactive alkaloid extracts of the present
invention can
be used in, for example, medical research on the use of psychedelic substances
as
treatments for addiction, post-traumatic stress disorder, depression, cluster
headaches,
and other illnesses. They may also be used in traditional entheogenic
practices or
consumed recreationally where such activity is permitted by law.
[28] Disclosed herein is a process for obtaining a purified psychoactive
alkaloid
solution, the process comprising: extracting a psychoactive alkaloid from a
psychoactive alkaloid source to obtain a psychoactive alkaloid extract;
treating the
psychoactive alkaloid extract with an adsorbent material to obtain an adsorbed
psychoactive alkaloid; and eluting the adsorbed psychoactive alkaloid using a
solvent to
Date Recue/Date Received 2022-07-25
PSU004-CADIV1
obtain a purified psychoactive alkaloid solution, wherein the solvent is
water, an organic
solvent or a combination thereof, under basic, acidic or neutral pH.
[29] In some embodiments, the process includes: evaporating a portion of
solvent
from the purified psychoactive alkaloid solution to obtain a concentrated
slurry;
standardizing the concentrated slurry by adding thereto a quantity of
excipient
measured to provide a specific concentration of psychoactive alkaloid when the
concentrated slurry is dried; and drying the slurry by evaporating the
remaining portion
of the solvent to obtain a standardized, purified, powdered psychoactive
alkaloid extract
having the specific concentration of psychoactive alkaloid.
[30] An aim of the invention is to standardize the amount of psychoactive
alkaloid
present in the composition. Depending on the embodiment and the specific types
of
excipient added, a secondary, optional aspect of the invention is the
provision of a
psychoactive alkaloid extract in the form of a dry, flowable and shelf-stable
powder. The
powder may be used as a dietary supplement or medicine and can be added to
various
edible products, tablets, or capsules, or it may be used for medical research,
including
the study of the treatment of mental illnesses.
[31] By increasing the active alkaloid concentration through extraction and
then
titrating back to a lower, standardized alkaloid concentration, the product
achieves
consistency in bioactive content from lot to lot. By the addition of specific
types of
excipient, flowability and stability may also be improved in the composition,
as
compared to the extract.
[32] A useful formulation needs to contain a minimum amount of the active
ingredient and also be of an acceptable total size. For example, a psilocybin
dose might
be 25 mg. If this is required in a single capsule and the powder has a
concentration of
the active ingredient of only 1%, then 2500 mg of powder would be needed. This
would
be too much for a single capsule. However, if the extract can be concentrated
to
approx. 15%, then there is room to add an excipient to get it down to say, a
repeatable
10%, which now means that only 250 mg of powder is needed in the capsule,
which is
an acceptable size.
[33] While the concentration of psychoactive alkaloid in the composition
may, in
some embodiments, be lower than that found in some of the source raw material
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(mushrooms or seeds for example), it is a known concentration, which can be
stable
from batch to batch, eliminating the variability found in the natural sources.
This allows
for control and standardization of the dose, even if it is lower than some of
the raw
materials themselves.
[34] Disclosed herein is a psychoactive alkaloid composition consisting
essentially
of, by weight: 0.1-99.9% of a psychoactive alkaloid extract; one or more
preservatives
up to 10%, a flow agent up to 2%, or both the one or more preservatives up to
10% and
the flow agent up to 2%; and 0-94% of a carrier.
[35] The inventors have realized that there are occasions where it would be
beneficial to control, either by halting or promoting, the conversion of
phosphorylated
alkaloids such as psilocybin during extraction and any subsequent purification
process.
For example, there may be occasions where extracts with phosphorylated
psychoactive
alkaloids as the only or majority of the total psychoactive alkaloid in the
extract are
required. Likewise, there may be occasions where extracts with
dephosphorylated
psychoactive alkaloids as the only or majority of the total psychoactive
alkaloid in the
extract are required.
[36] For example, the alkaloid in the psychoactive extract may be entirely
psilocin,
resulting from promotion of the conversion, or all or mostly psilocybin, from
the halting
or inhibition of the conversion. There will likely, but not necessarily, be
some psilocin
present in the process where the conversion is halted, as the harvesting of
the
mushrooms can often cause unavoidable conversion into psilocin.
[37] Controlling the promotion or inhibition of dephosphorylation of the
aforementioned alkaloids to result in a psychoactive alkaloid extract with a
specific
desired content of both phosphorylated and dephosphorylated psychoactive
alkaloid
has not been seen in the industry or academia to date. Likewise, compositions
resulting
from such control have not yet been seen.
[38] Thus, there is a need in the art of a process for controlling the
dephosphorylation of the aforementioned alkaloids to result in a psychoactive
alkaloid
extract with specific desired amounts of phosphorylated and dephosphorylated
psychoactive alkaloid. Also required in the art is a psychoactive alkaloid
composition
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PSU004-CADIV1
having an accurate psychoactive alkaloid content, with specific desired
amounts of
phosphorylated and dephosphorylated psychoactive alkaloid.
[39] Disclosed herein is a process for obtaining a psychoactive alkaloid
extract
with a desired amount of a phosphorylated psychoactive alkaloid and a desired
amount
of a dephosphorylated psychoactive alkaloid, the process comprising: drying
and
pulverizing a psychoactive alkaloid source to obtain a dried powdered biomass;
extracting a psychoactive alkaloid from the dried powdered biomass with an
acidified
solvent or a basified solvent to obtain a psychoactive alkaloid liquid with a
specific pH,
wherein the specific pH is lower than 3.5 or greater than 10.5; adjusting the
pH of the
psychoactive alkaloid liquid to a pH ranging from 3.5-4.5; and evaporating the
solvent
from the psychoactive alkaloid liquid to obtain the psychoactive alkaloid
extract with the
desired amount of the phosphorylated psychoactive alkaloid and the desired
amount of
the dephosphorylated psychoactive alkaloid; wherein: the desired amount of the
phosphorylated psychoactive alkaloid is 0-100% by weight of a total
phosphorylatable
psychoactive alkaloid content in the psychoactive alkaloid extract; and the
desired
amount of the dephosphorylated psychoactive alkaloid is the remainder of the
total
phosphorylatable psychoactive alkaloid content in the psychoactive alkaloid
extract.
[40] Also disclosed is a process for obtaining a psychoactive alkaloid
composition
with a specific ratio of a phosphorylated psychoactive alkaloid to a
dephosphorylated
psychoactive alkaloid, the process comprising: extracting a psychoactive
alkaloid from a
dried powdered biomass with a basified solvent to obtain a psychoactive
alkaloid liquid
with a pH greater than 10.5, wherein a majority of a total phosphorylatable
psychoactive
alkaloid content is the phosphorylated alkaloid and a remainder thereof is the
dephosphorylated alkaloid; adjusting the pH of the psychoactive alkaloid
liquid to a pH
ranging from 3.5-4.5; extracting another psychoactive alkaloid from another
dried
powdered biomass with an acidified solvent to obtain another psychoactive
alkaloid
liquid with a pH lower than 3.5, wherein all of a total phosphorylatable
psychoactive
alkaloid is the dephosphorylated alkaloid; adjusting the pH of the other
psychoactive
alkaloid liquid to a pH ranging from 3.5-4.5; evaporating a portion of the
basified solvent
from the psychoactive alkaloid liquid and a portion of the acidified solvent
from the other
psychoactive alkaloid liquid to obtain a psychoactive alkaloid extract slurry
and another
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PSU004-CADIV1
psychoactive alkaloid extract slurry respectively; mixing measured portions of
the
psychoactive alkaloid extract slurry and the other psychoactive alkaloid
extract slurry to
obtain a bulk psychoactive alkaloid extract slurry comprising the
phosphorylated
psychoactive alkaloid and the dephosphorylated psychoactive alkaloid in the
specific
ratio; standardizing the bulk psychoactive alkaloid extract slurry by adding
thereto a
measured quantity of one or more excipients to obtain a standardized bulk
slurry; and
drying the standardized bulk psychoactive alkaloid slurry to obtain the
psychoactive
alkaloid composition, wherein the phosphorylated psychoactive alkaloid and the
dephosphorylated psychoactive alkaloid are in the specific ratio; wherein the
specific
ratio of phosphorylated psychoactive alkaloid to dephosphorylated psychoactive
alkaloid ranges from 1:1000 to 1000:1.
[41] Further disclosed is a psychoactive alkaloid composition comprising: a
psychoactive alkaloid extract comprising a desired amount of a phosphorylated
psychoactive alkaloid and a desired amount of a dephosphorylated psychoactive
alkaloid, wherein: the desired amount of the phosphorylated psychoactive
alkaloid is 0-
100% by weight of a total phosphorylatable psychoactive alkaloid content in
the
psychoactive alkaloid extract, and the desired amount of the dephosphorylated
psychoactive alkaloid is the remainder of the total phosphorylatable
psychoactive
alkaloid content in the psychoactive alkaloid extract; and one or more
excipients.
[42] Still further disclosed is a psychoactive alkaloid composition with a
specific
ratio of a phosphorylated psychoactive alkaloid and a dephosphorylated
psychoactive
alkaloid, the composition comprising: a psychoactive alkaloid extract having a
total
phosphorylatable psychoactive alkaloid content of 100% of a phosphorylated
psychoactive alkaloid; another psychoactive alkaloid extract having a total
phosphorylatable psychoactive alkaloid content of 100% of a dephosphorylated
psychoactive alkaloid; and one or more excipients; wherein the psychoactive
alkaloid
extract and the other psychoactive alkaloid extract are present in a
proportion such that
the specific ratio of phosphorylated psychoactive alkaloid to phosphorylated
psychoactive alkaloid ranges from 1:1000 to 1000:1.
[43] A method for generating a psychoactive alkaloid extract comprising
0.1% to
99% by weight of a psychoactive alkaloid from a psychoactive organism, the
method
24
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PSU004-CADIV1
comprising: providing a biomass of the psychoactive organism; contacting the
biomass
with 10 to 100 milliliters (mL) of solvent per gram (g) of the biomass; and
evaporating
the solvent from the biomass to yield the psychoactive alkaloid extract
comprising 0.1%
to 99% by weight of the psychoactive alkaloid. In some embodiments, following
(b), the
biomass is contacted with a second solvent. In some embodiments, the solvent
is
selected from 100% methanol, an alcohol:water mixture wherein the alcohol
comprises
60% to 99% of the alcohol:water mixture, an alcohol:acid mixture wherein the
alcohol
comprises 60% to 99% of the alcohol:acid mixture, an alcohol:base mixture
wherein the
alcohol comprises 60% to 99% of the alcohol:base mixture, an alcohol:water
mixture
wherein the alcohol comprises 70% to 80% of the alcohol:water mixture, an
alcohol:acid
mixture wherein the alcohol comprises 70% to 80% of the alcohol:acid mixture,
an
alcohol:base mixture wherein the alcohol comprises 70% to 80% of the
alcohol:base
mixture, acidified water, and basified water. In some embodiments, the alcohol
of the
alcohol:water mixture, the alcohol:acid mixture, or alcohol:base mixture, is a
C1-C4
primary aliphatic alcohol. In some embodiments, the C1-C4 primary aliphatic
alcohol is
ethanol or methanol. In some embodiments, the solvent is buffered to a pH of
either 4
or less, or 9 or greater. In some embodiments, the solvent has a pH of 10 or
greater
and the psychoactive alkaloid extract comprises greater than 50% of a
phosphorylated
psychoactive alkaloid. In some embodiments, the solvent has a pH of 4 or less
and the
psychoactive alkaloid extract comprises greater than 50% of a dephosphorylated
psychoactive alkaloid. In some embodiments, the solvent is buffered with
ammonium
bicarbonate, ammonium carbonate, ammonium hydroxide, calcium acetate, calcium
carbonate, calcium chloride, calcium hydroxide, calcium lactate, calcium
oxide, calcium
phosphate, dibasic, calcium phosphate monobasic, magnesium carbonate,
potassium
aluminum sulphate, potassium bicarbonate, potassium carbonate, potassium
hydroxide,
potassium lactate, potassium phosphate, dibasic, potassium pyrophosphate,
tetrabasic,
potassium phosphate tribasic, potassium tripolyphosphate, sodium acetate,
sodium
acid pyrophosphate, sodium aluminum phosphate, sodium aluminum sulphate,
sodium
bicarbonate, sodium bisulphate, sodium carbonate, sodium hexametaphosphate,
sodium hydroxide, sodium lactate, sodium phosphate dibasic, sodium phosphate
monobasic, sodium phosphate tribasic, and any combination thereof. In some
Date Recue/Date Received 2022-07-25
PSU004-CADIV1
embodiments, the solvent is buffered with acetic acid, adipic acid, ascorbic
acid,
phosphoric acid, ammonium aluminum sulphate, ammonium citrate dibasic,
ammonium
citrate monobasic, calcium citrate, calcium fumarate, calcium gluconate,
calcium
phosphate dibasic, calcium phosphate, hydrochloric acid, sulphuric acid
monobasic,
calcium phosphate tribasic, citric acid, fumaric acid, gluconic acid,
magnesium
fumarate, malic acid, phosphoric acid, potassium acid tartrate, potassium
citrate,
potassium fumarate, sodium citrate, sodium fumarate, sodium gluconate, sodium
lactate, sodium potassium hexametaphosphate, sodium potassium tartrate, sodium
potassium tripolyphosphate, sodium pyrophosphate tetrabasic, sodium
tripolyphosphate, tartaric acid, and any combination thereof. In some
embodiments, the
psychoactive organism is Psilocybe cyanescens, Psilocybe cubensis, Amanita
muscaria, or any selection thereof. In some embodiments, the psychoactive
alkaloid is
psilocybin, psilocin, baeocystin, norbaeocystin, norpsilocin, aeruginascin,
bufotenin,
bufotenidine, 5-Me0-DMT (5-methoxy-N,N-dimethyltryptamine), N,N-
dimethyltryptamine (DMT), 4-hydroxytryptamine, N,N,N-trimethyl-4-
hydroxytryptamine
ergine (LSA), ergonovine, ergometrine, muscimol, ibotenic acid, lysergic acid
hydroxyethylamide (LSH), elymoclavine, ergometrinine, and/or chanoclavine, or
any
combination thereof.
[44] A method for generating a phosphorylated psychoactive alkaloid extract
comprising 0.1`)/0 to 99% by weight of a psychoactive alkaloid from a
psychoactive
organism, the method comprising: providing a biomass of the psychoactive
organism;
contacting the biomass with 10 to 100 milliliters (mL) of solvent per gram (g)
of the
biomass wherein the solvent has a pH of 9 or greater and the psychoactive
alkaloid
comprises greater than 50% of a phosphorylated psychoactive alkaloid; and
evaporating the solvent from the biomass to yield the phosphorylated
psychoactive
alkaloid extract comprising 0.1% to 99% by weight of the psychoactive
alkaloid. In
some embodiments, following (b), the biomass is contacted with a second
solvent. In
some embodiments, the solvent is selected from 100% methanol, an alcohol:water
mixture wherein the alcohol comprises 60% to 99% of the alcohol:water mixture,
an
alcohol:acid mixture wherein the alcohol comprises 60% to 99% of the
alcohol:acid
mixture, an alcohol:base mixture wherein the alcohol comprises 60% to 99% of
the
26
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PSU004-CADIV1
alcohol:base mixture, an alcohol:water mixture wherein the alcohol comprises
70% to
80% of the alcohol:water mixture, an alcohol:acid mixture wherein the alcohol
comprises 70% to 80% of the alcohol:acid mixture, an alcohol:base mixture
wherein the
alcohol comprises 70% to 80% of the alcohol:base mixture, acidified water, and
basified
water. In some embodiments, the alcohol of the alcohol:water mixture, the
alcohol:acid
mixture, or the alcohol:base mixture, is a Cl-C4 primary aliphatic alcohol. In
some
embodiments, the C1-C4 primary aliphatic alcohol is ethanol or methanol. In
some
embodiments, the solvent is buffered with ammonium bicarbonate, ammonium
carbonate, ammonium hydroxide, calcium acetate, calcium carbonate, calcium
chloride,
calcium hydroxide, calcium lactate, calcium oxide, calcium phosphate, dibasic,
calcium
phosphate monobasic, magnesium carbonate, potassium aluminum sulphate,
potassium bicarbonate, potassium carbonate, potassium hydroxide, potassium
lactate,
potassium phosphate, dibasic, potassium pyrophosphate, tetrabasic, potassium
phosphate tribasic, potassium tripolyphosphate, sodium acetate, sodium acid
pyrophosphate, sodium aluminum phosphate, sodium aluminum sulphate, sodium
bicarbonate, sodium bisulphate, sodium carbonate, sodium hexametaphosphate,
sodium hydroxide, sodium lactate, sodium phosphate dibasic, sodium phosphate
monobasic, sodium phosphate tribasic, and any combination thereof. In some
embodiments, the solvent is buffered with acetic acid, adipic acid, ascorbic
acid,
phosphoric acid, ammonium aluminum sulphate, ammonium citrate dibasic,
ammonium
citrate monobasic, calcium citrate, calcium fumarate, calcium gluconate,
calcium
phosphate dibasic, calcium phosphate, hydrochloric acid, sulphuric acid
monobasic,
calcium phosphate tribasic, citric acid, fumaric acid, gluconic acid,
magnesium
fumarate, malic acid, phosphoric acid, potassium acid tartrate, potassium
citrate,
potassium fumarate, sodium citrate, sodium fumarate, sodium gluconate, sodium
lactate, sodium potassium hexametaphosphate, sodium potassium tartrate, sodium
potassium tripolyphosphate, sodium pyrophosphate tetrabasic, sodium
tripolyphosphate, tartaric acid, and any combination thereof. In some
embodiments, the
psychoactive organism is Psilocybe cyanescens, Psilocybe cubensis, Amanita
muscaria, or any selection thereof. In some embodiments, the phosphorylated
27
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PSU004-CADIV1
psychoactive alkaloid is psilocybin, baeocystin, norbaeocystin, aeruginascin,
or any
combination thereof.
[45] A method for generating a dephosphorylated psychoactive alkaloid
extract
comprising 0.1% to 99% by weight of a psychoactive alkaloid from a
psychoactive
organism, the method comprising: providing a biomass of the psychoactive
organism;
contacting the biomass with 10 to 100 milliliters (mL) of solvent per gram (g)
of the
biomass wherein the solvent has a pH of 4 or less and the psychoactive
alkaloid
comprises greater than 50% of a dephosphorylated psychoactive alkaloid; and
evaporating the solvent from the biomass to yield the dephosphorylated
psychoactive
alkaloid extract comprising 0.1% to 99% by weight of the psychoactive
alkaloid. In
some embodiments, following (b), the biomass is contacted with a second
solvent. In
some embodiments, the solvent is selected from 100% methanol, an alcohol:water
mixture wherein the alcohol comprises 60% to 99% of the alcohol:water mixture,
an
alcohol:acid mixture wherein the alcohol comprises 60% to 99% of the
alcohol:acid
mixture, an alcohol:base mixture wherein the alcohol comprises 60% to 99% of
the
alcohol:base mixture, an alcohol:water mixture wherein the alcohol comprises
70% to
80% of the alcohol:water mixture, an alcohol:acid mixture wherein the alcohol
comprises 70% to 80% of the alcohol:acid mixture, an alcohol:base mixture
wherein the
alcohol comprises 70% to 80% of the alcohol:base mixture, acidified water, and
basified
water. In some embodiments, the alcohol of the alcohol:water mixture, the
alcohol:acid
mixture, or the alcohol:base mixture, is a C1-C4 primary aliphatic alcohol. In
some
embodiments, the C1-04 primary aliphatic alcohol is ethanol or methanol. In
some
embodiments, the solvent is buffered with ammonium bicarbonate, ammonium
carbonate, ammonium hydroxide, calcium acetate, calcium carbonate, calcium
chloride,
calcium hydroxide, calcium lactate, calcium oxide, calcium phosphate, dibasic,
calcium
phosphate monobasic, magnesium carbonate, potassium aluminum sulphate,
potassium bicarbonate, potassium carbonate, potassium hydroxide, potassium
lactate,
potassium phosphate, dibasic, potassium pyrophosphate, tetrabasic, potassium
phosphate tribasic, potassium tripolyphosphate, sodium acetate, sodium acid
pyrophosphate, sodium aluminum phosphate, sodium aluminum sulphate, sodium
bicarbonate, sodium bisulphate, sodium carbonate, sodium hexametaphosphate,
28
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PSU004-CADIV1
sodium hydroxide, sodium lactate, sodium phosphate dibasic, sodium phosphate
monobasic, sodium phosphate tribasic, and any combination thereof. In some
embodiments, the solvent is buffered with acetic acid, adipic acid, ascorbic
acid,
phosphoric acid, ammonium aluminum sulphate, ammonium citrate dibasic,
ammonium
citrate monobasic, calcium citrate, calcium fumarate, calcium gluconate,
calcium
phosphate dibasic, calcium phosphate, hydrochloric acid, sulphuric acid
monobasic,
calcium phosphate tribasic, citric acid, fumaric acid, gluconic acid,
magnesium
fumarate, malic acid, phosphoric acid, potassium acid tartrate, potassium
citrate,
potassium fumarate, sodium citrate, sodium fumarate, sodium gluconate, sodium
lactate, sodium potassium hexametaphosphate, sodium potassium tartrate, sodium
potassium tripolyphosphate, sodium pyrophosphate tetrabasic, sodium
tripolyphosphate, tartaric acid, and any combination thereof. In some
embodiments, the
psychoactive organism is Psilocybe cyanescens, Psilocybe cubensis, Amanita
muscaria, or any selection thereof. In some embodiments, the dephosphorylated
psychoactive alkaloid is psilocin, norpsilocin, 4-hydroxytryptamine, N,N,N-
trimethy1-4-
hydroxytryptamine, or any combination thereof.
[46] This summary does not necessarily describe all features of the
invention, and
different embodiments thereof may provide at least one but not necessarily all
of the
benefits described herein.
BRIEF DESCRIPTION OF DRAWINGS
[47] The following drawings illustrate embodiments of the invention, which
should
not be construed as restricting the scope of the invention in any way.
[48] FIG. 1 is a high-level flowchart showing the key steps of a process
for
extracting psychoactive alkaloids from psychedelic fungus, according to an
embodiment
of the present invention.
[49] FIG. 2 is a flowchart showing more detailed steps of a process for
extracting
psychoactive alkaloids from Psilocybe cubensis using a 75% ethanol solvent,
according
to an embodiment of the present invention.
29
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PSU004-CADIV1
[50] FIG. 3 is a flowchart showing more detailed steps of a process for
extracting
psychoactive alkaloids from Psilocybe cubensis using a hydro-ethanol solvent,
according to an embodiment of the present invention.
[51] FIG. 4 is a flowchart showing more detailed steps of a process for
extracting
psychoactive alkaloids from Psilocybe cubensis using a water solvent,
according to an
embodiment of the present invention.
[52] FIG. 5 is a flowchart showing more detailed steps of a process for
extracting
psychoactive alkaloids from Psilocybe cyanescens using a methanol solvent,
according
to an embodiment of the present invention.
[53] FIG. 6 is a flowchart showing more detailed steps of a process for
extracting
psychoactive alkaloids from Psilocybe cubensis using a buffered acidic
solvent,
according to an embodiment of the present invention.
[54] FIG. 7 is a flowchart showing more detailed steps of a process for
extracting
psychoactive alkaloids Psilocybe cubensis using a buffered alkaline solvent,
according
to an embodiment of the present invention.
[55] FIG. 8 is a schematic diagram of the apparatus used for the extraction
of
psychoactive compounds according to an embodiment of the present invention.
[56] FIG. 9 illustrates the steps of a basic process for obtaining a
purified
psychoactive alkaloid solution, according to an embodiment of the present
invention.
[57] FIG. 10 illustrates in detail the basic and optional steps of a
process for
purification of a psychoactive alkaloid extract, according to an embodiment of
the
present invention.
[58] FIG. 11 illustrates a process for standardizing a purified
psychoactive alkaloid
solution to obtain a standardized psychoactive alkaloid extract, according to
an
embodiment of the present invention.
[59] FIG. 12 illustrates detailed steps of a process for extracting
psychoactive
alkaloids from Psilocybe cubensis, according to an embodiment of the present
invention.
[60] FIG. 13 is a schematic diagram of an apparatus used for obtaining a
purified
psychoactive alkaloid solution and standardizing the same to result in a
standardized
psychoactive alkaloid extract, according to an embodiment of the present
invention.
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PSU004-CADIV1
[61] FIG. 14 illustrates steps of a process for obtaining a standardized
psychoactive alkaloid composition, according to an embodiment of the present
invention.
[62] FIG. 15 illustrates a process for extracting psychoactive alkaloid
from
Psilocybe cubensis, according to an embodiment of the present invention.
[63] FIG. 16 illustrates detailed steps of a process for purifying
psychoactive
alkaloid from Psilocybe cubensis, according to an embodiment of the present
invention.
[64] FIG. 17 is a schematic diagram of the apparatus used for obtaining a
psychoactive alkaloid composition, according to an embodiment of the present
invention.
[65] FIG. 18 illustrates the key steps of a process for obtaining a
psychoactive
alkaloid extract with dephosphorylation control, according to an embodiment of
the
present invention.
[66] FIG. 19 illustrates in detailed steps of a process for obtaining a
psychoactive
alkaloid extract with dephosphorylation control, according to an embodiment of
the
present invention.
[67] FIG. 20 illustrates a process for standardizing a psychoactive
alkaloid extract
to obtain a standardized psychoactive alkaloid extract.
[68] FIG. 21 illustrates a process for obtaining a psychoactive alkaloid
composition
with a specific ratio of a phosphorylated psychoactive alkaloid to a
dephosphorylated
psychoactive alkaloid, according to an embodiment of the present invention.
[69] FIG. 22 illustrates detailed steps of a process for obtaining a
psychoactive
alkaloid composition with a specific ratio of a phosphorylated psychoactive
alkaloid to a
dephosphorylated psychoactive alkaloid, according to another embodiment of the
present invention.
[70] FIG. 23 illustrates a schematic diagram of the apparatus used for
obtaining a
psychoactive alkaloid extract and standardizing the same to result in a
standardized
psychoactive alkaloid extract, according to an embodiment of the present
invention.
[71] FIG. 24 is a chart demonstrating a relationship between solvent to
solid ratio
and % recovery of alkaloids.
31
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PSU004-CADIV1
[72] FIG. 25 is a chart demonstrating a relationship between time and total
alkaloid
recovery ( /0).
[73] FIG. 26 is a chart demonstrating a relationship between mass of total
alkaloids applied (mg) and recovery total alkaloids (%).
[74] FIG. 27 is a chart demonstrating a relationship between bed volumes of
eluent
(BV) and recovery (%).
[75] FIG. 28 is a chart demonstrating psilocybin content in sample PYEX-FP-
200820 over 9 months at 25 C and 65% RH
[76] FIG. 29 is a chart demonstrating a relationship between solvent:solid
ratio (mL
of solvent / g of dry mushroom) and `)/0 recovery of alkaloids (psilocin).
[77] FIG. 30 is a chart demonstrating a relationship between mass of
psilocin
applied (mg) and recovery of psilocin (%) from XAD4 adsorption breakthrough
experiments.
[78] FIG. 31 is a chart demonstrating a relationship between time (months)
and
recovery (`)/0) from PIEX long-term stability measurement.
[79] FIG. 32A is a reaction scheme of dephosphorylation of psilocybin by
Psilocybe phosphatase followed by Psilocybe laccase.
[80] FIG. 32B is a chart demonstrating a relationship between solvent
composition
(v/V/0 solvent) and % recovery of alkaloids for the effect of aqueous
extraction solvent
composition.
[81] FIG. 32C is a chart demonstrating a relationship between solvent
composition
(v/V/0 solvent) and dry mass recovery (/0) in aqueous solvent composition.
[82] FIG. 32D is a chart demonstrating a relationship between pH and
alkaloid
yield (%) for the effect of pH on alkaloid extraction yield.
[83] FIG. 32E is a chart demonstrating a relationship between extraction
solvent
concentration (methanol v/v%) and alkaloid yield CYO for the effect of solvent
composition on alkaloid yield.
DETAILED DESCRIPTION
[84] Naturally-occurring psychoactive alkaloid comprising species have
inconsistent and often low contents of active psychoactive alkaloid (e.g., 0.1-
1% dry
32
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PSU004-CADIV1
wt). Considering fresh weight, this would mean a further 20X reduction in
content due
to the large moisture content of, for example, fresh mushrooms. The content of
psychoactive alkaloid in natural sources depends on various factors such as
the type of
source, harvesting season, and type of extraction process, to name a few.
Thus, the
lack of compositions having a specific desired psychoactive alkaloid content
with no or
minimal variability between different batches is a major issue.
[85] Maintaining physical and chemical stability is another challenge with
psychoactive alkaloid compositions. Usually a psychoactive alkaloid extract is
in the
form of a sticky tar, which would be difficult to handle and formulate into
standardized
compositions with specified amounts of ingredients. Extracts are not usually
amenable
to processing due to poor flowability. Extracts themselves are often not
amenable to
drying because many of the components that are pulled out of a plant or fungus
with a
lower alcohol solvent are not "dry" at room temperature (reduced sugars, oils,
and
waxes, for example). These same compounds even in low concentration can cause
the
product to be hygroscopic and become clumpy, which makes encapsulation
impossible,
and makes tabulation difficult because the powder will not "flow" in the
equipment.
[86] Thus, exposure to moisture causes psychoactive alkaloid extracts to
absorb
moisture to form clumps and become susceptible to microbial growth. Further,
the
alkaloids in these extracts can degrade, usually because of oxidation.
[87] Psychoactive alkaloids present in natural sources can be broadly
divided into
two categories, which are phosphorylated psychoactive alkaloids and
dephosphorylated
psychoactive alkaloids, although other non-phosphorylatable psychoactive
alkaloids
may also be present.
[88] Phosphorylated psychoactive alkaloids are phosphoric acid esters of
dephosphorylated psychoactive alkaloids. For example, psilocybin is a
phosphoric acid
ester of psilocin, at the 4th position. Phosphorylated psychoactive alkaloids
are
biosynthesized in natural sources. Dephosphorylated psychoactive alkaloids are
the
bioactive forms that are converted from phosphorylated alkaloids, through
phosphatase
action or chemical hydrolysis, and released when the natural source is
damaged,
harvested, or eaten. Because of this phenomenon, phosphorylated psychoactive
alkaloids are often either partially or entirely converted to dephosphorylated
33
Date Recue/Date Received 2022-07-25
PSU004-CADIV1
psychoactive alkaloids during the alkaloid extraction process, which involves
harvesting
as a necessary prior step.
[89] Although the dephosphorylated psychoactive alkaloids are the bioactive
form
of their counterpart phosphorylated psychoactive alkaloids, dephosphorylated
psychoactive alkaloids are easily degraded into non-bioactive compounds in the
presence of light, heat, and oxygen. For example, oxidation of psilocin, the
dephosphorylated counterpart to psilocybin, a phosphorylated alkaloid produced
by
biological synthesis in mushrooms, begins rapidly when exposed to air,
especially in
solution, and heat increases the oxidation rate. From our own data, the
oxidation of
psilocin in a moist and/or high light environment begins immediately, leading
to about
10% decay within 30 minutes, 25% after 5 hours, and 40-60% at 20 hours when
shielded from light. Due to this instability of the dephosphorylated
psychoactive
alkaloids, the bioactivity of the psychoactive alkaloid extracts may also be
unstable over
time.
[90] Extracts or compositions with an active ingredient made from natural
sources
generally have increased consumer acceptance and lower cost of production
compared
to synthetic compositions. There may be potential benefits of multiple natural
compounds working synergistically, colloquially known as the "entourage" or
"halo"
effect. However, the availability of psychoactive alkaloid compositions with a
desired
specific psychoactive alkaloid content is a major challenge faced by
researchers. The
variability in the content of the psychoactive alkaloids extracted from their
natural
sources is a hurdle in trying to avoid variability in the psychoactive
alkaloid
concentration in extracted compositions. It is even more challenging to
produce
consistent formulations when the concentration of active ingredients being
extracted is
typically very low in the natural source. Maintaining physical and chemical
stability is
also an issue with these compositions. Extracts or compositions containing
psychoactive alkaloids are often not amenable to drying, processing (due to
poor
flowability), or packaging methods such as tabulation or encapsulation.
[91] This application relates to the extraction of active ingredients from
fungus and
processes of purifying them. More specifically, it relates to extracting
psychoactive
compounds from fungus and forming an extract of specific purities appropriate
for uses
34
Date Recue/Date Received 2022-07-25
PSU004-CADIV1
of the psychoactive compounds in therapeutic formulations. More specifically,
the
present invention relates to a purification process for obtaining a purified
psychoactive
alkaloid solution from an extract. Further, the present invention also relates
to a process
of forming a standardized extract from the purified psychoactive alkaloid
solution,
wherein the extract has a desired, specific concentration of psychoactive
alkaloids.
[92] This application relates to a process of obtaining psychoactive
alkaloid
extracts. More specifically, the present invention relates to controlling
dephosphorylation during extraction. Further, the present invention relates to
psychoactive alkaloid compositions with controlled dephosphorylation.
[93] This application relates to a composition. Particularly, this
application relates
to psychoactive alkaloid compositions comprising a natural extract.
A. Glossary
[94] To facilitate the understanding of this invention, a number of terms
are defined
below. Terms used herein have meanings as commonly understood by a person of
ordinary skill in the areas relevant to the present invention, unless
otherwise defined.
Terms such as "a", "an" and "the" are not intended to refer to only a singular
entity but
include the general class of which a specific example may be used for
illustration. The
terminology herein is used to describe specific embodiments of the invention,
but its
usage does not delimit the invention, except as outlined in the claims.
[95] Psilocybin fungi or psilocybin mushrooms - these are a group of fungi
that
contain at least one psychoactive alkaloid, and generally contain psilocybin
and
psilocin. They may also contain other psychoactive alkaloids such as
baeocystin,
norbaeocystin, ibotenic acid and norpsilocin. The genera of these mushrooms
include
Cope/and/a, Gymnopilus, Inocybe, Panaeolus, Pholiotina, Pluteus, Amanita, and
Psilocybe.
[96] Psilocybe mushrooms - these form a genus of gilled mushrooms in the
family
Hymenogastraceae. Most species contain the psychedelic alkaloids psilocybin,
psilocin,
and baeocystin.
[97] Psilocybin ¨ this is a psychedelic prodrug produced by numerous
species of
mushrooms, collectively known as psilocybin mushrooms. Psilocybin is converted
by
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PSU004-CADIV1
the body to psilocin, which has mind-altering effects such as euphoria and
hallucinations, but can also lead to nausea and panic attacks.
[98] The term "psychoactive alkaloid extract" or "extract" refers to a
psychoactive
alkaloid extract that is obtained after a psychoactive alkaloid source has
been extracted
according to a process described herein. The extract may be a fluid, as either
a liquid or
a slurry, or is made into a fluid by the addition of a solvent. The term
"extract" may also
be used for the dried form of the fluid extract.
[99] The term "psychoactive alkaloid extract" used herein refers to a
psychoactive
alkaloid extract obtained by an extraction process of the present invention.
The extract
can be in a solid, solid-powdered, semi-solid or a slurry form.
[100] The term "psychoactive alkaloid" as used herein refers to alkaloids
that upon
ingestion are capable of changing brain function, resulting in alterations in
perception,
mood, consciousness, cognition, or behavior, for example. Psychoactive
alkaloids are
abundant in nature and can be obtained from sources such as a fungus, an
animal, a
mycelium, a spore, a plant, a bacterium, or a yeast. Examples of psychoactive
alkaloids
include, but are not limited to, psilocybin, psilocin, baeocystin,
norbaeocystin,
norpsilocin, aeruginascin, bufotenin, bufotenidine, 5-Me0-DMT (5-methoxy-N,N-
dimethyltryptamine), N,N-dimethyltryptamine (DMT), ergine (LSA), ergonovine,
ergometrine, ibotenic acid, muscimol, lysergic acid hydroxyethylamide (LSH),
elymoclavine, ergometrinine, and/or chanoclavine. The source of the
psychoactive
alkaloid can also be an extract or a solution comprising a psychoactive
alkaloid.
[101] The term "psychoactive alkaloid" used herein refers to alkaloids that
upon
introduction to the human body are capable of changing brain function, for
example
resulting in alterations in perception, mood, consciousness, cognition, or
behavior. The
psychoactive alkaloid to which the present invention applies is either a
phosphorylated
psychoactive alkaloid or a dephosphorylated psychoactive alkaloid, and there
may be
multiple different compounds in each.
[102] The term "psychoactive alkaloid source" used herein refers to a
fungus, a
mycelium, a spore, a plant, a bacterium, a Protista, an animal or a yeast,
which has in it
a phosphorylated psychoactive alkaloid, a dephosphorylated psychoactive
alkaloid, or a
combination or both. The source of the psychoactive alkaloid can also be
another
36
Date Recue/Date Received 2022-07-25
PSU004-CADIV1
extract or a solution with a phosphorylated psychoactive alkaloid, a
dephosphorylated
psychoactive alkaloid, or a combination of both.
[103] The term "phosphorylatable psychoactive alkaloid" refers to
psychoactive
alkaloids that have phosphorylated derivatives and includes psychoactive
alkaloids in
both their phosphorylated and dephosphorylated forms.
[104] The term "psychoactive alkaloid composition" used herein can also be
referred to as "composition" and describes a mixture of psychoactive alkaloid
and one
or more excipients. The composition can be of pharmaceutical, nutraceutical,
or
veterinarian grade.
[105] The term "psychoactive alkaloid liquid" used herein refers to
psychoactive
alkaloid obtained in liquid form after a dried powdered biomass of a
psychoactive
alkaloid source has been extracted using an acidified solvent or a basified
solvent. The
liquid form can be a solution or a slurry.
[106] The term "purified psychoactive alkaloid extract" refers to a
purified extract
that is obtained after a psychoactive alkaloid extract is treated with one or
more resins
for purification as described herein, or by other means of purifying the
concentration of
the psychoactive alkaloid concentration. This purified psychoactive alkaloid
extract is
substantially free of impurities, or contains fewer impurities compared to a
similar
psychoactive alkaloid extract that has not undergone any purification. The
purified
psychoactive alkaloid extract is a fluid, either a liquid or a slurry, or is
made into a fluid
by the addition of a solvent.
[107] The term "purified psychoactive alkaloid solution" refers to a
solution of one or
more desired psychoactive alkaloids, where the solution is free of impurities
or contains
fewer impurities compared to a similar psychoactive alkaloid solution that has
not
undergone any purification. The purified solution is obtained after a
psychoactive
alkaloid extracted from its source has been purified by the purification
process of the
present invention. The impurities that are commonly encountered while
extracting
psychoactive alkaloids from a natural source include sugars, carbohydrates,
chitin,
chitosan, fats, minerals, waxes, and/or proteins. The impurities being removed
from the
psychoactive alkaloid extract will vary depending on the source of the
psychoactive
alkaloid.
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[108] The "impurities" herein are commonly undesired, but not necessarily
harmful,
substances encountered while extracting psychoactive alkaloids from
psychoactive
organisms. Impurities may include sugars, carbohydrates, chitin, chitosan,
fats,
minerals, waxes, and/or proteins. The impurities being removed from a
psychoactive
alkaloid extract will vary depending on the source of the psychoactive
alkaloid. Their
removal increases the concentration of the desired psychoactive alkaloids
remaining in
the extract.
[109] The term "standardized psychoactive alkaloid extract" is used herein
to
describe a formulation derived from the purified psychoactive alkaloid
solution, which
has been standardized using a process described herein. The standardized
psychoactive alkaloid extract includes psychoactive alkaloids in a specific
concentration.
[110] The term "resin" as used herein is intended to refer to a solid or
highly-viscous
substance of plant, mineral, or synthetic origin that has been typically
converted into a
polymer. Resins are usually mixtures of organic compounds. They are typically
used in
chromatographic techniques as a stationary phase to purify and separate
compounds
depending on their polarity. Resins can be physically or chemically modified
to provide
specificity to bind or repel particular molecules within sometimes very
complex
mixtures. A resin is an example of an adsorbent material.
[111] As used herein, the term "ion exchange resin" refers to an insoluble
organic
polymer containing charged groups that tract and hold oppositely charged ions
present
in a surrounding solution in exchange for counterions previously held.
Suitable ion
exchange resins to be used herein contain cationic groups that tract and hold
anions
present in a surrounding solution and are sometimes referred to as "anion ion-
exchange
resins". Similarly, other ion exchange resins to be used herein contain
anionic groups
that tract and hold cations present in a surrounding solution and are
sometimes referred
to as "cation ion-exchange resins".
[112] The term "macroporous resin" as used herein refers to a nonionic,
cation, or
anion resin with very small, highly cross-linked polymer particles with tiny
channels.
Macroporous resins are generally used for the adsorption of organic
constituents due to
their hydrophobic properties and are thus used to separate and purify
compounds. The
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adsorption capacity of macroporous resins not only correlates with the
physical and
chemical properties of the adsorbent, but also with the size and chemical
features of the
adsorbed substance.
[113] The term "adsorbed psychoactive alkaloid" refers to one or more
alkaloids that
are adsorbed onto an adsorbent material such as a resin.
[114] The term "other adsorbent material" as used herein refers to
materials which
can be used in place of the resin(s) to adsorb the psychoactive alkaloids.
Examples of
such materials include, but are not limited to, zeolites, clays, bentonite,
minerals,
alumina, diatomaceous earth, activated carbon, charred biomass, and others.
[115] The term "purification process" may be used herein to refer to the
process
described herein, i.e. a process for obtaining a purified psychoactive
alkaloid solution.
The purification process is a separate process to the standardization process.
[116] The term "standardization" when used herein refers to a process for
obtaining
a standardized psychoactive alkaloid composition, i.e., a composition with a
defined
concentration by weight of psychoactive alkaloid.
[117] The term "standardization process" as used herein refers to the
process of
obtaining a psychoactive alkaloid extract that has a defined percentage
content of
psychoactive alkaloids. The standardization process may be applied to an
extract that
has gone through a purification process or to an extract that has not gone
through a
purification process.
[118] As used herein, the expression "standardizing" the psychoactive
alkaloid
slurry or bulk psychoactive alkaloid extract slurry refers to adding an
excipient to an
extract to obtain a slurry with a specific, total concentration of alkaloid,
by weight. The
slurry may then be dried to form a powdered composition with a pre-calculated
percentage concentration by weight of psychoactive alkaloid. The total amount
of
alkaloid content can be specified to an accuracy of up to three significant
figures.
[119] The term "psychoactive alkaloid composition" or "composition" or
"standardized composition" or "standardized purified composition" is used
herein to
describe a composition including a psychoactive alkaloid extract or a purified
psychoactive alkaloid extract, which has been standardized by the addition of
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excipients according to a presently described process. The standardized
psychoactive
alkaloid composition includes the psychoactive alkaloid in a specific amount.
[120] As used herein, the term "specific amount" when referring to a
psychoactive
alkaloid content means a desired percentage, accurate to one or two decimal
places or
one or two significant figures, of the psychoactive alkaloid content in a
psychoactive
alkaloid composition. The specific amount is defined as a percentage by weight
and can
be selected by a person of skill in the art according to preference.
[121] The term "excipient" means any component added to an active
ingredient to
make a composition. An excipient is inert in relation to the active
ingredient, in that it
essentially does not act in the same way as the active ingredient. An
excipient may be
completely inert, or it may have some other property that protects the
integrity of the
active ingredient or assists its uptake into the human body. There are
multiple types of
excipient, each having a different purpose, and a given excipient may fulfill
more than
one purpose. Examples of types of excipient include flowability agents,
flavorants,
colorants, palatants, antioxidants, bioavailability-increasing agents,
viscosity modifying
agents, tonicity agents, drug carriers, sustained-release agents, comfort-
enhancing
agents, emulsifiers, solubilizing aids, lubricants, binding agents and
stabilizing agents.
Specific excipients include pectin, rice husks, rice, xanthum gum, gum arabic,
beta
cyclodextrin, alpha cyclodextrin, microcrystalline cellulose, sorbitol,
dextrose, guar gum,
acacia gum, cellulose gum, talc, magnesium stearate.
[122] The phrase "one or more excipients" is used herein to refer that one
excipient
or more than one excipient can be used in any combination. The number of
excipients
to be used will be at the discretion of a person skilled in the art, and they
may be of
different types.
[123] The term "carrier" means an excipient that aids in delivery of the
active
ingredient or provides bulk to the composition. The amount of carrier included
in a
composition can vary widely in order to control the concentration of the
active ingredient
in the composition. An example of a carrier is mannitol, starch, maltodextrin,
tapioca
maltodextrin or rice maltodextrin, alpha and beta cyclodextrin,
microcrystalline cellulose
(MCC), gum arabic, xanthum gum, guar gum, or cellulose gum. In some
embodiments,
the starch is potato starch, corn starch, tapioca starch, arrowroot starch,
wheat starch,
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rice starch, sweet potato starch, sago starch, mung bean starch, and any
combination
of thereof. In some embodiments, mannitol is a cryoprotectant (allowing for
efficient
freeze-drying) and bulking agent.
[124] The term "flow agent", "flowability agent" or "anti-caking agent" or
"anti-
adherent" means an excipient that prevents or reduces the formation of lumps
in a
powdered composition. An example of a flow agent is silicon dioxide, stearic
acid,
magnesium stearate, or talc.
[125] The term "preservative" means an excipient that is added to the
composition
to prevent microbial growth or microbial degradation of the composition.
Examples of
preservative are ascorbic acid, citric acid, lactose, vitamin A, vitamin E,
retinyl
palmitate, selenium, sodium citrate, sodium ascorbate, calcium ascorbate,
sodium
benzoate, and potassium benzoate.
[126] The term "purified water" includes deionized water, distilled water,
reverse
osmosis water, or otherwise purified water which is substantially without free
ions.
[127] The term "substantially" as used herein refers to a majority of, or
mostly, as in
at least about 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9%,
99.99%, or at least about 99.999% or more.
[128] The term "specific pH" herein refers to a desired pH value of a
solvent or a
psychoactive alkaloid liquid obtained by adding an acidified solvent or a
basified
solvent.
[129] The term "specific pH psychoactive alkaloid solution" used herein
refers to a
solution that is obtained after addition of a suitable acid or a base to a
psychoactive
alkaloid extract to achieve a solution with a desired pH level.
[130] The term "% wt" is used to describe the weight percentage of one
component
in a mixture of components.
[131] The term "a trace" herein refers more than, but close to about 0
`)/0.
[132] The term "about" herein refers to 10%, 20%, 30%, 40%, or 50%, or
to
the nearest significant figure.
[133] The term "specific ratio" herein refers to a weight ratio between a
phosphorylated psychoactive alkaloid and a dephosphorylated psychoactive
alkaloid
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present in a psychoactive alkaloid composition. The ratio can be altered by a
person of
skill in the art according to preference.
[134] The term "desired amount" herein refers to an amount of a
phosphorylated
psychoactive alkaloid or a dephosphorylated psychoactive alkaloid in a total
phosphorylatable psychoactive alkaloid content, in the psychoactive alkaloid
liquid,
extract or composition. The amount of each of these alkaloids is controlled by
the
process for making the psychoactive alkaloid extract or psychoactive alkaloid
composition. The amount can be altered by a person of skill in the art
according to
preference. The amount is usually a percentage ratio by weight that may be
accurate
up to two significant figures.
[135] The term "therapeutic effects" is intended to qualify the amount of
active
ingredients required in the treatment of a disease or disorder or on the
effecting of a
clinical endpoint. Reference to "treatment" of a patient is intended to
include
prophylaxis. Treatment may also be preemptive in nature, i.e., it may include
prevention
of disease. Prevention of a disease may involve complete protection from
disease, for
example as in the case of prevention of infection with a pathogen or may
involve
prevention of disease progression. For example, prevention of a disease may
not mean
complete foreclosure of any effect related to the diseases at any level, but
instead may
mean prevention of the symptoms of a disease to a clinically significant or
detectable
level. Prevention of diseases may also mean prevention of progression of a
disease to
a later stage of the disease.
[136] In some embodiments, as the ranges become narrower and more central
compared to the greatest range, the properties of the embodiments generally
become
more balanced, such properties being solubility, viscosity, flowability,
stability, taste,
potency, immediate potency, delayed potency, cost of production, efficiency of
production, production time, compatibility of the psychoactive alkaloid
composition,
psychoactive efficacy of the psychoactive alkaloid extract, psychoactive
efficacy of the
psychoactive alkaloid composition, and so on. As the ranges become narrower
towards
one extreme or other of the widest range, a particular property of the
composition or
process becomes more pronounced relative to the other properties. The specific
range
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is to be chosen depending on how the properties are to be traded-off against
each
other.
[137] Throughout the description, specific details have been set forth in
order to
provide a more thorough understanding of the invention. However, the invention
may
be practiced without these particulars. In other instances, well-known
elements have
not been shown or described in detail and repetitions of steps and features
have been
omitted to avoid unnecessarily obscuring the invention. Accordingly, the
specification
and drawings are to be regarded in an illustrative, rather than a restrictive,
sense.
[138] It will be clear to one having skill in the art that further
variations to the specific
details disclosed herein can be made, resulting in other embodiments that are
within the
scope of the invention disclosed. Steps in the flowchart may be performed in a
different
order, other steps may be added, or one or more may be removed without
altering the
main outcome of the process.
B. Compositions
Source of Psychoactive Alkaloid Extract
[139] In one embodiment, the psychoactive alkaloid source is a psychoactive
organism. In one embodiment, the psychoactive organism is a fungus. In one
embodiment, the fungus is a mushroom from the genus Conocybe, Cope/and/a,
Galerina, Gymnopilus, Inocybe, Panaeolus, Pholiotina, Pluteus or Psilocybe, or
any
combination of mushrooms selected therefrom. In one embodiment, gills, caps,
stems,
or the whole of the fungi is used as the alkaloid source.
[140] The psychoactive organism may be a fungus, a mycelium, an animal, a
spore,
a plant, a bacterium, a protista, or a yeast. The psychoactive alkaloid source
in some
embodiments may be a prior extract of one or more psychoactive alkaloids,
where the
prior extract is to undergo a further extraction process. The psychoactive
alkaloid may
include, but is not limited to, psilocybin, psilocin, baeocystin,
norbaeocystin, norpsilocin,
aeruginascin, bufotenin, bufotenidine, 5-methoxy-N,N-dimethyltryptamine (5-Me0-
DMT), N,N-dimethyltryptamine (DMT), ergine (LSA), ergonovine, ergometrine,
muscimol, ibotenic acid, lysergic acid hydroxyethylamide (LSH), elymoclavine,
ergometrinine, and/or chanoclavine, or any combination selected therefrom. It
is
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possible that other psychoactive alkaloids, not yet discovered, may also be
extracted
using the methods disclosed herein.
[141] In some embodiments, the psychoactive alkaloid is a combination of
psilocybin and psilocin. In another embodiment, the psychoactive alkaloid is
psilocybin.
In yet another embodiment, the psychoactive alkaloid is psilocin.
[142] Although the examples of the present invention have been formulated
specifically using Psilocybe cubensis and Anadenanthera peregrina as sources
to
obtain a psychoactive alkaloid extract, the extract including psilocybin and
psilocin in
the first case and bufotenin, bufotenidine, and 5-Me0-DMT in the second, other
sources are also possible. A person skilled in the art would appreciate that
Psilocybe
cubensis and Anadenanthera peregrina can be readily substituted by other
sources of
psychoactive alkaloids to obtain a variety of purified psychoactive alkaloids
having
similar properties, such alkaloids being, besides those mentioned above,
baeocystin,
norbaeocystin, norpsilocin, aeruginascin, N,N-dimethyltryptamine (DMT), ergine
(LSA),
ibotenic acid, ergonovine, ergometrine, muscimol, lysergic acid
hydroxyethylamide
(LSH), elymoclavine, ergometrinine, and/or chanoclavine, to name a few, and to
result
in compositions with similar efficacy and efficiency as well. For example, the
venom of
the toad Incilius alvarius, the Anandenanthera colubrina tree or the Amanita
muscaria
mushroom may be used as other sources of psychoactive alkaloids. Note that the
lists
of sources and psychoactive alkaloids are included to provide examples and are
non-
exhaustive lists.
Psychoactive Ingredient
[143] In some embodiments, the present disclosure comprises a composition
having, by weight, 2-99.7 % of a psychoactive alkaloid extract, and 0.3-98
/0, i.e., the
remainder, being one or more excipients. In some embodiments, the psychoactive
ingredient is present in the composition at, by weight, about 50 % to about
99.7 %. In
some embodiments, the psychoactive ingredient is present in the composition
at, by
weight, about 50 % to about 60 %, about 50 % to about 70 %, about 50 % to
about 80
%, about 50 % to about 90 `)/0, about 50 A to about 92 %, about 50 % to about
94 %,
about 50 `)/0 to about 96 `)/0, about 50 % to about 98 %, about 50 `Yo to
about 99 %, about
50 % to about 99.7 %, about 60 % to about 70 %, about 60 % to about 80 %,
about 60
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% to about 90 %, about 60 `)/0 to about 92 `)/0, about 60 `)/0 to about 94
A), about 60 % to
about 96 %, about 60 % to about 98 %, about 60 % to about 99 %, about 60 A to
about
99.7 `)/0, about 70 % to about 80 %, about 70 % to about 90 %, about 70 % to
about 92
%, about 70 % to about 94 %, about 70 % to about 96 %, about 70 % to about 98
%,
about 70 % to about 99 %, about 70 % to about 99.7 %, about 80 A to about 90
%,
about 80 `)/0 to about 92 `)/0, about 80 % to about 94 %, about 80 `)/0 to
about 96 `)/0, about
80 % to about 98 %, about 80 % to about 99 %, about 80 % to about 99.7 %,
about 90
% to about 92 %, about 90 `)/0 to about 94 `)/0, about 90 `)/0 to about 96
`)/0, about 90 % to
about 98 %, about 90 % to about 99 %, about 90 % to about 99.7 %, about 92 %
to
about 94 /.0, about 92 A to about 96 %, about 92 % to about 98 %, about 92 %
to about
99 %, about 92 % to about 99.7 %, about 94 % to about 96 %, about 94 % to
about 98
%, about 94 % to about 99 %, about 94 % to about 99.7 %, about 96 % to about
98 %,
about 96 `)/0 to about 99 `)/0, about 96 `)/0 to about 99.7 %, about 98 % to
about 99 %,
about 98 % to about 99.7 %, or about 99 % to about 99.7 %. In some
embodiments,
the psychoactive ingredient is present in the composition at, by weight, about
50 %,
about 60 %, about 70 %, about 80 %, about 90 %, about 92 %, about 94 %, about
96
%, about 98 %, about 99 %, or about 99.7 %. In some embodiments, the
psychoactive
ingredient is present in the composition at, by weight, at least about 50 %,
about 60 %,
about 70 %, about 80 %, about 90 %, about 92 %, about 94 %, about 96 %, about
98
%, or about 99 %. In some embodiments, the psychoactive ingredient is present
in the
composition at, by weight, at most about 60 %, about 70 %, about 80 %, about
90 %,
about 92 %, about 94 %, about 96 %, about 98 %, about 99 %, or about 99.7 %.
[144] While a broad range of psychoactive extract is possible, if it is
above 90%
(e.g., as much as 99.9%), it is likely not to be dryable and/or flowable.
Nevertheless,
such a composition, when standardized, may still have its uses, due to the
reliability
and repeatability of the psychoactive alkaloid content. Below about 2%, the
composition may be considered to provide too low a dose of psychoactive
alkaloid.
However, compositions with less than 2% (e.g., down to 0.1%) may be possible
if
micro-dosing is desired. In some embodiments, the psychoactive ingredient is
present
in the composition at, by weight, about 0.05 % to about 5 %. In some
embodiments,
the psychoactive ingredient is present in the composition at, by weight, about
5 % to
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about 4 %, about 5 % to about 3 A, about 5 % to about 2 %, about 5 % to about
1 A,
about 5 % to about 0.8 %, about 5 % to about 0.6 %, about 5 % to about 0.4 A,
about 5
% to about 0.2 A, about 5 % to about 0.1 A, about 5 % to about 0.05 %, about
4 % to
about 3 %, about 4 % to about 2 A, about 4 A to about 1 %, about 4 A to
about 0.8 A,
about 4 % to about 0.6 %, about 4 % to about 0.4 %, about 4 % to about 0.2 A,
about 4
% to about 0.1 A, about 4 % to about 0.05 %, about 3 % to about 2 %, about 3
% to
about 1 %, about 3 % to about 0.8 %, about 3 % to about 0.6 %, about 3 A to
about 0.4
A, about 3 % to about 0.2 %, about 3 % to about 0.1 %, about 3 % to about 0.05
A,
about 2 % to about 1 %, about 2 % to about 0.8 A, about 2 % to about 0.6 %,
about 2
% to about 0.4 %, about 2 % to about 0.2 A, about 2 % to about 0.1 %, about 2
A to
about 0.05 A, about 1 % to about 0.8 %, about 1 % to about 0.6 A, about 1 %
to about
0.4 %, about 1 % to about 0.2 %, about 1 % to about 0.1 %, about 1 A to about
0.05 %,
about 0.8 % to about 0.6 %, about 0.8 % to about 0.4 A, about 0.8 % to about
0.2 A,
about 0.8 % to about 0.1 A, about 0.8 % to about 0.05 A, about 0.6 % to
about 0.4 %,
about 0.6 % to about 0.2 A, about 0.6 % to about 0.1 %, about 0.6 % to about
0.05 %,
about 0.4 % to about 0.2 A, about 0.4 % to about 0.1 %, about 0.4 A to about
0.05 %,
about 0.2 A to about 0.1 %, about 0.2 % to about 0.05 %, or about 0.1 A to
about 0.05
%. In some embodiments, the psychoactive ingredient is present in the
composition at,
by weight, about 5 %, about 4 %, about 3 %, about 2 %, about 1 A, about 0.8
%, about
0.6 A, about 0.4 A, about 0.2 %, about 0.1 %, or about 0.05 A. In some
embodiments,
the psychoactive ingredient is present in the composition at, by weight, at
least about 5
%, about 4 %, about 3 A, about 2 A, about 1 %, about 0.8 %, about 0.6 A,
about 0.4
%, about 0.2 A, or about 0.1 %. In some embodiments, the psychoactive
ingredient is
present in the composition at, by weight, at most about 4 %, about 3 %, about
2 A,
about 1 A, about 0.8 %, about 0.6 %, about 0.4 A, about 0.2 A, about 0.1
A, or about
0.05 %.
[145] In some embodiments, the psychoactive alkaloid extract has a
psychoactive
alkaloid concentration ranging from 0.1% to 99% by weight of the extract. It
may be in
the range of 1-10% dry wt/wt% for the non-purified extract concentration.
However, as
the composition may be made with purified extract, the psychoactive
concentration
could be as high as 99%. In some embodiments, the psychoactive alkaloid
extract has
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a psychoactive alkaloid concentration ranging from 1.03% to 75.22% by weight
of the
dry extract. In some embodiments, the psychoactive alkaloid extract has a
psychoactive alkaloid concentration ranging from 1.03% to 3.02% by weight of
the
extract. In other embodiments, the psychoactive alkaloid extract is a purified
psychoactive alkaloid extract. In some embodiments, the purified psychoactive
alkaloid
extract has a psychoactive alkaloid concentration ranging from 10% to 99% by
weight
of the extract. In other embodiments, the purified psychoactive alkaloid
extract has a
psychoactive alkaloid concentration ranging from 16.12 % to 75.22% by weight
of the
extract. In some embodiments, the psychoactive ingredient is present in the
composition at, by weight, about 5 % to about 76 %. In some embodiments, the
psychoactive ingredient is present in the composition at, by weight, about 5 %
to about
%, about 5 % to about 20 %, about 5 % to about 30 %, about 5 % to about 40 %,
about 5 % to about 50 %, about 5 % to about 60 %, about 5 % to about 70 %,
about 5
% to about 75%, about 5% to about 76 %, about 10 % to about 20 %, about 10 %
to
about 30 `)/0, about 10 % to about 40 `)/0, about 10 % to about 50 `)/0, about
10 % to about
60%, about 10% to about 70%, about 10% to about 75%, about 10% to about 76%,
about 20 `)/0 to about 30 %, about 20 % to about 40 %, about 20 % to about 50
%, about
% to about 60 %, about 20 % to about 70 %, about 20 % to about 75 %, about 20
%
to about 76 %, about 30 % to about 40 %, about 30 % to about 50 %, about 30 %
to
about 60 `)/0, about 30 % to about 70 %, about 30 % to about 75 %, about 30 %
to about
76 %, about 40 % to about 50 %, about 40 A to about 60 %, about 40 % to about
70 %,
about 40 `)/0 to about 75 /.0, about 40 A to about 76 %, about 50 % to about
60 %, about
50 % to about 70 %, about 50 % to about 75 %, about 50 % to about 76 %, about
60 %
to about 70 %, about 60 % to about 75 %, about 60 % to about 76 %, about 70 %
to
about 75 %, about 70 % to about 76 %, or about 75 % to about 76 %. In some
embodiments, the psychoactive ingredient is present in the composition at, by
weight,
about 5 %, about 10 %, about 20 %, about 30 %, about 40 %, about 50 /.0,
about 60 %,
about 70 %, about 75 %, or about 76 %. In some embodiments, the psychoactive
ingredient is present in the composition at, by weight, at least about 5 %,
about 10 `)/0,
about 20 %, about 30 %, about 40 %, about 50 %, about 60 %, about 70 %, or
about 75
%. In some embodiments, the psychoactive ingredient is present in the
composition at,
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by weight, at most about 10 %, about 20 %, about 30 %, about 40 %, about 50
A),
about 60 %, about 70 %, about 75 %, or about 76 %.
[146] The composition of the present invention has the psychoactive
alkaloid
present in a specific amount. In some embodiments, the specific amount of
psychoactive alkaloid is accurate to one significant figure. In another
embodiment, the
specific amount psychoactive alkaloid is accurate to two, three or four
significant
figures. The presence of the psychoactive alkaloid in a specific amount in the
composition allows for the same desired specific amount of the psychoactive
alkaloid to
be present in various batches of the psychoactive alkaloid composition.
[147] The composition of the present invention is in a powder form. The
components of the composition are also in powder form. The composition of the
present invention may be in the form of a free-flowing powder depending on the
embodiment. Such compositions are thus easy to handle during manufacturing and
packaging processes. Further, the dry, free-flowing powder form allows the
composition to be free from clumps and not be as susceptible to microbial
growth as a
composition with clumping due to moisture absorption.
Phosphorylatable Psychoactive Alkaloid
[148] In one embodiment, the present invention also relates to a
psychoactive
alkaloid composition. The psychoactive alkaloid composition includes a
specific
amount of a total phosphorylatable psychoactive alkaloid content, made up of a
desired
amount of a phosphorylated psychoactive alkaloid and a desired amount of a
dephosphorylated psychoactive alkaloid, and one or more excipients.
[149] In some embodiments, the phosphorylated alkaloid is psilocybin,
baeocystin,
norbaeocystin, aeruginascin, or any combination therefrom; and the
dephosphorylated
alkaloid is psilocin, norpsilocin, 4-hydroxytryptamine, N,N,N-trimethy1-4-
hydroxytryptamine, or any combination therefrom. In other embodiments, the
phosphorylated alkaloid is psilocybin and the dephosphorylated alkaloid is
psilocin.
[150] In some embodiments, any source that contains phosphorylated
psychoactive
alkaloids may be used as the psychoactive alkaloid source.
[151] In one embodiment, the specific amount of the total phosphorylatable
psychoactive alkaloid content in the psychoactive alkaloid composition ranges
from
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0.1-99% by weight of the composition. In another embodiment, the specific
amount of
the total phosphorylatable psychoactive alkaloid content in the psychoactive
alkaloid
composition ranges from 0.1-10% by weight of the composition. In an exemplary
embodiment, the specific amount of the total phosphorylatable psychoactive
alkaloid
content in the psychoactive alkaloid composition is 0.533% by weight of the
composition. In yet another exemplary embodiment, the specific amount of the
total
phosphorylatable psychoactive alkaloid content in the psychoactive alkaloid
composition is 0.501% by weight of the composition.
[152] In one embodiment, the desired amount of the phosphorylated
psychoactive
alkaloid is 0-100% by weight of a total phosphorylatable psychoactive alkaloid
content
in the psychoactive alkaloid extract, and the desired amount of the
dephosphorylated
psychoactive alkaloid is the remainder of the total phosphorylatable
psychoactive
alkaloid content in the psychoactive alkaloid extract. In one embodiment, the
desired
amount of the phosphorylated psychoactive alkaloid is 10-90% by weight of the
total
phosphorylatable psychoactive alkaloid content in the psychoactive alkaloid
extract,
and the desired amount of the dephosphorylated psychoactive alkaloid is the
remainder
of the total phosphorylatable psychoactive alkaloid content in the
psychoactive alkaloid
extract. In another embodiment, the desired amount of the phosphorylated
psychoactive alkaloid is 0% by weight of the total phosphorylatable
psychoactive
alkaloid content in the psychoactive alkaloid extract, and the desired amount
of the
dephosphorylated psychoactive alkaloid is 100% by weight of the total
phosphorylatable psychoactive alkaloid content in the psychoactive alkaloid
extract. In
yet another embodiment, the desired amount of the phosphorylated psychoactive
alkaloid is 100% by weight of the total phosphorylatable psychoactive alkaloid
content
in the psychoactive alkaloid extract, and the desired amount of the
dephosphorylated
psychoactive alkaloid is 0% by weight of the total phosphorylatable
psychoactive
alkaloid content in the psychoactive alkaloid extract.
[153] In some embodiments, a maximum desired amount of the phosphorylated
alkaloid is limited by an amount of the dephosphorylated alkaloid in the
psychoactive
alkaloid source.
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[154] In an exemplary embodiment, the desired amount of the phosphorylated
psychoactive alkaloid is 0.503% by weight of the psychoactive alkaloid
composition;
and the desired amount of the dephosphorylated psychoactive alkaloid is by
0.03%
weight of the psychoactive alkaloid composition. In yet another exemplary
embodiment, the desired amount of the phosphorylated psychoactive alkaloid is
0.00%
by weight of the psychoactive alkaloid composition, and the desired amount of
the
dephosphorylated psychoactive alkaloid is by 0.501% weight of the psychoactive
alkaloid composition.
[155] In one embodiment, the desired amount of the phosphorylated
psychoactive
alkaloid is 100% by weight of the total psychoactive alkaloid content in the
psychoactive
alkaloid extract.
[156] In one embodiment, the desired amount of the dephosphorylated
psychoactive alkaloid is 100% by weight of the total psychoactive alkaloid
content in the
psychoactive alkaloid extract.
[157] The dephosphorylated psychoactive alkaloids are quicker in becoming
bioavailable than their phosphorylated psychoactive alkaloids counterparts.
Thus, the
psychoactive compositions comprising dephosphorylated psychoactive alkaloids
100%
by weight in the total phosphorylatable psychoactive alkaloid content may
exhibit
therapeutic effects faster than psychoactive alkaloid compositions comprising
phosphorylated psychoactive alkaloids as the majority alkaloids in the total
phosphorylatable psychoactive alkaloid content.
[158] In one embodiment, the psychoactive alkaloid composition is obtained
by the
process for obtaining a psychoactive alkaloid extract with a desired amount of
a
phosphorylated psychoactive alkaloid and a desired amount of a
dephosphorylated
psychoactive alkaloid of the present invention.
[159] In one embodiment, the present invention also relates to a
psychoactive
alkaloid composition with a specific ratio of a phosphorylated psychoactive
alkaloid and
a dephosphorylated psychoactive alkaloid. The composition includes a
psychoactive
alkaloid extract having a total phosphorylatable psychoactive alkaloid content
of 100%
by weight of a phosphorylated psychoactive alkaloid, another psychoactive
alkaloid
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extract having a total phosphorylatable psychoactive alkaloid content of 100%
by
weight of a dephosphorylated psychoactive alkaloid, and one or more
excipients.
[160] In some embodiments, the composition includes a psychoactive alkaloid
extract having a total phosphorylated psychoactive alkaloid content of a trace
by weight
to about 100 % by weight. In some embodiments, the composition includes a
psychoactive alkaloid extract having a total phosphorylated psychoactive
alkaloid
content of a trace by weight to about 10 % by weight, a trace by weight to
about 20 %
by weight, a trace by weight to about 30 % by weight, a trace by weight to
about 40 %
by weight, a trace by weight to about 50 % by weight, a trace by weight to
about 60 %
by weight, a trace by weight to about 70 % by weight, a trace by weight to
about 80 %
by weight, a trace by weight to about 90 % by weight, a trace by weight to
about 95 %
by weight, a trace by weight to about 100 % by weight, about 10 % by weight to
about
20 hi by weight, about 10 % by weight to about 30 % by weight, about 10 % by
weight
to about 40 % by weight, about 10 % by weight to about 50 % by weight, about
10 % by
weight to about 60 A by weight, about 10 % by weight to about 70 A by
weight, about
% by weight to about 80 % by weight, about 10 % by weight to about 90 % by
weight, about 10 % by weight to about 95 A by weight, about 10 % by weight to
about
100 % by weight, about 20 % by weight to about 30 % by weight, about 20 % by
weight
to about 40 % by weight, about 20 % by weight to about 50 % by weight, about
20 % by
weight to about 60 A by weight, about 20 % by weight to about 70 A by
weight, about
% by weight to about 80 A by weight, about 20 % by weight to about 90 A) by
weight, about 20 A by weight to about 95 % by weight, about 20 % by weight to
about
100 % by weight, about 30 % by weight to about 40 % by weight, about 30 % by
weight
to about 50 % by weight, about 30 % by weight to about 60 % by weight, about
30 % by
weight to about 70 A by weight, about 30 % by weight to about 80 A by
weight, about
% by weight to about 90 % by weight, about 30 % by weight to about 95 A) by
weight, about 30 A by weight to about 100 % by weight, about 40 % by weight
to about
50 % by weight, about 40 % by weight to about 60 % by weight, about 40 % by
weight
to about 70 % by weight, about 40 % by weight to about 80 % by weight, about
40 A by
weight to about 90 % by weight, about 40 % by weight to about 95 A by weight,
about
% by weight to about 100 A by weight, about 50 % by weight to about 60 % by
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weight, about 50 % by weight to about 70 A) by weight, about 50 % by weight
to about
80 % by weight, about 50 % by weight to about 90 % by weight, about 50 % by
weight
to about 95 A by weight, about 50 A by weight to about 100 % by weight,
about 60 %
by weight to about 70 % by weight, about 60 % by weight to about 80 % by
weight,
about 60 % by weight to about 90 % by weight, about 60 % by weight to about 95
% by
weight, about 60 % by weight to about 100 % by weight, about 70 % by weight to
about
80 % by weight, about 70 % by weight to about 90 % by weight, about 70 % by
weight
to about 95 A) by weight, about 70 % by weight to about 100 % by weight,
about 80 %
by weight to about 90 % by weight, about 80 % by weight to about 95 % by
weight,
about 80 A by weight to about 100 % by weight, about 90 % by weight to about
95 %
by weight, about 90 % by weight to about 100 A by weight, or about 95 % by
weight to
about 100 % by weight. In some embodiments, the composition includes a
psychoactive alkaloid extract having a total phosphorylated psychoactive
alkaloid
content of a trace by weight, about 10 % by weight, about 20 A by weight,
about 30 %
by weight, about 40 /.3 by weight, about 50 A by weight, about 60 A by
weight, about
70 % by weight, about 80 % by weight, about 90 % by weight, about 95 % by
weight, or
about 100 % by weight. In some embodiments, the composition includes a
psychoactive alkaloid extract having a total phosphorylated psychoactive
alkaloid
content of at least a trace by weight, about 10 A by weight, about 20 % by
weight,
about 30 A by weight, about 40 A) by weight, about 50 % by weight, about 60
A by
weight, about 70 % by weight, about 80 % by weight, about 90 % by weight, or
about
95 A by weight. In some embodiments, the composition includes a psychoactive
alkaloid extract having a total phosphorylated psychoactive alkaloid content
of at most
about 10 % by weight, about 20 % by weight, about 30 % by weight, about 40 %
by
weight, about 50 % by weight, about 60 % by weight, about 70 % by weight,
about 80
% by weight, about 90 % by weight, about 95 % by weight, or about 100 A by
weight.
[161] In some embodiments, the composition includes a psychoactive alkaloid
extract having a total dephosphorylated psychoactive alkaloid content of a
trace by
weight to about 100 % by weight. In some embodiments, the composition includes
a
psychoactive alkaloid extract having a total dephosphorylated psychoactive
alkaloid
content of a trace by weight to about 10 % by weight, a trace by weight to
about 20 %
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by weight, a trace by weight to about 30 % by weight, a trace by weight to
about 40 %
by weight, a trace by weight to about 50 % by weight, a trace by weight to
about 60 %
by weight, a trace by weight to about 70 % by weight, a trace by weight to
about 80 %
by weight, a trace by weight to about 90 % by weight, a trace by weight to
about 95 %
by weight, a trace by weight to about 100 % by weight, about 10 % by weight to
about
20 % by weight, about 10 % by weight to about 30 % by weight, about 10 % by
weight
to about 40 % by weight, about 10 % by weight to about 50 % by weight, about
10 % by
weight to about 60 A by weight, about 10 % by weight to about 70 A by
weight, about
% by weight to about 80 % by weight, about 10 % by weight to about 90 A) by
weight, about 10 A by weight to about 95 % by weight, about 10 A by weight
to about
100 % by weight, about 20 % by weight to about 30 % by weight, about 20 % by
weight
to about 40 % by weight, about 20 % by weight to about 50 % by weight, about
20 % by
weight to about 60 A by weight, about 20 A by weight to about 70 A by
weight, about
% by weight to about 80 % by weight, about 20 % by weight to about 90 A) by
weight, about 20 % by weight to about 95 % by weight, about 20 A by weight to
about
100 % by weight, about 30 % by weight to about 40 % by weight, about 30 % by
weight
to about 50 % by weight, about 30 % by weight to about 60 % by weight, about
30 % by
weight to about 70 % by weight, about 30 % by weight to about 80 % by weight,
about
% by weight to about 90 % by weight, about 30 A by weight to about 95 % by
weight, about 30 % by weight to about 100 % by weight, about 40 % by weight to
about
50 A) by weight, about 40 % by weight to about 60 A by weight, about 40 % by
weight
to about 70 % by weight, about 40 % by weight to about 80 % by weight, about
40 % by
weight to about 90 % by weight, about 40 % by weight to about 95 A by weight,
about
% by weight to about 100 % by weight, about 50 % by weight to about 60 % by
weight, about 50 hi by weight to about 70 % by weight, about 50 A) by weight
to about
80 A) by weight, about 50 % by weight to about 90 A by weight, about 50 % by
weight
to about 95 % by weight, about 50 % by weight to about 100 % by weight, about
60 %
by weight to about 70 % by weight, about 60 % by weight to about 80 % by
weight,
about 60 % by weight to about 90 % by weight, about 60 % by weight to about 95
% by
weight, about 60 % by weight to about 100 % by weight, about 70 % by weight to
about
80 % by weight, about 70 % by weight to about 90 A by weight, about 70 % by
weight
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to about 95 % by weight, about 70 % by weight to about 100 % by weight, about
80 %
by weight to about 90 % by weight, about 80 % by weight to about 95 % by
weight,
about 80 A by weight to about 100 % by weight, about 90 % by weight to about
95 %
by weight, about 90 % by weight to about 100 % by weight, or about 95 % by
weight to
about 100 % by weight. In some embodiments, the composition includes a
psychoactive alkaloid extract having a total dephosphorylated psychoactive
alkaloid
content of a trace by weight, about 10 % by weight, about 20 % by weight,
about 30 %
by weight, about 40 A by weight, about 50 A) by weight, about 60 % by
weight, about
70 % by weight, about 80 % by weight, about 90 A) by weight, about 95 % by
weight, or
about 100 A by weight. In some embodiments, the composition includes a
psychoactive alkaloid extract having a total dephosphorylated psychoactive
alkaloid
content of at least a trace by weight, about 10 % by weight, about 20 % by
weight,
about 30 A by weight, about 40 % by weight, about 50 A by weight, about 60 %
by
weight, about 70 % by weight, about 80 % by weight, about 90 % by weight, or
about
95 A by weight. In some embodiments, the composition includes a psychoactive
alkaloid extract having a total dephosphorylated psychoactive alkaloid content
of at
most about 10 % by weight, about 20 % by weight, about 30 A by weight, about
40 %
by weight, about 50 % by weight, about 60 % by weight, about 70 % by weight,
about
80 % by weight, about 90 % by weight, about 95 % by weight, or about 100 % by
weight.
[162] In one embodiment, one psychoactive alkaloid extract and another
psychoactive alkaloid extract are present in a proportion such that the
specific ratio of
phosphorylated to dephosphorylated psychoactive alkaloid ranges from 1:1000 to
1000:1. In exemplary embodiments, a phosphorylated psychoactive alkaloid
extract
and a dephosphorylated psychoactive alkaloid extract are present in a
proportion such
that the specific ratio of phosphorylated to dephosphorylated psychoactive
alkaloid is
1:1, 1:3, and 3:1.
[163] In other embodiments, the psychoactive alkaloid composition is
defined by a
specific total amount of psychoactive alkaloid content, which has a known
ratio of
phosphorylated and dephosphorylated alkaloids.
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[164] In some embodiments, the psychoactive alkaloid composition with a
specific
ratio of a phosphorylated psychoactive alkaloid and a dephosphorylated
psychoactive
alkaloid is made by the process for obtaining a psychoactive alkaloid
composition with a
specific ratio of a phosphorylated psychoactive alkaloid to a dephosphorylated
psychoactive alkaloid of the present invention.
[165] The psychoactive alkaloid extract of the composition low in (e.g.,
<20%) or
almost free of undesired impurities allows a smaller amount of the composition
to
achieve a desired therapeutic effect than if the extract were less
concentrated, with
more impurities.
[166] The psychoactive alkaloid composition of the present invention is in
powder
form. This free-flowing powder form allows the composition to be easily
handled. The
components of the composition are also in powder form.
[167] The psychoactive alkaloid composition of the present invention can be
used,
for example, in medical research on the use of psychedelic substances in
treatments for
mental illnesses.
[168] FIG. 22, in another embodiment, describes a process for preparing a
psychoactive alkaloid composition having a phosphorylated psychoactive
alkaloid from
one psychoactive alkaloid extraction and a dephosphorylated psychoactive
alkaloid
from another psychoactive alkaloid extraction in a specific ratio.
[169] A dried powdered biomass is obtained in step 2201. Step 2202 includes
extracting a psychoactive alkaloid with a basified solvent to obtain a
psychoactive
alkaloid liquid with a pH greater than 10.5. A majority, or all, of a total
phosphorylatable
psychoactive alkaloid content is phosphorylated alkaloid and a remainder
thereof is
dephosphorylated alkaloid.
[170] In step 2203 the pH of the resulting psychoactive alkaloid liquid is
adjusted to
a pH ranging from 3.5 to 4.5. The pH is adjusted by adding an acid.
[171] Another dried powdered biomass is obtained in step 2206. Step 2207
includes extracting a psychoactive alkaloid with an acidified solvent to
obtain another
psychoactive alkaloid liquid with a pH lower than 3.5. All of the psychoactive
alkaloid
present is dephosphorylated.
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[172] Step 2208 involves adjusting the pH of the second psychoactive
alkaloid liquid
to a pH ranging from 3.5 to 4.5. The pH is adjusted by adding a base.
[173] In steps 2204 and 2209, a portion of the basified solvent from the
first
psychoactive alkaloid liquid and a portion of the acidified solvent from the
second
psychoactive alkaloid liquid are evaporated to obtain first and second
psychoactive
alkaloid slurries 2205 and 2210 respectively.
[174] In step 2211, measured portions of the psychoactive alkaloid slurries
are
mixed to obtain a bulk psychoactive alkaloid slurry. The bulk psychoactive
alkaloid
slurry includes the phosphorylated psychoactive alkaloid and the
dephosphorylated
psychoactive alkaloid in the specific ratio. The measured portions, by weight,
of each of
the psychoactive alkaloid slurries are chosen in such a manner as to achieve
the
specific ratio of the phosphorylated psychoactive alkaloid and the
dephosphorylated
psychoactive alkaloid, taking into account the percentage by weight of these
psychoactive alkaloids in each of the initial slurries. In one exemplary
embodiment,
mixing 100 g of a 1.0% phosphorylated psychoactive alkaloid slurry, with 50 g
of a 0.5%
dephosphorylated psychoactive alkaloid slurry results in the specific ratio of
4:1 of
phosphorylated to dephosphorylated alkaloid.
[175] In step 2212, the obtained bulk psychoactive alkaloid slurry is
standardized by
adding thereto a measured quantity of one or more excipients to obtain a
standardized
bulk slurry with a specific total amount of psychoactive alkaloid.
[176] Step 2213 includes drying the standardized bulk psychoactive alkaloid
slurry
by evaporation to obtain the psychoactive alkaloid composition. The obtained
psychoactive alkaloid composition has a specific total amount of psychoactive
alkaloid
content, and the phosphorylated psychoactive alkaloid and dephosphorylated
psychoactive alkaloid are in the specific ratio. In some embodiments of the
psychoactive alkaloid composition, the specific ratio of phosphorylated
psychoactive
alkaloid to dephosphorylated psychoactive alkaloid ranges from 1:1000 to
1000:1.
Mixture
[177] Referring to FIG. 21, in one embodiment, the present invention also
relates to
a process for preparing a psychoactive alkaloid composition by mixing a
phosphorylated psychoactive alkaloid composition and a dephosphorylated
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psychoactive alkaloid composition in a specific ratio. In step 2100, the
psychoactive
alkaloid composition obtained after step 2020 (FIG. 20) is taken. The
psychoactive
alkaloid composition in step 2020 has the desired amount of the
dephosphorylated
psychoactive alkaloid of 100% by weight of the total phosphorylatable
psychoactive
alkaloid content in the psychoactive alkaloid extract. The aforesaid is
achieved by
extracting the psychoactive alkaloid from the dried powdered biomass with an
acidified
solvent having a specific pH lower than 3.5.
[178] In step 2110, another psychoactive alkaloid composition is obtained
according
to the process described above. This other psychoactive alkaloid composition
has a
desired amount of phosphorylated psychoactive alkaloid of 100% by weight of
the total
phosphorylatable psychoactive alkaloid content. The aforesaid is achieved by
extracting the psychoactive alkaloid from the dried powdered biomass with a
basified
solvent having a pH greater than 10.5, where the biomass is not
dephosphorylated or
not significantly dephosphorylated.
[179] In some embodiments, the acidified solvent is a mixture of an acid
and a C1-
C4 primary aliphatic alcohol, a C3-C4 ketone, water, or any combination
selected
therefrom. The acid may be citric acid, ascorbic acid, formic acid, acetic
acid,
hydrochloric acid, phosphoric acid, sulfuric acid, or any combination selected
therefrom.
In other embodiments, the basified solvent is a mixture of a base and a C1-C4
primary
aliphatic alcohol, a C3-C4 ketone, water, or any combination selected
therefrom. The
base may be sodium hydroxide, potassium hydroxide, ammonium hydroxide, sodium
bicarbonate, calcium carbonate, or any combination selected therefrom.
[180] In step 2120, both psychoactive alkaloid compositions are mixed in a
measured ratio to obtain a psychoactive alkaloid composition. This composition
includes the dephosphorylated psychoactive alkaloid of the first psychoactive
alkaloid
composition and the phosphorylated psychoactive alkaloid of the second
psychoactive
alkaloid composition in a specific ratio. In some embodiments, the specific
ratio of
phosphorylated psychoactive alkaloid to dephosphorylated psychoactive alkaloid
is in
the range from 1000:1 to 1:1000. If the total phosphorylatable psychoactive
alkaloid
concentrations are the same in both starting compositions, then the final
ratio of
phosphorylated to dephosphorylated alkaloids is the same as the ratio in which
the
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starting compositions are mixed. However, in other embodiments, the mixing
ratio may
need to be modified to take into account the different starting concentrations
of the
phosphorylated and dephosphorylated alkaloids in their respective
compositions. In yet
other embodiments, one of the starting compositions may include both
phosphorylated
and dephosphorylated alkaloids, and the mixing ratio of the two compositions
may be
adjusted to take this into account.
Naturally Occurring Substances
[181] In other embodiments, the psychoactive alkaloid extract includes
naturally
occurring substances selected from fats, sugars, carbohydrates, and proteins,
or any
combination selected therefrom. The aforesaid naturally occurring substances
do not
lead to any side effects or adverse effects when ingested as a part of the
composition.
[182] The naturally-occurring substances are present in the psychoactive
alkaloid
extract in a concentration ranging from 1-99.9% by dry weight. The
concentration
range of the naturally-occurring substances in the psychoactive alkaloid
extract will vary
due to various factors for example, but not limited to, the source of the
psychoactive
alkaloid extract, the extraction technique used, the efficiency of the
extraction process,
and the amount of the psychoactive alkaloid in the extract.
[183] In some embodiments, the psychoactive alkaloid extract includes the
psychoactive alkaloid and the naturally occurring substances.
Excipient
[184] The excipients described herein refer to excipients which aid in the
manufacturing and/or administration of the compositions described herein. Non-
limiting
examples of such excipients are well known in the art and include flavorants,
colorants,
palatants, antioxidants, viscosity modifying agents, tonicity agents, drug
carriers,
sustained-release agents, comfort-enhancing agents, emulsifiers, solubilizing
aids,
lubricants, binding agents, stabilizing agents and other agents to aid in the
manufacturing and/or administration of the compositions. The excipients used
in the
present invention are acceptable for use in pharmaceutical or nutraceutical
applications
or as food ingredients. The amount of excipients will vary depending on the
concentration of psychoactive alkaloids in the psychoactive alkaloid extract
to result in a
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flowable psychoactive alkaloid composition, which will be obvious to a person
of skill in
the art.
[185] In some embodiments, a bioavailability enhancing agent such as citric
acid,
beta cyclodextrin, or alpha cyclodextrin can be added (up to 0.5% by weight)
as an
excipient. In other embodiments, an antioxidant agent such as ascorbic acid
may be
added (up to 0.5% by weight) as an excipient.
[186] In some embodiments, the excipients are selected from silicon
dioxide,
ascorbic acid, maltodextrin from corn, potato, or tapioca for example, gum
arabic,
microcrystalline cellulose, sodium benzoate, sodium phosphate, sodium citrate,
rice
hulls, and rice. A combination of any of these excipients may be used.
[187] In some embodiments, the carrier comprises maltodextrin in the
composition
at 10% to 20%. In some embodiments, the carrier comprises maltodextrin in the
composition at about 10 % to about 12 `)/0, about 10 `)/0 to about 14 `)/0,
about 10 % to
about 16%, about 10% to about 18%, about 10% to about 20%, about 12% to about
14 `)/0, about 12 `)/0 to about 16 %, about 12 `)/0 to about 18 %, about 12 %
to about 20 %,
about 14 % to about 16 %, about 14 % to about 18 %, about 14 % to about 20 %,
about
16 % to about 18 %, about 16 % to about 20 %, or about 18 % to about 20 %. In
some
embodiments, the carrier comprises maltodextrin in the composition at about 10
%,
about 12%, about 14%, about 16%, about 18%, or about 20%. In some
embodiments, the carrier comprises maltodextrin in the composition at least
about 10
%, about 12%, about 14%, about 16%, or about 18%. In some embodiments, the
carrier comprises maltodextrin in the composition at most about 12 %, about 14
%,
about 16 %, about 18 %, or about 20 %.
[188] In some embodiments, the carrier comprises mannitol in the
composition at
10% to 20%. In some embodiments, the carrier comprises mannitol in the
composition
at about 10 % to about 12%, about 10% to about 14%, about 10% to about 16%,
about 10 /.0 to about 18 %, about 10 % to about 20 `)/0, about 12 `)/0 to
about 14 %, about
12 % to about 16 %, about 12 % to about 18 %, about 12 % to about 20 %, about
14 %
to about 16%, about 14% to about 18%, about 14% to about 20%, about 16% to
about 18 `)/0, about 16 % to about 20 %, or about 18 % to about 20 %. In some
embodiments, the carrier comprises mannitol in the composition at about 10 %,
about
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12 A), about 14 A, about 16 %, about 18 %, or about 20 %. In some
embodiments, the
carrier comprises mannitol in the composition at least about 10 %, about 12 %,
about
14 A, about 16 A, or about 18 %. In some embodiments, the carrier comprises
mannitol in the composition at most about 12 %, about 14 %, about 16 %, about
18 %,
or about 20 %.
[189] Other embodiments are also possible. While only specific neutralizing
agents,
food grade acids and food grade bases have been mentioned herein, other
neutralizing
agents, food grade acids and food grade bases may be used.
[190] In some embodiments, the excipient is a carrier, a flowability agent,
a
preservative or any combination selected therefrom. The amount of excipient in
the
composition will vary depending on the desired amount of the psychoactive
alkaloid
extract in the composition, and on the concentration of psychoactive alkaloids
in the
extract. It will also depend on the degree of flowability required and the
stability
required.
[191] In some embodiments, the excipients include a flowability agent, in
an amount
up to 2% by weight of the composition. Above 2%, little extra benefit is
gained. In other
embodiments, the excipients include a flowability agent in a concentration
equal to or
less than 2% by weight of the composition. In some embodiments, the
flowability agent
is present in the composition at, by weight, about 0.1 % to about 2 %. In some
embodiments, the flowability agent is present in the composition at, by
weight, about 2
% to about 1.75 %, about 2 % to about 1.5 %, about 2 A) to about 1.25 %,
about 2 % to
about 1 %, about 2 % to about 0.75 %, about 2 % to about 0.5 %, about 2 % to
about
0.25 %, about 2 A to about 0.1 %, about 1.75 A to about 1.5 %, about 1.75 A
to about
1.25%, about 1.75 % to about 1 %, about 1.75% to about 0.75 %, about 1.75 % to
about 0.5 %, about 1.75 % to about 0.25 A, about 1.75 hi to about 0.1 A,
about 1.5 %
to about 1.25 %, about 1.5 % to about 1 %, about 1.5 % to about 0.75 %, about
1.5 %
to about 0.5 A, about 1.5 A to about 0.25 %, about 1.5 % to about 0.1 %,
about 1.25 %
to about 1 %, about 1.25 % to about 0.75 %, about 1.25 % to about 0.5 %, about
1.25
% to about 0.25 A, about 1.25 % to about 0.1 A, about 1 % to about 0.75 %,
about 1 %
to about 0.5 A, about 1 % to about 0.25 %, about 1 % to about 0.1 %, about
0.75 % to
about 0.5 %, about 0.75 % to about 0.25 %, about 0.75 % to about 0.1 %, about
0.5 %
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to about 0.25 %, about 0.5 A to about 0.1 %, or about 0.25 % to about 0.1 %.
In some
embodiments, the flowability agent is present in the composition at, by
weight, about 2
%, about 1.75 A, about 1.5 %, about 1.25 %, about 1 %, about 0.75 %, about
0.5 %,
about 0.25 %, or about 0.1 %. In some embodiments, the flowability agent is
present in
the composition at, by weight, at least about 2 %, about 1.75 %, about 1.5 %,
about
1.25 A, about 1 %, about 0.75 %, about 0.5 A, or about 0.25 %. In some
embodiments, the flowability agent is present in the composition at, by
weight, at most
about 1.75 %, about 1.5 %, about 1.25 A, about 1 %, about 0.75 %, about 0.5
%, about
0.25 %, or about 0.1 %.
[192] In some embodiments, the flowability agent is silicon dioxide. In
other
embodiments, the flowability agent is magnesium stearate, stearic acid or
talc, or is
selected from any other known, suitable flowability agent. A combination of
any of
these flowability agents may be used. It is also envisaged that other
flowability agents
can be used. The impurities present in the extract tend to have a negative
effect on the
flowability, and flow agents are added to counter these effects. While there
are known
methods of measuring flowability, this is not always necessary as the
product's lack of
adequate flowability is often very evident when one handles the product.
[193] In some embodiments, the excipients include a carrier, in an amount
up to
94% by weight of the composition. In some embodiments, the excipients include
a
carrier ranging from 10-94% by weight of the composition. In some embodiments,
the
carrier is present in the composition at, by weight, about 10 % to about 94 %.
In some
embodiments, the carrier is present in the composition at, by weight, about 10
% to
about 20 A, about 10 % to about 30 %, about 10 A to about 40 %, about 10 %
to about
50%, about 10% to about 60%, about 10% to about 70%, about 10% to about 80%,
about 10 A to about 90 A, about 10 hi to about 94 %, about 20 % to about 30
%, about
20 % to about 40 %, about 20 % to about 50 %, about 20 A) to about 60 %,
about 20 %
to about 70 A, about 20 % to about 80 %, about 20 % to about 90 %, about 20 %
to
about 94 %, about 30 % to about 40 %, about 30 % to about 50 %, about 30 % to
about
60 %, about 30 % to about 70 %, about 30 % to about 80 %, about 30 % to about
90 %,
about 30 A to about 94 %, about 40 % to about 50 %, about 40 A to about 60
%, about
40 % to about 70 %, about 40 % to about 80 %, about 40 % to about 90 %, about
40 %
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to about 94 %, about 50 % to about 60 %, about 50 % to about 70 %, about 50 %
to
about 80 %, about 50 % to about 90 %, about 50 % to about 94 %, about 60 A to
about
70 %, about 60 % to about 80 %, about 60 % to about 90 %, about 60 A to about
94 A,
about 70 % to about 80 %, about 70 % to about 90 %, about 70 % to about 94 %,
about
80 % to about 90 %, about 80 % to about 94 %, or about 90 % to about 94 %. In
some
embodiments, the carrier is present in the composition at, by weight, about 10
%, about
20 %, about 30 %, about 40 %, about 50 %, about 60 %, about 70 %, about 80 %,
about 90 %, or about 94 %. In some embodiments, the carrier is present in the
composition at, by weight, at least about 10 %, about 20 %, about 30 %, about
40 %,
about 50 %, about 60 %, about 70 %, about 80 %, or about 90 %. In some
embodiments, the carrier is present in the composition at, by weight, at most
about 20
%, about 30 %, about 40 %, about 50 %, about 60 %, about 70 %, about 80 %,
about
90 `)/0, or about 94 `)/0.
[194] In some embodiments, the carrier is mannitol, maltodextrin, or in
other
embodiments it is microcrystalline cellulose, coconut flour or corn starch, or
any other
known, suitable carrier. A combination of any of these carriers may be used.
It is also
envisaged that other carriers can be used.
[195] The preservative is selected from ascorbic acid, citric acid,
lactose, vitamin A,
vitamin E, retinyl palmitate, selenium, sodium citrate, sodium ascorbate,
calcium
ascorbate, sodium benzoate, and potassium benzoate. A combination of any of
these
preservatives may be used. It is also envisaged that other preservatives can
be used.
[196] In some embodiments, the excipients include a preservative, up to 10%
by
weight of the composition. In some embodiments, preservatives are not added to
the
composition. There may be cases where a preservative is not required, or not
wanted
in the final product, and the concentration of preservative will be 0%. At a
certain point
(>10% in this example), the preservative will not give any further benefit,
and so the
upper limit of preservative is 10%.In some embodiments, the preservatives are
present
in the composition at, by weight, about 0.1 "Yo to about 10 %. In some
embodiments,
the preservatives are present in the composition at, by weight, about 10 % to
about 9
%, about 10% to about 8%, about 10% to about 7%, about 10% to about 6%, about
% to about 5 %, about 10 % to about 4 %, about 10 % to about 3 %, about 10 %
to
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about 2%, about 10 % to about 1 %, about 10 % to about 0.5%, about 10 % to
about
0.1 %, about 9 % to about 8 %, about 9 % to about 7 %, about 9 % to about 6 %,
about
9 % to about 5 %, about 9 % to about 4 %, about 9 % to about 3 %, about 9 A
to about
2 %, about 9 "Yo to about 1 %, about 9 % to about 0.5 %, about 9 "Yo to about
0.1 %,
about 8 % to about 7 %, about 8 % to about 6 %, about 8 % to about 5 %, about
8 % to
about 4 %, about 8 % to about 3 %, about 8 % to about 2 %, about 8 % to about
1 %,
about 8 % to about 0.5 %, about 8 % to about 0.1 %, about 7 % to about 6 %,
about 7
% to about 5 %, about 7 % to about 4 `)/0, about 7 % to about 3 %, about 7 %
to about 2
%, about 7 % to about 1 %, about 7 % to about 0.5 %, about 7 % to about 0.1 %,
about
6 % to about 5 %, about 6 % to about 4 %, about 6 % to about 3 %, about 6 % to
about
2 %, about 6 % to about 1 %, about 6 % to about 0.5 %, about 6 % to about 0.1
%,
about 5 % to about 4 /0, about 5 % to about 3 %, about 5 % to about 2 %,
about 5 % to
about 1 %, about 5 % to about 0.5 %, about 5 `)/0 to about 0.1 %, about 4 % to
about 3
%, about 4 % to about 2 %, about 4 % to about 1 %, about 4 % to about 0.5 %,
about 4
% to about 0.1 %, about 3 % to about 2 %, about 3 A to about 1 %, about 3 %
to about
0.5 %, about 3 % to about 0.1 %, about 2 % to about 1 %, about 2 % to about
0.5 %,
about 2 % to about 0.1 %, about 1 % to about 0.5 %, about 1 % to about 0.1 %,
or
about 0.5 % to about 0.1 %. In some embodiments, the preservatives are present
in
the composition at, by weight, about 10 %, about 9 %, about 8 %, about 7 %,
about 6
%, about 5 %, about 4 %, about 3 `)/0, about 2 %, about 1 %, about 0.5 %, or
about 0.1
%. In some embodiments, the preservatives are present in the composition at,
by
weight, at least about 10 %, about 9 %, about 8 %, about 7 `)/0, about 6 %,
about 5 %,
about 4 %, about 3 %, about 2 %, about 1 %, or about 0.5 %. In some
embodiments,
the preservatives are present in the composition at, by weight, at most about
9 A, about
8 %, about 7 %, about 6 %, about 5 `)/0, about 4 %, about 3 %, about 2 %,
about 1 `)/0,
about 0.5 %, or about 0.1 %.
[197] In some embodiments, a psychoactive alkaloid composition comprises
one or
more preservatives up to 10%.
[198] In some embodiments, a first preservative of the one or more
preservatives is
present in the composition from about 0.1 % to about 3 %. In some embodiments,
a first
preservative of the one or more preservatives is present in the composition
from about
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0.1 % to about 0.5%, about 0.1 A to about 0.9%, about 0.1 A to about 1.3%,
about
0.1 % to about 1.7 %, about 0.1 A to about 2.1 A, about 0.1 % to about 2.4
%, about
0.1 % to about 2.7 A, about 0.1 % to about 3 %, about 0.5 % to about 0.9 A,
about 0.5
% to about 1.3 %, about 0.5 A to about 1.7 A, about 0.5 % to about 2.1 %,
about 0.5 %
to about 2.4 %, about 0.5 % to about 2.7 A, about 0.5 % to about 3 %, about
0.9 % to
about 1.3 %, about 0.9 % to about 1.7 %, about 0.9 A to about 2.1 %, about
0.9 A to
about 2.4 %, about 0.9 % to about 2.7 %, about 0.9 A to about 3 %, about 1.3
% to
about 1.7 %, about 1.3 % to about 2.1 %, about 1.3 % to about 2.4 %, about 1.3
% to
about 2.7 %, about 1.3 % to about 3 %, about 1.7 A to about 2.1 A, about 1.7
% to
about 2.4 %, about 1.7 % to about 2.7 %, about 1.7 % to about 3 %, about 2.1 %
to
about 2.4 %, about 2.1 % to about 2.7 %, about 2.1 % to about 3 %, about 2.4 %
to
about 2.7 A, about 2.4 % to about 3 %, or about 2.7 A to about 3 %. In some
embodiments, a first preservative of the one or more preservatives is present
in the
composition from about 0.1 A, about 0.5 %, about 0.9 %, about 1.3 %, about
1.7 A,
about 2.1 A, about 2.4 A, about 2.7 A, or about 3 %. In some embodiments, a
first
preservative of the one or more preservatives is present in the composition
from at least
about 0.1 %, about 0.5 %, about 0.9 A, about 1.3 A, about 1.7 %, about 2.1
A, about
2.4 %, or about 2.7 %. In some embodiments, a first preservative of the one or
more
preservatives is present in the composition from at most about 0.5 A, about
0.9 %,
about 1.3 A, about 1.7%, about 2.1 %, about 2.4 A, about 2.7 %, or about 3
%.
[199] In
some embodiments, a first preservative of the one or more preservatives is
present in the composition from about 0.1 % to about 2 %. In some embodiments,
a first
preservative of the one or more preservatives is present in the composition
from about
0.1 % to about 0.3 %, about 0.1 A to about 0.5 A, about 0.1 % to about 0.7
%, about
0.1 % to about 0.9 A, about 0.1 % to about 1.1 %, about 0.1 A to about 1.3
%, about
0.1 % to about 1.5 %, about 0.1 % to about 1.7 A, about 0.1 % to about 2 A,
about 0.3
% to about 0.5 %, about 0.3 % to about 0.7 A, about 0.3 A to about 0.9 %,
about 0.3 %
to about 1.1 %, about 0.3 % to about 1.3 A, about 0.3 % to about 1.5 A,
about 0.3 % to
about 1.7 %, about 0.3 % to about 2 A, about 0.5 A to about 0.7 %, about 0.5
% to
about 0.9 %, about 0.5 % to about 1.1 %, about 0.5 % to about 1.3 %, about 0.5
% to
about 1.5 %, about 0.5 % to about 1.7 %, about 0.5 A to about 2 %, about 0.7
% to
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about 0.9 %, about 0.7 % to about 1.1 %, about 0.7 % to about 1.3 %, about 0.7
% to
about 1.5 %, about 0.7 % to about 1.7 %, about 0.7 % to about 2 %, about 0.9 %
to
about 1.1 %, about 0.9 % to about 1.3 %, about 0.9 % to about 1.5 A, about
0.9 % to
about 1.7 %, about 0.9 % to about 2 %, about 1.1 % to about 1.3 %, about 1.1 %
to
about 1.5%, about 1.1 % to about 1.7%, about 1.1 % to about 2%, about 1.3% to
about 1.5 %, about 1.3 % to about 1.7 %, about 1.3 % to about 2 %, about 1.5 %
to
about 1.7 %, about 1.5 % to about 2 %, or about 1.7 % to about 2 %. In some
embodiments, a first preservative of the one or more preservatives is present
in the
composition from about 0.1 %, about 0.3 %, about 0.5 %, about 0.7 %, about 0.9
%,
about 1.1 %, about 1.3 /.0, about 1.5 %, about 1.7 %, or about 2 %. In some
embodiments, a first preservative of the one or more preservatives is present
in the
composition from at least about 0.1 %, about 0.3 %, about 0.5 %, about 0.7 %,
about
0.9 %, about 1.1 %, about 1.3 %, about 1.5 %, or about 1.7 `)/0. In some
embodiments, a
first preservative of the one or more preservatives is present in the
composition from at
most about 0.3 A, about 0.5 `)/0, about 0.7 `)/0, about 0.9 %, about 1.1 %,
about 1.3 %,
about 1.5%, about 1.7%, or about 2 %.
[200] In some embodiments, a second preservative of the one or more
preservatives is present in the composition from about 0.1 % to about 2 %. In
some
embodiments, a second preservative of the one or more preservatives is present
in the
composition from about 0.1 % to about 0.3 %, about 0.1 % to about 0.5 %, about
0.1 %
to about 0.7 %, about 0.1 % to about 0.9 %, about 0.1 % to about 1.1 %, about
0.1 % to
about 1.3 %, about 0.1 % to about 1.5 %, about 0.1 % to about 1.7 c1/0, about
0.1 % to
about 2 %, about 0.3 % to about 0.5 %, about 0.3 % to about 0.7 %, about 0.3 %
to
about 0.9 %, about 0.3 % to about 1.1 %, about 0.3 % to about 1.3 %, about 0.3
% to
about 1.5 %, about 0.3 % to about 1.7 %, about 0.3 % to about 2 %, about 0.5 %
to
about 0.7%, about 0.5% to about 0.9%, about 0.5% to about 1.1 %, about 0.5% to
about 1.3 %, about 0.5 % to about 1.5 %, about 0.5 % to about 1.7 %, about 0.5
% to
about 2%, about 0.7% to about 0.9%, about 0.7% to about 1.1 %, about 0.7% to
about 1.3 %, about 0.7 % to about 1.5 %, about 0.7 % to about 1.7 %, about 0.7
% to
about 2 %, about 0.9 % to about 1.1 %, about 0.9 `)/0 to about 1.3 %, about
0.9 % to
about 1.5 %, about 0.9 % to about 1.7 %, about 0.9 % to about 2 %, about 1.1 %
to
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about 1.3%, about 1.1 % to about 1.5%, about 1.1 % to about 1.7%, about 1.1 %
to
about 2 %, about 1.3 A) to about 1.5 %, about 1.3 % to about 1.7 %, about 1.3
% to
about 2 %, about 1.5 % to about 1.7 %, about 1.5 % to about 2 %, or about 1.7
% to
about 2 %. In some embodiments, a second preservative of the one or more
preservatives is present in the composition from about 0.1 %, about 0.3 %,
about 0.5 %,
about 0.7%, about 0.9%, about 1.1 %, about 1.3%, about 1.5%, about 1.7%, or
about 2 %. In some embodiments, a second preservative of the one or more
preservatives is present in the composition from at least about 0.1 %, about
0.3 A,
about 0.5 %, about 0.7 %, about 0.9 %, about 1.1 %, about 1.3 %, about 1.5 %,
or
about 1.7 %. In some embodiments, a second preservative of the one or more
preservatives is present in the composition from at most about 0.3 %, about
0.5 %,
about 0.7%, about 0.9%, about 1.1 %, about 1.3%, about 1.5%, about 1.7%, or
about 2 %.
[201] The main consideration is that, depending on the source material's
concentration and the efficiency of the extraction, the concentration of the
extract needs
to be blended down to the required value of standardization. This dictates the
amount
of excipient for each batch, which may be different. Therefore, the amount of
excipient
that can be added can be anywhere between say 0.100% to 99.9%, depending on
the
source concentration before blending. The other excipient components may be
held
relatively constant between batches, or within a much narrower range (i.e., 0-
2%
flowability agent, 0-10% preservative, 0-0.5% antioxidant, or 0-0.5%
bioavailability
agent).
C. Apparatus
[202] Referring to FIG. 8, an example of the apparatus is shown
schematically.
Raw psilocybin mushrooms are provided in a hopper 800, for example, and are
released in batches into container 801. The raw fungal material is then dried
in a forced
air oven 802. The dried biomass is placed into a grinder 803 for grinding.
[203] After the drying and grinding steps, the ground biomass is placed in
an
agitated, heat-controlled extraction vessel 805. The vessel holds the biomass
and
solvent 806, such as lower aliphatic alcohols, water, buffered acid or
buffered alkaline,
or a mixture thereof. The vessel may be surrounded by an insulating wall 804.
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Alternately, there may be an insulating jacket wrapped around the vessel. The
insulating wall 804 or jacket helps to maintain the contents 806 under a
constant
temperature (T) between 5-95 C. The pressure (P) inside the extraction vessel
805
may be regulated up to 100 MPa (15000 psig).
[204] After the extraction, the bottom of the extraction vessel 805 is
opened at outlet
807, and the extraction slurry is collected in container 808. The extraction
slurry is then
fed into filter 809. After filtration, the first filtrate leaves the filter
809 and is collected in
container 810. The first filtrate residue 811 is then fed back at R into
agitated, heat-
controlled vessel 805 and more solvent (S) is added. After the second
extraction, the
extraction slurry is collected in container 808 and is then fed into filter
812 (or filter 809).
After filtration, the second filtrate and solvent mixture leaves the filter
812 and is
collected in container 813.
[205] After the two filtration stages, if there are two, the filtrates are
mixed in
container 814. Otherwise, if there is only a single filtration step, mixing is
unnecessary.
Neutralizer is added as necessary to the filtrate in container 814. The
extraction slurry,
pH-adjusted where necessary, is then passed to rotary evaporator 815 in which
all or
part of the solvent is evaporated, depending on the embodiment. If all the
solvent is
evaporated, then reverse osmosis water is added to the solids remaining after
the
evaporation.
[206] The concentrated slurry is then passed to container 816 and tested to
determine its alkaloid content, using a titration setup 817. Carriers are
added to
container 816 with the concentrated slurry, and mixed. The standardized slurry
is then
placed in a bench-top spray drier 818 to produce psilocybin mushroom extract
that is
collected in container 819.
[207] Referring to FIG. 13, an example of the apparatus is shown
schematically.
Raw Psilocybe cubensis mushrooms are added to a hopper 1300 and then released
in
batches into container 1301. The raw fungal material is then dried in a forced
air oven
1302 to result in dried biomass. The dried biomass is placed into a grinder
1303 for
grinding.
[208] The dried powdered biomass is placed into a heat-controlled vessel
1305, and
solvent (S) is added to the heat-controlled vessel. The vessel 1305 is
surrounded by an
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insulating wall 1304. Alternately, an insulating jacket may be wrapped around
the
vessel. The insulating wall 1304 or jacket helps to maintain the contents 1306
under a
constant temperature (T) between 5-95 C. The pressure (P) inside the
extraction
vessel 1305 may be regulated up to 100 MPa (15,000 psig).
[209] After the extraction, the bottom of the extraction vessel 1305 was
opened at
outlet 1307 and the extraction slurry was collected in a container 1308. The
extraction
slurry was then fed into a filter 1309 and a first filtrate was collected in
container 1310.
The first filtrate residue 1311 was then fed back (R) into the agitated, heat-
controlled
vessel 1305 and more solvent (S) was added for a second extraction. After the
second
extraction, the extraction slurry was collected in the container 1308 and was
then fed
into a filter 1312. After filtration, the obtained second filtrate was
collected in container
1313.
[210] After the two filtration stages, the filtrates were mixed in
container 1314 to
obtain a bulk filtrate. In other embodiments, if there is only a single
filtration step, this
mixing step is not required.
[211] The bulk filtrate is placed in a rotary evaporator 1315, and part of
the solvent
is evaporated from the bulk filtrate. The resultant extract is transferred to
a container
1316, where the pH of the extract is adjusted, followed by centrifugation 1317
to
remove the solid precipitates. The resultant supernatant is loaded onto a
column 1318
of resin. An initial wash is given to the column with a solvent to remove
impurities from
the resin, and fraction 1319 is collected. A second wash is given to the
column with
another solvent to elute the psychoactive alkaloids from the column and
results in
fraction 1320. A final wash is given to the column with another solvent to
wash any
impurities from the column and to prepare the column for use again, and the
fraction
1321 is obtained. The elution fraction 1320 with the psychoactive alkaloids is
then
concentrated in a rotary evaporator 1322 to result in the purified
psychoactive alkaloid
solution.
[212] In a container 1323 the purified psychoactive alkaloid solution and
desired
excipients are added together and thoroughly mixed to result in a final
standardized
slurry having a specified concentration of alkaloids. The final standardized
slurry is
then subjected to spray-drying 1324 to obtain a final powdered alkaloid
extract 1325
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with a total psilocybin/psilocin concentration defined as a percentage to two
decimal
places or two significant figures by dry weight.
[213] In other embodiments, parts of the apparatus may be reused or
duplicated.
For example, if desired, the elution fraction 1320 may be reloaded into the
container
1316 for pH adjustment, and the steps from thereon can be repeated to allow
for further
purification of the obtained purified psychoactive alkaloid solution.
[214] Referring to FIG. 17, an example of the apparatus is shown
schematically.
Raw Psilocybe cubensis mushrooms are added to a hopper 1700 and released in
batches into container 1701. The raw fungal material is then dried in a forced
air oven
1702 to result in dried biomass. The dried biomass is placed into a grinder
1703 for
grinding.
[215] The dried powdered biomass is placed into a heat-controlled vessel
1705, and
solvent (S) is added to the heat-controlled vessel. The vessel 1705 is
surrounded by an
insulating wall 1704. Alternately, an insulating jacket can be wrapped around
the
vessel. The insulating wall 1704 or jacket helps to maintain the contents 1706
under a
constant temperature (T) between 5-95 C. The pressure (P) inside the
extraction
vessel 1705 may be regulated up to 100 MPa (15,000 psig).
[216] After the extraction, the bottom of the extraction vessel 1705 is
opened at
outlet 1707 and the extraction slurry is collected in a container 1708. The
extraction
slurry is then fed into a filter 1709, and a first filtrate is collected in
container 1710. The
first filtrate residue 1711 is then fed back (R) into the agitated, heat-
controlled vessel
1705, and more solvent (S) is added for a second extraction. After the second
extraction, the extraction slurry is collected in the container 1708 and is
then fed into a
filter 1712. After filtration, the obtained second filtrate is collected in
container 1713.
[217] After the two filtration stages, the filtrates are mixed in container
1714 to
obtain a bulk filtrate. In other embodiments, if there is only a single
filtration step, this
mixing step is not required.
[218] The bulk filtrate was placed in a rotary evaporator 1715, and part of
the
solvent is evaporated from the bulk filtrate. The resultant extract is
transferred to a
container 1716, where the pH of the extract is adjusted, followed by
centrifugation 1717
to remove the solid precipitates. The resultant supernatant is loaded onto a
column
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Date Recue/Date Received 2022-07-25
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1718 of resin. An initial wash is given to the column with a solvent to remove
impurities
from the resin, and fraction 1719 is collected. A second wash is given to the
column
with another solvent to elute the psychoactive alkaloids from the column and
result in
fraction 1720. A final wash is given to the column with another solvent to
wash any
impurities from the column and to prepare the column for use again, and the
fraction
1721 was obtained. The elution fraction 1720 with the psychoactive alkaloids
is then
concentrated in a rotary evaporator 1722 to result in the purified
psychoactive alkaloid
extract.
[219] In a container 1723 the purified psychoactive alkaloid extract and
desired
excipients are added together and thoroughly mixed to result in a final
standardized
slurry having a specific amount of alkaloids. The final standardized slurry is
then
subjected to spray-drying 1724 to obtain a final powdered composition 1725
with a total
psilocybin/psilocin concentration defined as a percentage to two decimal
places or two
significant figures by dry weight.
[220] In other embodiments, parts of the apparatus may be reused or
duplicated.
For example, if desired, the elution fraction 1720 may be reloaded into the
container
1716 for pH adjustment, and the steps from thereon can be repeated to allow
for further
purification of the obtained purified psychoactive alkaloid extract.
[221] In one embodiment, the psychoactive alkaloid extract is obtained
after
evaporating the solvent from the bulk filtrate. The psychoactive alkaloid
extract is
transferred to a container 1723, and desired excipients are added together,
and
thoroughly mixed to result in a final standardized slurry having a specific
amount of
alkaloids. The final standardized slurry is then subjected to spray-drying
1724 to obtain
a final powdered composition 1725 with a total psilocybin/psilocin
concentration defined
as a percentage to two decimal places or two significant figures by dry
weight.
[222] Referring to FIG. 23, an example of the apparatus is shown
schematically.
Raw Psilocybe cubensis mushrooms are added to a hopper 2300 and then released
in
batches into container 2301. The raw fungal material is then dried in a forced
air oven
2302 to result in the dried biomass. The dried biomass is placed into a
grinder 2303 for
grinding to result in dried powdered biomass.
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[223] The dried powdered biomass is placed into a heat-controlled vessel
2305, and
acidified/basified solvent (S) is added to the vessel to obtain a specific pH
(lower than
3.5 or greater than 10.5). The vessel 2305 is surrounded by an insulating wall
2304.
Alternately, an insulating jacket may be wrapped around the vessel. The
insulating wall
108 or jacket helps to maintain the contents 2306 under a constant temperature
(T)
between 5-95 C. The pressure (P) inside the extraction vessel 2305 may be
regulated
from 7 to 20,000 psi. The extraction is performed with a solvent to solid
(dried
powdered biomass) proportion in the range of 1L:1kg to 50L:1kg.
[224] After the extraction, the bottom of the extraction vessel 2305 is
opened at
outlet 2307, and the extraction slurry is collected in a container 2308. The
extraction
slurry is then fed into a filter 2309, and a first filtrate is collected in
container 2310. The
first filtrate residue 2311 is then fed back (R) into the agitated, heat-
controlled vessel
2305, and more solvent (S) is added for a second extraction. After the second
extraction, the extraction slurry is collected in the container 2308 and then
fed into a
filter 2312. After filtration, the obtained second filtrate is collected in
container 2313.
[225] After the two filtration stages, the filtrates are mixed in container
2314 to
obtain a mixed filtrate, i.e., the psychoactive alkaloid liquid. In other
embodiments, if
there is only a single filtration step, this mixing step is not required. By
adding an acid
or a base, the pH of the psychoactive alkaloid liquid is brought to a pH
ranging from
3.5-4.5.
[226] The pH-adjusted, mixed filtrate is then placed in a rotary evaporator
2315, and
part of the solvent is evaporated from the mixed filtrate to form the
psychoactive
alkaloid extract, which is here a slurry.
[227] For obtaining the psychoactive alkaloid composition, the evaporation
in the
rotary evaporator 2315 is stopped after a desired portion of solvent is
evaporated. The
resultant slurry is transferred to a container 2316 where a measured quantity
of one or
more excipients is added to obtain a standardized slurry. The obtained
standardized
slurry is dried in a freeze dryer 2317 to obtain the psychoactive alkaloid
composition.
[228] In other embodiments, parts of the apparatus may be reused or
duplicated.
For example, if desired, for further extraction, the filtrate residue may be
reloaded into
the extraction vessel 2305, and the obtained filtrate can be added to mixed
filtrate
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container of 2314, and the steps from thereon can be repeated to obtain the
psychoactive alkaloid extract.
D. Extraction
Extraction Process 1
[229] Referring to FIG. 1, a flowchart is shown of the basic steps of the
extraction
process for extracting psychoactive compounds from psychedelic fungus. In step
100,
a solvent is added to a biomass of dried and ground, raw psychedelic fungus.
The raw
psychedelic fungus includes psilocybin fungi, including Psilocybe cubensis
mushrooms,
Psilocybe cyanescens mushrooms, Amanita muscaria mushrooms or a mixture of
these. Other species of psychedelic mushrooms may also be used.
[230] The parts of the mushrooms used include, for example, caps, gills,
stems, and
hyphae, and more particularly, any part of the psilocybin mushroom or mycelium
can be
included. In other cases, the raw psychedelic fungus parts used include only
caps, or
only stems, or only gills, or only hyphae or only mycelium or any mixture
thereof. In still
other cases, parts of the raw psychedelic fungus used are those that would
normally be
considered waste, in which valuable psychoactive compounds are found only in
lower
concentrations. The mushroom parts may be ground using a milling machine or
pulverization device, for example.
[231] Ideally, the moisture content of the raw plant material after drying
is low
compared to the total dried biomass weight. For example, the moisture content
may be
under 5% for smaller scale extractions and under 10% for larger scale
extractions. Wet
mushrooms, e.g. with a moisture above 80%, may degrade rapidly. Dried biomass
lends itself well to extraction since the drying process usually breaks down
cell walls,
allowing solvent to capture the molecules inside. The temperature of the oven
and the
drying time depend on how much moisture is in the raw psychedelic fungus, and
on the
quantity of raw psychedelic fungus.
[232] The solvent may be selected from a range of different solvents,
including
lower aliphatic alcohols (C=1, 2, 3 or 4), water, alcohol-water mixtures,
strong alkaline
buffers, and strong acidic buffers. A wide range of solvent to solid ratios
can be used.
Typically, a 1 to 50:1 solvent-solid ratio (L:kg) may be used for the
extraction. The
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amount of solvent used generally varies according to the weight of the raw
psychedelic
fungus.
[233] In step 110, as a result of adding the solvent and soaking the
biomass of
dried, raw psychedelic fungus in the solvent, essential elements or
psychoactive
alkaloids found in the biomass, dissolve into the solvent. The solvent may be
at a low
or high temperature, and pressure may be applied to the solvent. In some
embodiments the solvent is at room temperature. The optimal temperature of
extraction varies depending on the solvent type used for the process. However,
the
optimal temperature for extraction is in range of 5-95 C. The useful
temperature
range spans most of the liquid state of the solvent used, and upper and lower
limits are
determined by physical practicalities and limits of the available apparatus.
Still, the
temperature of the solvent may be outside of this range in other embodiments.
The
duration of the extraction is from 10 minutes to 12 hours, with or without
agitation.
Optimum duration is determined by experimentation and depends on the chosen
solvent and the strength of agitation in the extraction vessel.
[234] If pressure is applied, it may be in the range of 50 kPa-100 MPa
above
atmospheric (7-15000 psig). The lower limit of pressure is indicative of when
a benefit
is seen in the rate at which the psychoactive alkaloids dissolve in the
solvent, since the
increased pressure may increase the reaction kinetics of the dissolution of
the
psychoactive alkaloids into the solvent. The upper limit is determined by what
is
physically practical given the constraints of equipment to safely operate
under high
pressure. Nevertheless, other pressures may be used. Solvent composition,
particle
size, and extraction temperature may determine how much pressure needs to be
applied.
[235] In some embodiments, the biomass of the psychoactive organism is
reduced
to a particle size of 6 millimeters (mm) to 0.03 mm prior to contacting with
the solvent.
In some embodiments, prior to contacting with the solvent, the biomass of the
psychoactive organism is reduced to a particle size of about 0.03 mm to about
6 mm. In
some embodiments, prior to contacting with the solvent, the biomass of the
psychoactive organism is reduced to a particle size of about 6 mm to about 5
mm,
about 6 mm to about 4 mm, about 6 mm to about 3 mm, about 6 mm to about 2 mm,
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about 6 mm to about 1 mm, about 6 mm to about 0.5 mm, about 6 mm to about 0.1
mm, about 6 mm to about 0.05 mm, about 6 mm to about 0.03 mm, about 5 mm to
about 4 mm, about 5 mm to about 3 mm, about 5 mm to about 2 mm, about 5 mm to
about 1 mm, about 5 mm to about 0.5 mm, about 5 mm to about 0.1 mm, about 5 mm
to about 0.05 mm, about 5 mm to about 0.03 mm, about 4 mm to about 3 mm, about
4
mm to about 2 mm, about 4 mm to about 1 mm, about 4 mm to about 0.5 mm, about
4
mm to about 0.1 mm, about 4 mm to about 0.05 mm, about 4 mm to about 0.03 mm,
about 3 mm to about 2 mm, about 3 mm to about 1 mm, about 3 mm to about 0.5
mm,
about 3 mm to about 0.1 mm, about 3 mm to about 0.05 mm, about 3 mm to about
0.03
mm, about 2 mm to about 1 mm, about 2 mm to about 0.5 mm, about 2 mm to about
0.1 mm, about 2 mm to about 0.05 mm, about 2 mm to about 0.03 mm, about 1 mm
to
about 0.5 mm, about 1 mm to about 0.1 mm, about 1 mm to about 0.05 mm, about 1
mm to about 0.03 mm, about 0.5 mm to about 0.1 mm, about 0.5 mm to about 0.05
mm, about 0.5 mm to about 0.03 mm, about 0.1 mm to about 0.05 mm, about 0.1 mm
to about 0.03 mm, or about 0.05 mm to about 0.03 mm. In some embodiments,
prior to
contacting with the solvent, the biomass of the psychoactive organism is
reduced to a
particle size of about 6 mm, about 5 mm, about 4 mm, about 3 mm, about 2 mm,
about
1 mm, about 0.5 mm, about 0.1 mm, about 0.05 mm, or about 0.03 mm. In some
embodiments, prior to contacting with the solvent, the biomass of the
psychoactive
organism is reduced to a particle size of at least about 6 mm, about 5 mm,
about 4 mm,
about 3 mm, about 2 mm, about 1 mm, about 0.5 mm, about 0.1 mm, or about 0.05
mm. In some embodiments, prior to contacting with the solvent, the biomass of
the
psychoactive organism is reduced to a particle size of at most about 5 mm,
about 4
mm, about 3 mm, about 2 mm, about 1 mm, about 0.5 mm, about 0.1 mm, about 0.05
mm, or about 0.03 mm.
[236] In some embodiments, the biomass of the psychoactive organism is
reduced
to a particle size of 1 millimeters (mm) to 0.03 mm prior to contacting with
the solvent.
In some embodiments, prior to contacting with the solvent, the biomass of the
psychoactive organism is reduced to a particle size of about 0.03 mm to about
1 mm. In
some embodiments, prior to contacting with the solvent, the biomass of the
psychoactive organism is reduced to a particle size of about 1 mm to about 0.9
mm,
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about 1 mm to about 0.8 mm, about 1 mm to about 0.7 mm, about 1 mm to about
0.6
mm, about 1 mm to about 0.5 mm, about 1 mm to about 0.4 mm, about 1 mm to
about
0.3 mm, about 1 mm to about 0.2 mm, about 1 mm to about 0.1 mm, about 1 mm to
about 0.05 mm, about 1 mm to about 0.03 mm, about 0.9 mm to about 0.8 mm,
about
0.9 mm to about 0.7 mm, about 0.9 mm to about 0.6 mm, about 0.9 mm to about
0.5
mm, about 0.9 mm to about 0.4 mm, about 0.9 mm to about 0.3 mm, about 0.9 mm
to
about 0.2 mm, about 0.9 mm to about 0.1 mm, about 0.9 mm to about 0.05 mm,
about
0.9 mm to about 0.03 mm, about 0.8 mm to about 0.7 mm, about 0.8 mm to about
0.6
mm, about 0.8 mm to about 0.5 mm, about 0.8 mm to about 0.4 mm, about 0.8 mm
to
about 0.3 mm, about 0.8 mm to about 0.2 mm, about 0.8 mm to about 0.1 mm,
about
0.8 mm to about 0.05 mm, about 0.8 mm to about 0.03 mm, about 0.7 mm to about
0.6
mm, about 0.7 mm to about 0.5 mm, about 0.7 mm to about 0.4 mm, about 0.7 mm
to
about 0.3 mm, about 0.7 mm to about 0.2 mm, about 0.7 mm to about 0.1 mm,
about
0.7 mm to about 0.05 mm, about 0.7 mm to about 0.03 mm, about 0.6 mm to about
0.5
mm, about 0.6 mm to about 0.4 mm, about 0.6 mm to about 0.3 mm, about 0.6 mm
to
about 0.2 mm, about 0.6 mm to about 0.1 mm, about 0.6 mm to about 0.05 mm,
about
0.6 mm to about 0.03 mm, about 0.5 mm to about 0.4 mm, about 0.5 mm to about
0.3
mm, about 0.5 mm to about 0.2 mm, about 0.5 mm to about 0.1 mm, about 0.5 mm
to
about 0.05 mm, about 0.5 mm to about 0.03 mm, about 0.4 mm to about 0.3 mm,
about
0.4 mm to about 0.2 mm, about 0.4 mm to about 0.1 mm, about 0.4 mm to about
0.05
mm, about 0.4 mm to about 0.03 mm, about 0.3 mm to about 0.2 mm, about 0.3 mm
to
about 0.1 mm, about 0.3 mm to about 0.05 mm, about 0.3 mm to about 0.03 mm,
about
0.2 mm to about 0.1 mm, about 0.2 mm to about 0.05 mm, about 0.2 mm to about
0.03
mm, about 0.1 mm to about 0.05 mm, about 0.1 mm to about 0.03 mm, or about
0.05
mm to about 0.03 mm. In some embodiments, prior to contacting with the
solvent, the
biomass of the psychoactive organism is reduced to a particle size of about 1
mm,
about 0.9 mm, about 0.8 mm, about 0.7 mm, about 0.6 mm, about 0.5 mm, about
0.4
mm, about 0.3 mm, about 0.2 mm, about 0.1 mm, about 0.05 mm, or about 0.03 mm.
In
some embodiments, prior to contacting with the solvent, the biomass of the
psychoactive organism is reduced to a particle size of at least about 1 mm,
about 0.9
mm, about 0.8 mm, about 0.7 mm, about 0.6 mm, about 0.5 mm, about 0.4 mm,
about
Date Recue/Date Received 2022-07-25
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0.3 mm, about 0.2 mm, about 0.1 mm, or about 0.05 mm. In some embodiments,
prior
to contacting with the solvent, the biomass of the psychoactive organism is
reduced to a
particle size of at most about 0.9 mm, about 0.8 mm, about 0.7 mm, about 0.6
mm,
about 0.5 mm, about 0.4 mm, about 0.3 mm, about 0.2 mm, about 0.1 mm, about
0.05
mm, or about 0.03 mm.
[237] In some embodiments, the biomass of the psychoactive organism is
reduced
to a particle size of at least 0.074 mm prior to contacting with the solvent.
[238] The extraction results in an extraction slurry, which is formed of
undissolved
and insoluble solids from the mixture of biomass and solvent, which now
carries
dissolved extract. Some of the undissolved solids may be undesirable
components.
[239] In step 120, the extraction slurry is filtered, resulting in a
residue (i.e., the
undissolved portion of the biomass) and filtrate. The filtering step may be
carried out
with the extraction slurry still hot, or it may first be allowed to cool. The
extraction and
filtration steps may be repeated multiple times on the same residue, with a
fresh batch
of solvent, which may have the same composition as the first solvent, or it
may be a
different solvent.
[240] In step 130, if the filtrate results from using a strongly acidic or
alkaline
solvent, then the filtrate is brought closer to neutral, e.g., to a pH between
4 and 9 or
thereabouts. Desirable effects, such as more complete extraction, preservation
of the
alkaloids from decomposition, or the ability to selectively extract certain
specific
alkaloids, are seen during the extraction stage when stronger acids or alkalis
are used
compared to weaker ones.
[241] In step 140, evaporation of some or all of the solvent from the
filtrate results in
a concentrated slurry 150 (liquid and solids) or just solids. If the solvent
is methanol,
then all of it is evaporated to reduce the likelihood of toxicity. For other
solvents, only
some of the solvent needs to be evaporated. In the case where solids are
obtained
from the evaporation, water is added to the solids to form the concentrated
slurry 150.
The solids tend not to dissolve back into solution because they are less
soluble in
methanol and ethanol, for example, than water. Also, the solids may be less
soluble in
the colder water that is added back than the warmer or hotter water that is
used for the
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extraction. Another reason is saturation of the solution or that some of the
solids are
irreversibly precipitated.
[242] In step 160, standardization of the concentrated slurry takes place.
The aim is
to stabilize the extract by adding stabilizer (e.g., ascorbic acid and
silica), and then
titrating with a carrier (e.g., maltodextrin) to result in a known
concentration of
psychoactive alkaloids. The slurry is analyzed for dry mass concentration and
alkaloid
content. The liquid component of the concentrated slurry is first analyzed
using a loss-
on-drying analysis and high-performance liquid chromatography coupled with
diode
array detection or mass spectrometry to determine the alkaloid content.
Depending on
the determined alkaloid content, non-toxic carriers are added to the
concentrated slurry
to provide a desired ratio between the weight of alkaloid and weight of
carrier in the
concentrated slurry. The added carriers, blending agents, excipients, flow
aids, etc.,
that may be used include maltodextrin from corn, potato, or tapioca, for
example, gum
arabic, silicon dioxide, microcrystalline cellulose, ascorbic acid, sodium
benzoate,
sodium phosphate, sodium citrate, rice hulls, and rice. A combination of any
of these
carriers may be used.
[243] In step 170, the concentrated slurry is dried to remove the remaining
solvent
or water, resulting in a powdered psilocybin mushroom extract with a known
concentration by weight of psychoactive compound(s). The extract is a powdered
psilocybin mushroom extract that may have, for example, a total psychoactive
alkaloid
concentration of 0.1-10% by dry weight. Other compounds may be included in the
extract. These may be sugars, proteins, carbohydrates, and fats, and may make
up
about half of the extract. Step 170 is optional, as it may be the intention to
produce a
liquid extract instead of a powdered extract.
[244] In some embodiments, the extraction is performed at a temperature
ranging
from 5-95 C. In some embodiments, the extraction is performed at a
temperature
ranging from about 5 C to about 95 C. In some embodiments, the extraction is
performed at a temperature ranging from about 5 C to about 10 C, about 5 C
to about
20 C, about 5 C to about 30 C, about 5 C to about 40 C, about 5 C to
about 50 C,
about 5 C to about 60 C, about 5 C to about 70 C, about 5 C to about 80
C, about 5
C to about 90 C, about 5 C to about 95 C, about 10 C to about 20 C, about
10 C to
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about 30 C, about 10 C to about 40 C, about 10 C to about 50 C, about 10
C to about
60 C, about 10 C to about 70 C, about 10 C to about 80 C, about 10 C to
about 90 C,
about 10 C to about 95 C, about 20 C to about 30 C, about 20 C to about
40 C, about
20 C to about 50 C, about 20 C to about 60 C, about 20 C to about 70 C,
about 20 C
to about 80 C, about 20 C to about 90 C, about 20 C to about 95 C, about
30 C to
about 40 C, about 30 C to about 50 C, about 30 C to about 60 C, about 30
C to about
70 C, about 30 C to about 80 C, about 30 C to about 90 C, about 30 C to
about 95 C,
about 40 C to about 50 C, about 40 C to about 60 C, about 40 C to about
70 C, about
40 C to about 80 C, about 40 C to about 90 C, about 40 C to about 95 C,
about 50 C
to about 60 C, about 50 C to about 70 C, about 50 C to about 80 C, about
50 C to
about 90 C, about 50 C to about 95 C, about 60 C to about 70 C, about 60
C to about
80 C, about 60 C to about 90 C, about 60 C to about 95 C, about 70 C to
about 80 C,
about 70 C to about 90 C, about 70 C to about 95 C, about 80 C to about
90 C, about
80 C to about 95 C, or about 90 C to about 95 C. In some embodiments, the
extraction is performed at a temperature ranging from about 5 C, about 10 C,
about 20
C, about 30 C, about 40 C, about 50 C, about 60 C, about 70 C, about 80
C, about
90 C, or about 95 C. In some embodiments, the extraction is performed at a
temperature ranging from at least about 5 C, about 10 C, about 20 C, about
30 C,
about 40 C, about 50 C, about 60 C, about 70 C, about 80 C, or about 90
C. In some
embodiments, the extraction is performed at a temperature ranging from at most
about
C, about 20 C, about 30 C, about 40 C, about 50 C, about 60 C, about 70
C,
about 80 C, about 90 C, or about 95 C.
[245] In other embodiments, the extraction is performed at a temperature
ranging
from 50-75 C. In some embodiments, the extraction is performed at a
temperature
ranging from about 50 C to about 75 C. In some embodiments, the extraction
is
performed at a temperature ranging from about 50 C to about 55 C, about 50
C to
about 60 C, about 50 C to about 65 C, about 50 C to about 70 C, about 50
C to about
75 C, about 55 C to about 60 C, about 55 C to about 65 C, about 55 C to
about 70 C,
about 55 C to about 75 C, about 60 C to about 65 C, about 60 C to about
70 C, about
60 C to about 75 C, about 65 C to about 70 C, about 65 C to about 75 C,
or about 70
C to about 75 C. In some embodiments, the extraction is performed at a
temperature
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ranging from about 50 C, about 55 C, about 60 C, about 65 C, about 70 C,
or about
75 C. In some embodiments, the extraction is performed at a temperature
ranging from
at least about 50 C, about 55 C, about 60 C, about 65 C, or about 70 C.
In some
embodiments, the extraction is performed at a temperature ranging from at most
about
55 C, about 60 C, about 65 C, about 70 C, or about 75 C.
[246] In some embodiments, the extraction is performed for a time period
ranging
from 10-720 minutes. For most cases, a time below 10 min would result in a
mostly
incomplete yield, and above 720 min the extraction may be incomplete but would
be
continuing at a negligible rate. In some embodiments, the extraction is
performed for a
time period ranging from about 10 min to about 720 min. In some embodiments,
the
extraction is performed for a time period ranging from about 10 min to about
100 min,
about 10 min to about 200 min, about 10 min to about 300 min, about 10 min to
about
400 min, about 10 min to about 500 min, about 10 min to about 600 min, about
10 min
to about 720 min, about 100 min to about 200 min, about 100 min to about 300
min,
about 100 min to about 400 min, about 100 min to about 500 min, about 100 min
to
about 600 min, about 100 min to about 720 min, about 200 min to about 300 min,
about
200 min to about 400 min, about 200 min to about 500 min, about 200 min to
about 600
min, about 200 min to about 720 min, about 300 min to about 400 min, about 300
min
to about 500 min, about 300 min to about 600 min, about 300 min to about 720
min,
about 400 min to about 500 min, about 400 min to about 600 min, about 400 min
to
about 720 min, about 500 min to about 600 min, about 500 min to about 720 min,
or
about 600 min to about 720 min. In some embodiments, the extraction is
performed for
a time period ranging from about 10 min, about 100 min, about 200 min, about
300 min,
about 400 min, about 500 min, about 600 min, or about 720 min. In some
embodiments, the extraction is performed for a time period ranging from at
least about
min, about 100 min, about 200 min, about 300 min, about 400 min, about 500
min,
or about 600 min. In some embodiments, the extraction is performed for a time
period
ranging from at most about 100 min, about 200 min, about 300 min, about 400
min,
about 500 min, about 600 min, or about 720 min.
[247] In another embodiment, and more usually, the extraction is performed
for a
time period ranging from 30-240 minutes. In some embodiments, the extraction
is
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performed for a time period ranging from about 30 min to about 240 min. In
some
embodiments, the extraction is performed for a time period ranging from about
30 min
to about 60 min, about 30 min to about 90 min, about 30 min to about 120 min,
about
30 min to about 150 min, about 30 min to about 180 min, about 30 min to about
210
min, about 30 min to about 240 min, about 60 min to about 90 min, about 60 min
to
about 120 min, about 60 min to about 150 min, about 60 min to about 180 min,
about
60 min to about 210 min, about 60 min to about 240 min, about 90 min to about
120
min, about 90 min to about 150 min, about 90 min to about 180 min, about 90
min to
about 210 min, about 90 min to about 240 min, about 120 min to about 150 min,
about
120 min to about 180 min, about 120 min to about 210 min, about 120 min to
about 240
min, about 150 min to about 180 min, about 150 min to about 210 min, about 150
min
to about 240 min, about 180 min to about 210 min, about 180 min to about 240
min, or
about 210 min to about 240 min. In some embodiments, the extraction is
performed for
a time period ranging from about 30 min, about 60 min, about 90 min, about 120
min,
about 150 min, about 180 min, about 210 min, or about 240 min. In some
embodiments, the extraction is performed for a time period ranging from at
least about
30 min, about 60 min, about 90 min, about 120 min, about 150 min, about 180
min, or
about 210 min. In some embodiments, the extraction is performed for a time
period
ranging from at most about 60 min, about 90 min, about 120 min, about 150 min,
about
180 min, about 210 min, or about 240 min.
[248] In
some embodiments, the extraction is performed at a pressure ranging from
7 to 20,000 psi (50kPa-138MPa). In some embodiments, the extraction is
performed at
a pressure ranging from about 7 psi (50kPa) to about 2,000 psi (14MPa). In
some
embodiments, the extraction is performed at a pressure ranging from about 7
psi
(50kPa) to about 300 psi (2MPa), about 7 psi (50kPa) to about 600 psi (4MPa),
about
7 psi (50kPa) to about 900 psi (6MPa), about 7 psi (50kPa) to about 1,200 psi
(8MPa),
about 7 psi (50kPa) to about 1,500 psi (10MPa), about 7 psi (50kPa) to about
1,800 psi
(12MPa), about 7 psi (50kPa) to about 2,000 psi (14MPa), about 300 psi (2MPa)
to
about 600 psi (4MPa), about 300 psi (2MPa) to about 900 psi (6MPa), about 300
psi
(2MPa) to about 1,200 psi (8MPa), about 300 psi (2MPa) to about 1,500 psi
(10MPa),
about 300 psi (2MPa) to about 1,800 psi (12MPa), about 300 psi (2MPa) to about
2,000
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psi (14MPa), about 600 psi (4MPa) to about 900 psi (6MPa), about 600 psi
(4MPa) to
about 1,200 psi (8MPa), about 600 psi (4MPa) to about 1,500 psi (10MPa), about
600
psi (4MPa) to about 1,800 psi (12MPa), about 600 psi (4MPa) to about 2,000 psi
(14MPa), about 900 psi (6MPa) to about 1,200 psi (8MPa), about 900 psi (6MPa)
to
about 1,500 psi (10MPa), about 900 psi (6MPa) to about 1,800 psi (12MPa),
about 900
psi (6MPa) to about 2,000 psi (14MPa), about 1,200 psi (8MPa) to about 1,500
psi
(10MPa), about 1,200 psi (8MPa) to about 1,800 psi (12MPa), about 1,200 psi
(8MPa)
to about 2,000 psi (14MPa), about 1,500 psi (10MPa) to about 1,800 psi
(12MPa),
about 1,500 psi (10MPa) to about 2,000 psi (14MPa), or about 1,800 psi (12MPa)
to
about 2,000 psi (14MPa). In some embodiments, the extraction is performed at a
pressure ranging from about 7 psi (50kPa), about 300 psi (2MPa), about 600 psi
(4MPa), about 900 psi (6MPa), about 1,200 psi (8MPa), about 1,500 psi (10MPa),
about 1,800 psi (12MPa), or about 2,000 psi (14MPa). In some embodiments, the
extraction is performed at a pressure ranging from at least about 7 psi
(50kPa), about
300 psi (2MPa), about 600 psi (4MPa), about 900 psi (6MPa), about 1,200 psi
(8MPa),
about 1,500 psi (10MPa), or about 1,800 psi (12MPa). In some embodiments, the
extraction is performed at a pressure ranging from at most about 300 psi
(2MPa), about
600 psi (4MPa), about 900 psi (6MPa), about 1,200 psi (8MPa), about 1,500 psi
(10MPa), about 1,800 psi (12MPa), or about 2,000 psi (14MPa).
[249] In yet another embodiment, the extraction is performed at a pressure
ranging
from 10 to 20 psi (70-140kPa). In some embodiments, the extraction is
performed at a
pressure ranging from about 10 psi (70kPa) to about 20 psi (140kPa). In some
embodiments, the extraction is performed at a pressure ranging from about 10
psi
(70kPa) to about 12 psi (80kPa), about 10 psi (70kPa) to about 14 psi
(100kPa), about
psi (70kPa) to about 16 psi (110kPa), about 10 psi (70kPa) to about 18 psi
(120kPa),
about 10 psi (70kPa) to about 20 psi (140kPa), about 12 psi (80kPa) to about
14 psi
(100kPa), about 12 psi (80kPa) to about 16 psi(110kPa), about 12 psi (80kPa)
to about
18 psi (120kPa), about 12 psi (80kPa) to about 20 psi (140kPa), about 14 psi
(100kPa)
to about 16 psi (110kPa), about 14 psi (100kPa) to about 18 psi (120kPa),
about 14 psi
(100kPa) to about 20 psi (140kPa), about 16 psi (110kPa) to about 18 psi
(120kPa),
about 16 psi (110kPa) to about 20 psi (140kPa), or about 18 psi (120kPa) to
about 20
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psi (140kPa). In some embodiments, the extraction is performed at a pressure
ranging
from about 10 psi (70kPa), about 12 psi (80kPa), about 14 psi (100kPa), about
16 psi
(110kPa), about 18 psi (120kPa), or about 20 psi (140kPa). In some
embodiments, the
extraction is performed at a pressure ranging from at least about 10 psi
(70kPa), about
12 psi (80kPa), about 14 psi (100kPa), about 16 psi (110kPa), or about 18 psi
(120kPa). In some embodiments, the extraction is performed at a pressure
ranging from
at most about 12 psi (80kPa), about 14 psi (100kPa), about 16 psi (110Pa),
about 18
psi (120kPa), or about 20 psi (140kPa).
[250] In
some embodiments, the extraction is performed with a solvent to solid ratio
in the range 10 to 100 mL/g, wherein the solid is the dried powdered biomass.
In one
embodiment, the extraction is performed with a solvent to solid ratio of 20
mL/g. In
some embodiments, the extraction is performed with a solvent to solid ratio in
the range
of about 10 mL/g to about 100 mL/g. In some embodiments, the extraction is
performed
with a solvent to solid ratio in the range of about 10 mL/g to about 20 mL/g,
about 10
mL/g to about 30 mL/g, about 10 mL/g to about 40 mL/g, about 10 mL/g to about
50
mL/g, about 10 mL/g to about 60 mL/g, about 10 mL/g to about 70 mL/g, about 10
mL/g
to about 80 mL/g, about 10 mL/g to about 90 mL/g, about 10 mL/g to about 100
mL/g,
about 20 mL/g to about 30 mL/g, about 20 mL/g to about 40 mL/g, about 20 mL/g
to
about 50 mL/g, about 20 mL/g to about 60 mL/g, about 20 mL/g to about 70 mL/g,
about 20 mL/g to about 80 mL/g, about 20 mL/g to about 90 mL/g, about 20 mL/g
to
about 100 mL/g, about 30 mL/g to about 40 mL/g, about 30 mL/g to about 50
mL/g,
about 30 mL/g to about 60 mL/g, about 30 mL/g to about 70 mL/g, about 30 mL/g
to
about 80 mL/g, about 30 mL/g to about 90 mL/g, about 30 mL/g to about 100
mL/g,
about 40 mL/g to about 50 mL/g, about 40 mL/g to about 60 mL/g, about 40 mL/g
to
about 70 mL/g, about 40 mL/g to about 80 mL/g, about 40 mL/g to about 90 mL/g,
about 40 mL/g to about 100 mL/g, about 50 mL/g to about 60 mL/g, about 50 mL/g
to
about 70 mL/g, about 50 mL/g to about 80 mL/g, about 50 mL/g to about 90 mL/g,
about 50 mL/g to about 100 mL/g, about 60 mL/g to about 70 mL/g, about 60 mL/g
to
about 80 mL/g, about 60 mL/g to about 90 mL/g, about 60 mL/g to about 100
mL/g,
about 70 mL/g to about 80 mL/g, about 70 mL/g to about 90 mL/g, about 70 mL/g
to
about 100 mL/g, about 80 mL/g to about 90 mL/g, about 80 mL/g to about 100
mL/g, or
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about 90 mL/g to about 100 mL/g. In some embodiments, the extraction is
performed
with a solvent to solid ratio in the range of about 10 mL/g, about 20 mL/g,
about 30
mL/g, about 40 mL/g, about 50 mL/g, about 60 mL/g, about 70 mL/g, about 80
mL/g,
about 90 mL/g, or about 100 mL/g. In some embodiments, the extraction is
performed
with a solvent to solid ratio in the range of at least about 10 mL/g, about 20
mL/g, about
30 mL/g, about 40 mL/g, about 50 mL/g, about 60 mL/g, about 70 mL/g, about 80
mL/g,
or about 90 mL/g. In some embodiments, the extraction is performed with a
solvent to
solid ratio in the range of at most about 20 mL/g, about 30 mL/g, about 40
mL/g, about
50 mL/g, about 60 mL/g, about 70 mL/g, about 80 mL/g, about 90 mL/g, or about
100
mL/g.
Extraction Process 2
[251] In some embodiments, referring to FIG. 15, a process for extracting
the
psychoactive alkaloid is shown. The psychoactive alkaloid source is dried in
step 1500
by techniques known in the art, such as using a forced air oven.
[252] In step 1510, the dried psychoactive alkaloid source or dried biomass
is mixed
with a solvent and/or left to soak. The solvent in which the extract is
carried or
dissolved may be a primary aliphatic alcohol, a ketone, water, and any
combination
selected therefrom. In some embodiments, the primary aliphatic alcohol is a C1-
4
alcohol. In some embodiments, the primary aliphatic alcohol is 5% ethanol. In
some
embodiments, the primary aliphatic alcohol is ethanol. In some embodiments,
the
ketone is a C3-4 ketone. In yet other embodiments, the water is selected from
deionized, distilled, reverse osmosis, or otherwise purified water, which is
substantially
without free ions. In other embodiments, the water is not purified. In an
exemplary
embodiment, the solvent is a hydro-ethanol mixture with 3 parts of ethanol to
1 part of
water, by weight.
[253] In step 1520, the mixture of the solvent and the dried psychoactive
alkaloid
source is filtered to obtain a filtrate and a filtrate residue. Some or all of
the solvent is
evaporated from the filtrate residue to obtain the psychoactive alkaloid
extract.
Standardization of the obtained psychoactive alkaloid extract is carried out
according to
the standardization step 160described above.
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[254] Optionally, the filtrate residue obtained in step 1520 is extracted
with the
solvent again. The filtrate residue is extracted by repeating the steps 1510
and 1520.
This results in another filtrate. The first filtrate and the second filtrate
are mixed
together after their respective filtration steps to result in a bulk filtrate.
The solvent from
this bulk filtrate is partially evaporated to obtain the psychoactive alkaloid
extract in a
slurry form for the standardization step 160.
[255] In one embodiment, the extraction is carried out at a temperature
ranging from
5-95 C. The useful temperature range spans most of the liquid state of the
solvent
used, and upper and lower limits are determined by physical practicalities and
limits of
the available apparatus. Still, the temperature of the solvent may be outside
of this
range in other embodiments.
[256] In other embodiments, the extraction is carried out at a temperature
of 70 C.
Temperature, and pressure if applied, are generally selected so that the
solvent does
not boil if elevated temperatures are used. In one embodiment, the extraction
is carried
out for a time duration ranging from 10 minutes to 12 hours. In yet another
embodiment, the extraction is carried out for a time duration of 4 hours.
Extraction Process 3
[257] In FIG. 10, the extracting step 1000 may include, as an example,
extracting
psychoactive alkaloids from raw, psychedelic mushrooms. The mushrooms are
dried
and ground to result in a dried biomass. The next step involves heating the
dried
biomass in a solvent in order for the extraction to occur. The obtained slurry
is filtered
to obtain a first filtrate and a first residue. The first residue undergoes a
second
extraction, using a second solvent to obtain a second slurry, which is then
filtered to
obtain a second filtrate and a second residue. The first filtrate and the
second filtrate
are mixed to obtain the psychoactive alkaloid extract. More extract can be
obtained this
way, i.e., by splitting the solvent into two or more batches and using each
one
sequentially to soak the biomass, compared to using a single volume of
solvent.
[258] The extraction may further include completely or partially
concentrating the
obtained psychoactive alkaloid extract, by evaporation of the solvent from the
combined
filtrates.
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[259] In one embodiment, the first solvent and the second solvent are
selected from
a primary aliphatic alcohol, a ketone, water, and any combination therefrom.
In one
embodiment, the primary aliphatic alcohol is a C1-4 alcohol. In one
embodiment, the
ketone is a 03-4 ketone. In another embodiment, the first solvent is an
ethanol-water
mixture with 3 parts ethanol to 1 part water by weight. In another embodiment,
the
second solvent is an ethanol-water mixture with 3 parts ethanol to 1 part
water by
weight. In yet another embodiment, the water is selected from deionized,
distilled,
reverse osmosis, or otherwise purified water, which has substantially no free
ions. The
selection of the solvent will depend on the nature of the starting material
for extraction
and the reaction conditions, according to which a person of skill in the art
can make the
appropriate solvent selection.
[260] In one embodiment, the extraction is carried out at a temperature
ranging from
5-95 C. The useful temperature range spans most of the liquid state of the
solvent
used, and upper and lower limits are determined by physical practicalities and
limits of
the available apparatus. Still, the temperature of the solvent may be outside
of this
range in other embodiments.
[261] In another embodiment, the extraction is carried out at a temperature
of 70 C.
Temperature and pressure, if applied, are generally selected so that the
solvent does
not boil if elevated temperatures are used. In one embodiment, the extraction
is carried
out for a time duration ranging from 10 minutes to 12 hours. In yet another
embodiment, the extraction is carried out for a time duration of 4 hours.
Extraction Process 4
[262] In one embodiment, referring to FIG. 18, a basic process for
obtaining a
psychoactive alkaloid extract with a desired amount of a phosphorylated
psychoactive
alkaloid and a desired amount of a dephosphorylated psychoactive alkaloid is
shown.
The phosphorylated alkaloid may be psilocybin, baeocystin, norbaeocystin,
aeruginascin, or any combination selected therefrom. The dephosphorylated
alkaloid
may be psilocin, norpsilocin, 4-hydroxytryptamine, N,N,N-trimethy1-4-
hydroxytryptamine, or any combination selected therefrom. The control aspect
of the
present invention relates to psychoactive alkaloids that have phosphorylated
forms and
not to other psychoactive alkaloids that may be present in a psychoactive
alkaloid
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source. Depending on the strain or harvest, there may be no or substantially
no
phosphorylatable psychoactive alkaloids in the psychoactive alkaloid source,
or they
may represent as much 80-90% of the total alkaloid content.
[263] The process includes step 1800 of obtaining powdered biomass from a
psychoactive alkaloid source. The powdered biomass is obtained by drying and
pulverizing a psychoactive alkaloid source. The drying is carried out via
vacuum
desiccation, freeze drying, timed forced air drying, or other existing drying
method, to
obtain a dried biomass. The pulverization is carried out by milling, grinding,
or other
method to reduce the particle size of the dried biomass. In one embodiment,
the drying
is carried out in a forced air oven completely shielded from all light at 20-
30 C for a
time period of 5-10 hours. However, there is room for optimization of the
drying step,
using different temperatures (e.g., 10-50 C) and different durations.
[264] In one embodiment, the drying is carried out in a manner to not
promote the
conversion of phosphorylated psychoactive alkaloid. Taking care not to bruise
while
harvesting the psychoactive alkaloid source, harvesting at the right time of
the fruiting
body life cycle, potentially freeze-drying the fruiting body, low heat
desiccation, and
gentle air drying are all ways that reduce the conversion of phosphorylated
psychoactive alkaloid to dephosphorylated psychoactive alkaloid. It may be
feasible to
harvest the fungi in a basic or methanol environment or soak the whole
mycelium in
methanol and cut the fruits while soaked. Once dried, there is negligible
conversion, so
that subsequent pulverization has little effect on it, unless moisture is
added back. The
prevention of conversion of phosphorylated psychoactive alkaloid to
dephosphorylated
psychoactive alkaloid during harvesting allows for the preparation of
psychoactive
alkaloid extracts, by the present process, having a total phosphorylatable
psychoactive
alkaloid content that is up to 100% by weight of phosphorylated psychoactive
alkaloid.
[265] Step 1810 involves extracting a psychoactive alkaloid from the dried
powdered biomass with an acidified solvent or a basified solvent to obtain a
psychoactive alkaloid liquid with a specific pH, wherein the specific pH is
lower than 3.5
or greater than 10.5. Between pH 3.5 and pH 10.5, the conditions are such that
psilocybin is readily converted to psilocin, and psilocin is converted to the
quinoid dimer,
which is completely inactive.
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[266] When used, the acid may be acetic acid, adipic acid, ascorbic acid,
phosphoric acid, ammonium aluminum sulphate, ammonium citrate dibasic,
ammonium
citrate monobasic, calcium citrate, calcium fumarate, calcium gluconate,
calcium
phosphate dibasic, calcium phosphate monobasic, hydrochloric acid, sulfuric
acid
monobasic, calcium phosphate tribasic, citric acid, fumaric acid, gluconic
acid,
magnesium fumarate, malic acid, phosphoric acid, potassium acid tartrate,
potassium
citrate, potassium fumarate, sodium citrate, sodium fumarate, sodium
gluconate,
sodium lactate, sodium potassium hexametaphosphate, sodium potassium tartrate,
sodium potassium tripolyphosphate, sodium pyrophosphate tetrabasic, sodium
tripolyphosphate, tartaric acid, and any combination of one or more of these.
In some
embodiments, the acid is either only hydrochloric acid or only phosphoric
acid, for
example. It is also envisaged that other acids may be used, for example, non-
food-
grade acids that may be used by pharmaceuticals.
[267] When used, the base may be ammonium bicarbonate, ammonium carbonate,
ammonium hydroxide, calcium acetate, calcium carbonate, calcium chloride,
calcium
hydroxide, calcium lactate, calcium oxide, calcium phosphate dibasic, calcium
phosphate monobasic, magnesium carbonate, potassium aluminum sulphate,
potassium bicarbonate, potassium carbonate, potassium hydroxide, potassium
lactate,
potassium phosphate dibasic, potassium pyrophosphate tetrabasic, potassium
phosphate tribasic, potassium tripolyphosphate, sodium acetate, sodium acid
pyrophosphate, sodium aluminum phosphate, sodium aluminum sulphate, sodium
bicarbonate, sodium bisulphate, sodium carbonate, sodium hexametaphosphate,
sodium hydroxide, sodium lactate, sodium phosphate dibasic, sodium phosphate
monobasic, sodium phosphate tribasic, or any combination therefrom. In one
embodiment, the base is solely sodium hydroxide, for example. Other bases may
be
used in other embodiments, for example, non-food-grade bases that may be used
by
pharmaceuticals.
[268] After adding the acidified solvent or the basified solvent, the
psychoactive
alkaloid liquid has a pH ranging from 0.5 to 3.5 or from 10.5 to 13.5,
respectively. In an
exemplary embodiment, the pH of the psychoactive alkaloid liquid obtained
after
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addition of the basified solvent is 13. In another exemplary embodiment, the
pH of the
psychoactive alkaloid liquid obtained after addition of the acidified solvent
is 2.
[269] The pH is adjusted in the extraction step 1810 to halt or promote
conversion
of phosphorylated psychoactive alkaloid to dephosphorylated psychoactive
alkaloid,
thus allowing the preparation of the psychoactive alkaloid liquid with the
desired amount
of a phosphorylated psychoactive alkaloid and a desired amount of a
dephosphorylated
psychoactive alkaloid. A specific pH lower than 3.5 promotes the conversion of
the
phosphorylated psychoactive alkaloid to the dephosphorylated psychoactive
alkaloid. A
specific pH greater than 10.5 halts the conversion of the phosphorylated
psychoactive
alkaloid to the dephosphorylated psychoactive alkaloid.
[270] In step 1820 of the process, the pH of the obtained psychoactive
alkaloid
liquid is adjusted to a pH ranging from 3.5 to 4.5. The pH is adjusted by
adding a base
or an acid. The pH is adjusted to a value in this range as the psychoactive
alkaloid
liquid exhibits a good anti-microbial stability in this pH range. Also, there
is no
dephosphorylation at this pH after the alkaloids are removed from the biomass,
which
points to enzymatic hydrolysis being responsible for conversion in the source
of the
psychoactive alkaloids. In exemplary embodiments, the base is sodium
hydroxide, and
the acid is citric acid. Any other appropriate acid or base can be used to
adjust the pH,
which a person of skill in the art may determine. The selection of the acid or
the base
will depend upon the nature of the pH of the psychoactive alkaloid liquid
prior to
adjusting it to the range of 3.5-4.5, according to which a person of skill in
the art can
make the appropriate acid or base selection.
[271] Step 1830 of the process involves evaporating the solvent from the
psychoactive alkaloid solution to obtain the psychoactive alkaloid extract
with the
desired amount of the phosphorylated psychoactive alkaloid and the desired
amount of
the dephosphorylated psychoactive alkaloid. The solvent is completely or
partially
evaporated to result in the psychoactive alkaloid extract (slurry or powder)
with the
desired amount of the phosphorylated psychoactive alkaloid and the desired
amount of
the dephosphorylated psychoactive alkaloid. The evaporation is carried out by
methods such as air drying, rotary evaporation, or other existing methods to
evaporate
solvent from psychoactive alkaloid liquid. At this point in time, away from
the biomass,
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psilocybin and/or psilocin are fairly heat resistant, more so under vacuum,
thus, rotary
evaporation, for example, is a suitable process. The desired amount of the
phosphorylated psychoactive alkaloid is 0-100% of a phosphorylatable
psychoactive
alkaloid content in the psychoactive alkaloid extract. The desired amount of
the
dephosphorylated psychoactive alkaloid is the remainder of the total
phosphorylatable
psychoactive alkaloid content in the psychoactive alkaloid extract.
[272] In some embodiments, when the psychoactive alkaloid liquid has a pH
greater
than 10.5 during the extraction step, the desired amount of the phosphorylated
psychoactive alkaloid is 50-90% by weight (crude extraction) of the total
phosphorylatable psychoactive alkaloid content in the psychoactive alkaloid
extract,
without going to onerous lengths in selecting the raw material. The desired
amount of
the dephosphorylated psychoactive alkaloid is the remainder of the total
phosphorylatable psychoactive alkaloid content in the psychoactive alkaloid
extract.
[273] In another embodiment, when using a psychoactive alkaloid source that
has
not undergone conversion (i.e. no significant conversion) of any
phosphorylated
alkaloid to dephosphorylated alkaloid, and when the psychoactive alkaloid
liquid has a
pH greater than 10.5 during the extraction step, the desired amount of the
phosphorylated psychoactive alkaloid is 100% by weight of the total
phosphorylatable
psychoactive alkaloid content in the psychoactive alkaloid extract. The
desired amount
of the dephosphorylated psychoactive alkaloid is 0% by weight of the total
phosphorylatable psychoactive alkaloid content in the psychoactive alkaloid
extract.
[274] In yet another embodiment, when the psychoactive alkaloid liquid has
a pH
lower than 3.5 during the extraction step, the desired amount of the
phosphorylated
psychoactive alkaloid is 0% by weight of the total phosphorylatable
psychoactive
alkaloid content in the psychoactive alkaloid extract. The desired amount of
the
dephosphorylated psychoactive alkaloid is 100% by weight of the total
phosphorylatable psychoactive alkaloid content in the psychoactive alkaloid
extract.
Even with neutral hydro-ethanol extraction, a large portion of psilocybin may
be
converted to psilocin. However, the low pH environment (<3.5) protects the
psilocin
from oxidation.
Extraction Process 5
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[275] In some embodiments, referring to FIG. 19, additional, optional steps
in the
extraction step 1810 are shown. Step 1900 is performed by adding the acidified
solvent
or the basified solvent to the powered biomass. The obtained psychoactive
liquid has a
specific pH, the specific pH being lower than 3.5 or greater than 10.5. After
the addition
of the acidified solvent or the basified solvent, the powered biomass and the
solvent are
mixed, followed by step 1910 of filtration to result in the extracted filtrate
of step 1920
(i.e. psychoactive alkaloid liquid). To this obtained filtrate, the acid or
the base of step
1820 is added to adjust the pH to within the range of 3.5-4.5.
[276] In some embodiments, the extraction step comprises further extracting
the
psychoactive alkaloid by repeating the extraction step. Filtrate residue from
step 1910
is collected, and to this filtrate residue, the same or a different acidified
solvent or the
same or a different basified solvent is added. The resulting mixture is mixed
followed
by filtration to obtain another filtrate. This filtrate and the previous
filtrate are mixed
together to result in a bulk filtrate. To this bulk filtrate the acid or the
base is added to
adjust the pH to 3.5-4.5 according to step 1820.
[277] In some embodiments, further extraction of the filtrate obtained
after
extraction with the acidified or the basified solvent is repeated until a
required amount of
the phosphorylated psychoactive alkaloid and/or the dephosphorylated
psychoactive
alkaloid is extracted. The number of extraction cycles to be repeated may
depend on
various variable factors such as the source of the psychoactive alkaloid and
the
solubility of the psychoactive alkaloid in the acidified or the basified
solvent.
[278] The solvent in the evaporation step can be completely or partially
evaporated,
to result in a powdered solid or a slurry. Evaporation may be paused, for
standardization, and continued after.
Extraction Process 6
[279] Referring to FIG. 20, the present invention also relates to a process
for
obtaining a psychoactive alkaloid composition with a psychoactive alkaloid
extract and
one or more excipients.
[280] At step 2000 the evaporation step 1830 (FIG. 18) is paused when a
portion of
the solvent has been evaporated from the psychoactive alkaloid liquid to
obtain a
psychoactive alkaloid slurry. The evaporation of a portion of the solvent,
before
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collection of the psychoactive alkaloid slurry for standardization, is done to
obtain a
quantity of a psychoactive alkaloid slurry that is easy to handle in the
subsequent steps
of the standardization process. The quantity of the portion of the solvent to
be
evaporated before pausing the evaporation is not so much as to make it too
viscous to
handle well. The quantity of the portion of the solvent to be evaporated will
depend on
various factors, for example, but not limited to, the contents of the
psychoactive alkaloid
liquid and the quantity of the psychoactive alkaloid liquid present at the
beginning of the
evaporation step.
[281] Step 2010 includes standardizing the obtained psychoactive alkaloid
slurry by
adding thereto a measured quantity of one or more excipients to obtain a
standardized
slurry with a specific amount of psychoactive alkaloid content. The specified
amount of
the total psychoactive alkaloid content in the psychoactive alkaloid
composition is
achieved by first determining the proportion, by weight, of solids in the
psychoactive
alkaloid slurry. The weight proportion of the psychoactive alkaloids in the
slurry is also
determined. Then, a measured amount of one or more excipients is added to a
measured amount of the psychoactive alkaloid slurry. Thus, after evaporation
of the
remaining solvent, the resultant composition is a standardized composition
with a
specified total psychoactive alkaloid content.
[282] Step 2020 includes continuing the evaporating step 1830 (FIG. 18) by
drying
the standardized slurry to obtain a psychoactive alkaloid composition with the
psychoactive alkaloid extract and the one or more excipients.
[283] The evaporation in the standardization process is carried out by
methods such
as air drying, rotary evaporation, or other methods known in the art to
suitably
evaporate solvent from psychoactive alkaloid slurry.
[284] The psychoactive alkaloid composition obtained has a specific total
psychoactive alkaloid content in the composition due to the standardizing
step. The
specified amount of the total psychoactive alkaloid content may be accurate to
one or
two decimal places, or one or two significant figures depending on how
accurately the
measurements are made during the standardization process.
[285] Thus, the psychoactive alkaloid composition obtained has a specific
amount
of a total psychoactive alkaloid content, which is made up of a desired amount
of
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phosphorylated psychoactive alkaloid and a desired amount of dephosphorylated
psychoactive alkaloid, and possibly other psychoactive alkaloids that are not
phosphorylatable.
[286] In one embodiment, the specific amount of the total psychoactive
alkaloid
content in the psychoactive alkaloid composition ranges from 0.1-99% by weight
of the
composition, the higher concentrations being obtained when purifying steps are
included. Purifying may involve treating the extract with a resinous material
prior to the
standardization step, to remove impurities such as sugars and proteins. In
another
embodiment, the specific amount of the total psychoactive alkaloid content in
the
psychoactive alkaloid composition ranges from 0.1-10% by weight of the
composition,
which may be achieved without purifying steps. In an exemplary embodiment, the
specific amount of the total psychoactive alkaloid content in the psychoactive
alkaloid
composition is 0.53% by weight of the composition. In yet another exemplary
embodiment, the specific amount of the total psychoactive alkaloid content in
the
psychoactive alkaloid composition is 0.501% by weight of the composition.
[287] In some embodiments, the desired amount of the phosphorylated
psychoactive alkaloid is 0-100% by weight of a total phosphorylatable
psychoactive
alkaloid content in the psychoactive alkaloid extract, and the desired amount
of the
dephosphorylated psychoactive alkaloid is the remainder of the total
phosphorylatable
psychoactive alkaloid content in the psychoactive alkaloid extract. In one
embodiment,
the desired amount of the phosphorylated psychoactive alkaloid is 10-90% by
weight of
the total phosphorylatable psychoactive alkaloid content in the psychoactive
alkaloid
extract, and the desired amount of the dephosphorylated psychoactive alkaloid
is the
remainder of the total phosphorylatable psychoactive alkaloid content in the
psychoactive alkaloid extract. In another embodiment, the desired amount of
the
phosphorylated psychoactive alkaloid is 0% by weight of the total
phosphorylatable
psychoactive alkaloid content in the psychoactive alkaloid extract, and the
desired
amount of the dephosphorylated psychoactive alkaloid is 100% by weight of the
total
phosphorylatable psychoactive alkaloid content in the psychoactive alkaloid
extract. In
yet another embodiment, the desired amount of the phosphorylated psychoactive
alkaloid is 100% by weight of the total phosphorylatable psychoactive alkaloid
content
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in the psychoactive alkaloid extract, and the desired amount of the
dephosphorylated
psychoactive alkaloid is 0% by weight of the total phosphorylatable
psychoactive
alkaloid content in the psychoactive alkaloid extract.
Further Processes
[288] Referring to FIG. 10 additional, optional steps are shown well as the
basic
steps in the process. In one embodiment, the extraction step 1000 is followed
by
completely or partially concentrating the obtained psychoactive alkaloid
extract (or
solution) by evaporation of the solvent from the extract in step 1010. In
other
embodiments, step 1010 of partially or completely evaporating the solvent may
be
considered to be a part of the extraction step 1000. If the solvent from the
extract has
been completely evaporated in step 1010, then reverse osmosis water, more
solvent, or
another solvent is added back.
[289] In some embodiments, the process includes adding, in step 1020, an
acid or a
base to the psychoactive alkaloid extract obtained in step 1000 to obtain a
psychoactive
alkaloid solution with a specific pH.
[290] When used, the acid may be acetic acid, adipic acid, ascorbic acid,
phosphoric acid, ammonium aluminum sulphate, ammonium citrate dibasic,
ammonium
citrate monobasic, calcium citrate, calcium fumarate, calcium gluconate,
calcium
phosphate dibasic, calcium phosphate monobasic, hydrochloric acid, sulfuric
acid
monobasic, calcium phosphate tribasic, citric acid, fumaric acid, gluconic
acid,
magnesium fumarate, malic acid, phosphoric acid, potassium acid tartrate,
potassium
citrate, potassium fumarate, sodium citrate, sodium fumarate, sodium
gluconate,
sodium lactate, sodium potassium hexametaphosphate, sodium potassium tartrate,
sodium potassium tripolyphosphate, sodium pyrophosphate tetrabasic, sodium
tripolyphosphate, tartaric acid, and any combination of one or more of these.
In some
embodiments, the acid is either only hydrochloric acid or only phosphoric
acid, for
example. It is also envisaged that other acids may be used.
[291] When used, the base may be ammonium bicarbonate, ammonium carbonate,
ammonium hydroxide, calcium acetate, calcium carbonate, calcium chloride,
calcium
hydroxide, calcium lactate, calcium oxide, calcium phosphate dibasic, calcium
phosphate monobasic, magnesium carbonate, potassium aluminum sulphate,
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potassium bicarbonate, potassium carbonate, potassium hydroxide, potassium
lactate,
potassium phosphate dibasic, potassium pyrophosphate tetrabasic, potassium
phosphate tribasic, potassium tripolyphosphate, sodium acetate, sodium acid
pyrophosphate, sodium aluminum phosphate, sodium aluminum sulphate, sodium
bicarbonate, sodium bisulphate, sodium carbonate, sodium hexametaphosphate,
sodium hydroxide, sodium lactate, sodium phosphate dibasic, sodium phosphate
monobasic, sodium phosphate tribasic, or any combination therefrom. In one
embodiment, the base is solely sodium hydroxide, for example. Other bases may
be
used in other embodiments.
[292] In one embodiment, the specific pH psychoactive alkaloid solution has
a pH
ranging from 2.5 to 4.5, or from 9 to 10. In other embodiments, the specific
pH
psychoactive alkaloid solution has a pH of 3, 4, or 9.5. The selection of the
pH is
chosen in a manner to allow for the efficient adsorption of the psychoactive
alkaloids
onto the resin(s).
[293] In one embodiment, the process includes adding phosphoric acid to the
psychoactive alkaloid extract to achieve a pH of 4. In another embodiment, the
process
includes adding hydrochloric acid to the psychoactive alkaloid extract to
achieve a pH of
3. In yet another embodiment, the process includes adding sodium hydroxide to
the
psychoactive alkaloid extract to achieve a pH of 9.5.
[294] The process includes, in step 1030, optionally filtering,
centrifuging, or
clarifying the psychoactive alkaloid solution or specific pH psychoactive
alkaloid
solution, as the case may be, and utilizing the obtained filtrate for the next
step 1040 of
adsorption. Clarifying may be performed, for example, by adding an
agglomeration
agent.
[295] In step 1040, the process involves adsorbing the psychoactive
alkaloid(s) in
the solution onto the resin to obtain an adsorbed psychoactive alkaloid.
[296] In step 1050, the process involves washing the resin to remove
adsorbed
impurities from the resin. While not all the impurities are adsorbed onto the
resin, some
of them may be. The washing step, substantially, does not remove the adsorbed
psychoactive alkaloids. The washing solvent may be 100% ethanol, 100% reverse
osmosis water, or any other washing solvent used in ion-exchange resin
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chromatography, provided that the washing removes impurities or by-products
that are
adsorbed on the resin. Impurities or by-products may include, for example,
sugars,
carbohydrates, chitin, chitosan, fats, minerals, waxes, or proteins. There may
be one,
two, or more washing steps depending on the embodiment, and the same or
different
washing solvents may be used for each wash. In other embodiments, the
solvent(s) for
washing may include a primary aliphatic alcohol, a ketone, water, and any
combination
therefrom. In one embodiment, the primary aliphatic alcohol is a C1-4 alcohol.
In one
embodiment, the primary aliphatic alcohol is 5% ethanol. In one embodiment,
the
primary aliphatic alcohol is ethanol. In one embodiment, the ketone is a 03-4
ketone.
In yet another embodiment, the water is selected from deionized, distilled,
reverse
osmosis, or otherwise purified water that is substantially without free ions.
[297] After the washing, the process involves eluting, in step 1060, the
adsorbed
psychoactive alkaloid from the resin using a solvent to obtain a purified
psychoactive
alkaloid solution. The solvent may be an organic solvent, an acid, a base, or
water, a
combination of an organic solvent and a base, or a combination of an organic
solvent
and an acid, a combination of an organic solvent and water, a combination of
water and
a base, or combination of water and an acid. The result of the elution step is
a purified
psychoactive alkaloid solution.
[298] Following the elution, a further washing step 1070 may be employed
using
100% ethanol, for example, to wash the resin. This may be considered to be a
cleaning
step, to refresh the resin and make it ready to be used again in a subsequent
step or in
another process. In other embodiments, the solvent for further washing may be
a
primary aliphatic alcohol, a ketone, water, and any combination therefrom. In
one
embodiment, the primary aliphatic alcohol is a 01-4 alcohol. In one
embodiment, the
primary aliphatic alcohol is 5% ethanol. In one embodiment, the ketone is a 03-
4
ketone. In yet another embodiment, the water is selected from deionized,
distilled,
reverse osmosis, or otherwise purified water that is substantially without
free ions.
[299] The result of the elution is a purified psychoactive alkaloid
solution. In one
embodiment, the purified psychoactive alkaloid solution has a concentration of
0.07%
by weight of a psychoactive alkaloid, which is the concentration before
removal of some
or all of the solvent. This concentration may be different in other
embodiments,
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depending on the amount solvent used for the elution and the potency of the
raw
materials. In one embodiment, the purified psychoactive alkaloid solution is
concentrated by evaporating the solvent to form a purified psychoactive slurry
that has
at least of 5% by weight or more of a psychoactive alkaloid. In another
embodiment,
the purified psychoactive alkaloid slurry has 5-68% by weight of a
psychoactive
alkaloid. In yet other embodiments, the purified psychoactive alkaloid slurry
has a
concentration of psychoactive alkaloid outside these ranges, and, when dried,
can be
as low as 0.1% or as high as 99% dry wt/wW0.
[300] Optionally, the obtained purified psychoactive alkaloid solution is
further
purified by filtering the obtained purified psychoactive alkaloid solution to
obtain a
filtrate, and then repeating at least steps 1040 and 1060 with the obtained
filtrate.
[301] Different processes may employ the steps in a different order, and
some of
the steps may be repeated with the same or different parameters. For example,
in one
embodiment, starting from a solution of dissolved extract, the order of the
steps may be
1020, 1040, 1030, 1050, 1060, 1020, 1030, 1010, 1030, 1040, 1050 and 1060.
Variations
[302] Other water may be used in place of reverse osmosis water, which may
be
selected for its purity.
[303] Water purified by other purification technologies may be used instead
of
reverse osmosis water. In alternative embodiments, the solvent is 0.02% to
1.5%
acetic acid in water. In alternate embodiment, the solvent comprises 75%
ethanol, 25%
water, and 0.1M sodium hydroxide. In alternative embodiments the solvent is a
hydro-
methanol mixture, with a methanol content in the range of below 1% to 100%.
The
hydro-methanol based extraction follows the same steps as the extraction with
a
mixture of ethanol and water (FIG. 3) and may use lower soaking temperatures
due to
the lower boiling point of methanol. Also, the methanol/water mixture can be
evaporated to dryness instead of the partial evaporation in step 360, for
safety. If
evaporated to dryness, the extract is then formed by adding to the residual
solid. If not
evaporated to dryness, the residual slurry is diluted, if necessary for ease
of handling,
by adding reverse osmosis water to form the slurry. If not diluted, the
residual slurry is
used as the concentrated slurry. The result of evaporating the methanol is a
residue
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that is either solid or a slurry. Furthermore, the hydro-methanol solvent may
be
buffered with a strong acid or a strong alkali, following the processes in
FIGS. 6 and 7.
Again, however, the solvent may be completely evaporated instead of partially
(612,
712) in order to fully remove the methanol, with reverse osmosis water being
added to
the solid to form the concentrated slurry. If the solvent is not completely
evaporated, it
should be evaporated enough to remove all the methanol and leave a residual
slurry.
The residual slurry may optionally then be diluted, for ease of handling, with
reverse
osmosis water to form the concentrated slurry. If not diluted, the residual
slurry is used
as the concentrated slurry.
[304] The solvent may also be propan-1-ol, propan-2-ol, a butanol isomer,
or a
mixture of any or all of these with water, in any percentage ratio.
[305] Any of the solvents described herein may be used with any of the
mushroom
varieties that include psychoactive alkaloids.
[306] The temperature of extraction may be lowered to reduce conversion,
and the
duration may be in the range of 30 to 240 minutes.
[307] In other embodiments, other drying techniques, temperatures and
durations
may be used. It is possible in other embodiments to grind the dried biomass to
lower or
higher particle size than 200 mesh. For example, grinding to a mesh size of 40
would
work in some embodiments. The choice of solvent may have an impact on which
mesh
size to grind the dried mushrooms to. Note that, in other embodiments, the
grinding
step 614 may take place before or after the drying step 613.
[308] In general, unless otherwise indicated, singular elements may be in
the plural
and vice versa with no loss of generality.
[309] Temperatures that have been given to the nearest degree include all
temperatures within a range of 0.5 C of the given value. Likewise, numbers
and
percentages are specified to the nearest significant digit. Values of pH are
specified to
0.5.
[310] While exemplary pH ranges are given in some examples, other pH ranges
are
possible.
[311] The process may be scaled up using larger quantities and modified
apparatus.
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[312] The extraction process in other embodiments may use varying applied
pressures and temperatures, which vary during the soaking steps.
[313] Chemical and physical stability may be determined using rigorous
stability
testing protocols. This would be a necessary study for the product to be
considered
made using a good manufacturing process. The initial specifications and
ongoing
specifications of the extract should be determined during a testing regime
over time,
temperature, relative humidity, etc. to determine the physical and chemical
stability.
Studies are on-going, but a 5-year shelf life with a minimum of 2 years is
targeted in
terms of physical and chemical stability.
E. Purification
Purification Process 1
[314] In one embodiment, referring to FIG. 9, a basic process for obtaining
a
purified psychoactive alkaloid solution from extracted psychoactive compounds
is
shown. The process includes the step 900 of extracting a psychoactive alkaloid
from a
psychoactive alkaloid source to obtain a psychoactive alkaloid extract. The
psychoactive alkaloid source may be a fungus, a mycelium, an animal, a spore,
a plant,
a bacterium, or a yeast. The psychoactive alkaloid source in some embodiments
may
be a prior extract of one or more psychoactive alkaloids, where the prior
extract is to
undergo a further extraction/purification process. The psychoactive alkaloid
may
include, but is not limited to, psilocybin, psilocin, baeocystin,
norbaeocystin, norpsilocin,
aeruginascin, bufotenin, bufotenidine, 5-Me0-DMT (5-methoxy-N,N-
dimethyltryptamine), N,N-dimethyltryptamine (DMT), ergine (LSA), ergonovine,
ergometrine, muscimol, ibotenic acid, lysergic acid hydroxyethylamide (LSH),
elymoclavine, ergometrinine, and/or chanoclavine. or any combination
therefrom. The
extract from the psychoactive alkaloid source may be a fluid, as either a
liquid or a
slurry, or is made into a fluid by the addition of a solvent.
[315] The solvent in which the extract is carried or dissolved may be a
primary
aliphatic alcohol, a ketone, water, and any combination therefrom. In one
embodiment,
the primary aliphatic alcohol is a C1-4 alcohol. In one embodiment, the
primary
aliphatic alcohol is 5% ethanol. In one embodiment, the primary aliphatic
alcohol is
ethanol. In one embodiment, the ketone is a C3-4 ketone. In yet another
embodiment,
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the water is selected from deionized, distilled, reverse osmosis, or otherwise
purified
water, which is substantially without free ions. In other embodiments, the
water is not
purified.
[316] The process then involves adsorbing, in step 910, the psychoactive
alkaloid(s)
in the extract obtained in step 900 onto a resin to obtain an adsorbed
psychoactive
alkaloid, which may include one or more adsorbed psychoactive alkaloids.
[317] In one embodiment, the resin is an adsorbent resin of the macroporous
type,
such as, a cation or anion ion-exchange resin, a non-ionic resin, or any
combination
therefrom. Representative pharmaceutical, nutraceutical or food-grade grade
resins for
use in accordance with the present invention are known to those skilled in the
art. For
example, pharmaceutical grade non-ionic macroporous resins are commercially
available, e.g. Amberlite XAD4. In one embodiment, the resin is a cationic
ion-
exchange resin or an anionic-exchange resin. The cationic ion-exchange resin
may be
selected from commercially available cationic ion-exchange resins known in the
art,
including, but not limited to, Amberlite MAC-3 H. The cationic ion-exchange
resin may
be in an H+ form or an Na + form. The anionic ion-exchange resin may be
selected from
commercially available anion exchange resins known in the art, including, but
not
limited to, Amberchrom 50WX8. The anionic ion-exchange resin may be in an OH-
form or a Cl- form. The resins used may be of any particle size. In some
embodiments,
the resins may be gel type resins, with any size of gel bead.
[318] Next, the process involves eluting, in step 920, the adsorbed
psychoactive
alkaloid using a solvent to obtain a purified psychoactive alkaloid solution.
The solvent
may be, for example, an organic solvent, an acid, a base, a combination of an
organic
solvent and a base, a combination an organic solvent and an acid, water, a
combination
of water and acid, a combination of water and base, or a combination of water
and an
organic solvent. Usually, the solvent is different from the solvent in which
the extract is
initially provided and is either a different type of solvent or a different
composition of
solvent. It may be at a different temperature than the initial solvent.
[319] In some embodiments, the solvent used in the elution step 920 may be
a
primary aliphatic alcohol, a ketone, water, and any combination therefrom. In
one
embodiment, the primary aliphatic alcohol is a C1-4 alcohol. In one
embodiment, the
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primary aliphatic alcohol is 5% ethanol. In one embodiment, the primary
aliphatic
alcohol is ethanol. In one embodiment, the ketone is a C3-4 ketone. In yet
another
embodiment, the water is deionized, distilled, reverse osmosis, or otherwise
purified
water, which is substantially without free ions. In other embodiments, the
water is not
purified.
[320] In one embodiment, the solvent used in the elution step 920 is a
combination
of an organic solvent and an acid. In one embodiment, the combination of an
acid and
an organic solvent is 2% hydrochloric acid and 80% ethanol, for example. In
general,
any acidic environment will displace some of the ions from the resin, but the
rate and
optimization of the desorption will be affected by the acid concentration. In
one
embodiment, the solvent used in the elution step 920 is a combination of an
organic
solvent and a base. In one embodiment, the combination of an organic solvent
and a
base is of 2% sodium chloride and 80% ethanol, for example. In general, any
basic
environment will displace some of the ions from the resin, but the rate and
optimization
of the desorption will be affected by the concentration of the base.
[321] All the above solvents and combinations thereof are suitable for the
elution
step because all of the psychoactive alkaloids of interest are soluble therein
and,
depending on the choice of resin, they are all capable of pulling the
alkaloids of interest
off a resin. There are many different resins available, and it is a
straightforward matter
to select a suitable resin and elution solvent pair.
[322] In one embodiment, the elution step is carried out at a temperature
in the
range of 4-75 C. These temperatures are limited by the boiling point of the
solvent
used, as well as the specifications of allowable food-grade resins, as
determined by the
manufacturers of the resins and governmental food and drug administrations. In
another embodiment, the elution step is carried out at room temperature for
convenience, i.e., at 21-25 C.
[323] In other embodiments, the process for obtaining the purified
psychoactive
alkaloid solution further includes repeating the steps 910 and 920 with the
obtained
purified psychoactive alkaloid solution to obtain a further or twice purified
psychoactive
alkaloid solution. For the repeated steps in these embodiments, the resin may
be the
same or a different resin, and the solvent may be the same or a different
solvent. While
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the purified psychoactive alkaloid solution may have a low psychoactive
alkaloid
content, this may be increased by evaporation of some or all of the solvent.
Purification Process 2
[324] In some embodiments, referring to FIG. 16, a process for purifying
the
psychoactive alkaloid extract obtained in step 1410 (FIG. 14) or step 1520
(FIG. 15) is
shown.
[325] The process includes adding, in step 1600, an acid or a base to the
psychoactive alkaloid extract previously obtained, to result in a specific pH
psychoactive
alkaloid solution.
[326] When used, the acid may be acetic acid, adipic acid, ascorbic acid,
phosphoric acid, ammonium aluminum sulphate, ammonium citrate dibasic,
ammonium
citrate monobasic, calcium citrate, calcium fumarate, calcium gluconate,
calcium
phosphate dibasic, calcium phosphate monobasic, hydrochloric acid, sulfuric
acid
monobasic, calcium phosphate tribasic, citric acid, fumaric acid, gluconic
acid,
magnesium fumarate, malic acid, phosphoric acid, potassium acid tartrate,
potassium
citrate, potassium fumarate, sodium citrate, sodium fumarate, sodium
gluconate,
sodium lactate, sodium potassium hexametaphosphate, sodium potassium tartrate,
sodium potassium tripolyphosphate, sodium pyrophosphate tetrabasic, sodium
tripolyphosphate, tartaric acid, or any combination of one or more of these.
In some
embodiments, the acid is either only hydrochloric acid or only phosphoric
acid, for
example. It is also envisaged that other acids may be used.
[327] When used, the base may be ammonium bicarbonate, ammonium carbonate,
ammonium hydroxide, calcium acetate, calcium carbonate, calcium chloride,
calcium
hydroxide, calcium lactate, calcium oxide, calcium phosphate dibasic, calcium
phosphate monobasic, magnesium carbonate, potassium aluminum sulphate,
potassium bicarbonate, potassium carbonate, potassium hydroxide, potassium
lactate,
potassium phosphate dibasic, potassium pyrophosphate tetrabasic, potassium
phosphate tribasic, potassium tripolyphosphate, sodium acetate, sodium acid
pyrophosphate, sodium aluminum phosphate, sodium aluminum sulphate, sodium
bicarbonate, sodium bisulphate, sodium carbonate, sodium hexametaphosphate,
sodium hydroxide, sodium lactate, sodium phosphate dibasic, sodium phosphate
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monobasic, sodium phosphate tribasic, or any combination selected therefrom.
In one
embodiment, the base is solely sodium hydroxide, for example. Other bases may
be
used in other embodiments.
[328] In some embodiments, the specific pH psychoactive alkaloid solution
has a
pH ranging from 2.5 to 4.5 or from 9 to 10. In other embodiments, the specific
pH
psychoactive alkaloid solution has a pH of 3, 4, or 9.5. The selection of the
pH is
chosen in a manner to allow for the efficient adsorption of the psychoactive
alkaloids
onto the resin(s).
[329] The process optionally includes partially evaporating the solvent
from the
specific pH psychoactive alkaloid solution.
[330] The process optionally includes filtering, centrifuging, or
clarifying the
psychoactive alkaloid solution or specific pH psychoactive alkaloid solution,
as the case
may be, and utilizing the obtained filtrate for the next step 1610 of
adsorption.
Clarifying may be performed, for example, by adding an agglomeration agent.
[331] In step 1610, the process involves adsorbing the psychoactive
alkaloid(s) in
the specific pH psychoactive alkaloid solution onto a resin to obtain an
adsorbed
psychoactive alkaloid.
[332] In step 1620, the process involves washing the resin to remove
adsorbed
impurities from the resin. While not all the impurities are adsorbed onto the
resin, some
of them may be. The washing step, substantially, does not remove the adsorbed
psychoactive alkaloids. The washing solvent may be 100% ethanol, 100% reverse
osmosis water, or any other washing solvent used in ion-exchange resin
chromatography, provided that the washing removes impurities or by-products
that are
adsorbed on the resin. Impurities or by-products may include, for example,
sugars,
carbohydrates, chitin, chitosan, fats, minerals, waxes, or proteins. There may
be one,
two or more washing steps depending on the embodiment, and the same or
different
washing solvents may be used for each wash.
[333] In other embodiments, the solvent(s) for washing may include a
primary
aliphatic alcohol, a ketone, water, and any combination selected therefrom. In
some
embodiments, the primary aliphatic alcohol is a C1-4 alcohol. In some
embodiments,
the primary aliphatic alcohol is 5% ethanol. In some embodiments, the primary
aliphatic
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alcohol is ethanol. In some embodiments, the ketone is a C3-4 ketone. In yet
other
embodiments, the water is selected from deionized, distilled, reverse osmosis,
or
otherwise purified water that is substantially without free ions.
[334] After the washing, the process involves eluting, in step 1630, the
adsorbed
psychoactive alkaloid from the resin using a solvent to obtain a purified
psychoactive
alkaloid extract. The solvent may be an organic solvent, an acid, a base, or
water, a
combination of an organic solvent and a base, or a combination of an organic
solvent
and an acid, a combination of an organic solvent and water, a combination of
water and
a base, or combination of water and an acid. The solvent from the purified
psychoactive alkaloid extract is partially evaporated to obtain the purified
psychoactive
alkaloid extract in a slurry form for the standardization step 1420.
[335] In some embodiments, the solvent used in the elution step 1630 may be
a C1-
4 alcohol, a C3-4 ketone, water, and any combination selected therefrom. In
one
embodiment, the primary aliphatic alcohol is 5% ethanol. In yet another
embodiment,
the water is deionized, distilled, reverse osmosis, or otherwise purified
water, which is
substantially without free ions. In one embodiment, the solvent used in the
elution step
14 is a combination of an organic solvent and an acid. In general, any acidic
environment will displace some of the ions from the resin, but the rate and
optimization
of the desorption will be affected by the acid concentration. In one
embodiment, the
solvent used is a combination of an organic solvent and a base. In general,
any basic
environment will displace some of the ions from the resin, but the rate and
optimization
of the desorption will be affected by the concentration of the base.
[336] All the above solvents and combinations thereof are suitable for the
elution
step because all of the psychoactive alkaloids of interest are soluble therein
and,
depending on the choice of resin, they are all capable of pulling the
alkaloids of interest
off a resin. There are many different resins available, and it is a
straightforward matter
to select a suitable resin and elution solvent pair.
[337] In one embodiment, the elution step is carried out at a temperature
in the
range of 4-75 C. These temperatures are limited by the boiling point of the
solvent
used, as well as the specifications of allowable food-grade resins, as
determined by the
manufacturers of the resins and governmental food and drug administrations. In
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another embodiment, the elution step is carried out at room temperature for
convenience, i.e., at 21-25 C.
[338] In other embodiments, the process for obtaining the purified
psychoactive
alkaloid
[339] The extraction step further includes repeating the steps 1600 to 1630
with the
purified psychoactive alkaloid extract obtained in step 1630 to obtain a
further or twice
purified psychoactive alkaloid extract. For the repeated steps in these
embodiments,
the resin may be the same or a different resin, and the solvent may be the
same or a
different solvent. While the purified psychoactive alkaloid extract may have a
low
psychoactive alkaloid content, this may be increased by evaporation of some or
all of
the solvent.
F. Standardization
Standardization of Psychoactive Alkaloid 1
[340] In one embodiment, referring to FIG. 14, a process for obtaining the
composition is shown. The process includes the step 1400 of extracting a
psychoactive
alkaloid from a psychoactive alkaloid source to obtain a psychoactive alkaloid
extract.
The extract from the psychoactive alkaloid source may be a fluid, as either a
liquid or a
slurry, or is made into a fluid by the addition of a solvent. In one
embodiment, the
extraction step 1400 is followed by completely or partially concentrating the
obtained
psychoactive alkaloid extract (or solution) by evaporation of the solvent from
the extract.
In other embodiments, partially or completely evaporating the solvent may be
considered to be part of the extraction step 1400. If the solvent from the
extract has
been completely evaporated, then water, more solvent, or another solvent is
added
back.
[341] In step 1410, the extract obtained is filtered, followed by
concentration to
obtain a concentrated psychoactive alkaloid extract. Filtration is performed
by any
suitable known technique.
[342] In step 1420, the concentrated psychoactive alkaloid extract is then
standardized by adding one or more excipients to the extract, followed by
drying to
obtain the standardized psychoactive alkaloid composition. The concentration
of
alkaloids in the extract is measured, and the proportion of dry weight in the
extract is
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calculated. Based on this concentration and dry weight content, the amount of
excipient to be added is chosen to result in a powered composition with a
specific
amount of psychoactive alkaloid when the concentrated extract is dried.
[343] The drying can be achieved by any technique known in the art for
drying moist
compositions including, for example, spray drying or freeze drying.
Standardization of Psychoactive Alkaloid 2
[344] Referring to FIG. 11, the present invention also relates to a process
of
obtaining a standardized, purified, psychoactive alkaloid extract. In one
embodiment,
the process includes, in step 1100, concentrating the purified psychoactive
alkaloid
solution to obtain a purified psychoactive alkaloid slurry. The slurry is then
standardized, in step 1110, to obtain a standardized psychoactive alkaloid
extract.
[345] In one embodiment, the standardizing step 1110 includes adding
excipients to
the purified psychoactive alkaloid slurry to obtain the standardized
psychoactive
alkaloid extract. The concentration of alkaloids in the slurry is measured,
and the
proportion of dry weight in the slurry is calculated. Knowing this
concentration and the
dry weight content, the amounts of excipients are chosen to result in a powder
of known
alkaloid concentration after the solvent in the slurry has been evaporated.
[346] In one embodiment, the excipients are selected from silicon dioxide,
ascorbic
acid, maltodextrin from corn, potato or tapioca for example, gum arabic,
microcrystalline
cellulose, sodium benzoate, sodium phosphate, sodium citrate, rice hulls, and
rice. A
combination of any of these excipients may be used.
[347] Depending on the concentration of the purified psychoactive alkaloid
slurry
and the quantity of excipients added, the standardized psychoactive alkaloid
extract
may have a psychoactive alkaloid concentration ranging from 0.1-99% by weight,
and
the concentration may be specified to two decimal places or two significant
figures. For
the highest percentage, only 1% of the standardized psychoactive extract will
be
excipient.
[348] In exemplary embodiments, the standardized psychoactive alkaloid
extracts
have psychoactive alkaloid concentrations of 5.00% by weight, 54% by weight,
and
68% by weight.
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NUMBERED EMBODIMENTS
[349] 1. A psychoactive alkaloid composition comprising of, by weight: 0.1-
99.9%
of a psychoactive alkaloid extract; and one or more preservatives up to 10%, a
flow
agent up to 2%, 0-94% of one or more carriers, or any combination thereof.
[350] 2. The composition of embodiment 1, comprising 2-99.7% of the
psychoactive alkaloid extract.
[351] 3. The composition of embodiment 1, comprising an antioxidant up to
0.5%
by weight.
[352] 4. The composition of embodiment 1, comprising a bioavailability
agent up to
0.5% by weight.
[353] 5. The composition of embodiment 1, wherein the psychoactive alkaloid
extract has a psychoactive alkaloid concentration ranging from 0.1% to 99% by
weight
of the extract.
[354] 6. The composition of embodiment 5 wherein the one or more
preservatives
are selected from ascorbic acid, citric acid, lactose, vitamin A, vitamin E,
retinyl
palmitate, selenium, sodium citrate, sodium ascorbate, calcium ascorbate,
sodium
benzoate, and potassium benzoate.
[355] 7. The composition of embodiment 1, wherein the psychoactive alkaloid
extract has a psychoactive alkaloid concentration ranging from 0.1% to 20% by
weight
of the extract.
[356] 8. The composition of embodiment 1, wherein the psychoactive alkaloid
extract is a purified psychoactive alkaloid extract, and the purified
psychoactive alkaloid
extract has a psychoactive alkaloid concentration ranging from 10% to 99% by
weight.
[357] 9. The composition as in embodiment 1, in a powder form.
[358] 10. The composition of embodiment 1, comprising 10% or more of the
carrier.
[359] 11. The composition of embodiment 1, wherein the psychoactive
alkaloid is
psilocybin, psilocin, baeocystin, norbaeocystin, norpsilocin, aeruginascin,
bufotenin,
bufotenidine, 5-Me0-DMT (5-methoxy-N.Ndimethyltryptamine), N,N-
dimethyltryptamine
(DMT), 4-hydroxytryptamine, N,N,N-trimethy1-4-hydroxytryptamine ergine (LSA),
ergonovine, ergometrine, muscimol, ibotenic acid, lysergic acid
hydroxyethylamide
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(LSH), elymoclavine, ergometrinine, and/or chanoclavine, or any combination
selected
therefrom.
[360] 12. The composition of embodiment 1, wherein the psychoactive
alkaloid
extract comprises naturally occurring substances selected from the group
consisting of
fats, sugars, carbohydrates, chitin, chitosan, minerals, waxes and proteins.
[361] 13. The composition of embodiment 10, wherein the naturally occurring
substances are present in the psychoactive alkaloid extract in a concentration
ranging
from 1% - 99.9% by weight.
[362] 14. The composition of embodiment 1, wherein the psychoactive
alkaloid
extract is from fungi.
[363] 15. The composition of embodiment 14, wherein the psychoactive
alkaloid
extract is from Psilocybe cyanescens, Psilocybe cubensis, Amanita muscaria, or
any
selection therefrom.
[364] 16. The composition of embodiment 1, wherein the psychoactive
alkaloid
extract is from psychoactive plants.
[365] 17. The composition of embodiment 16, wherein the psychoactive
alkaloid
extract is from Anadenanthera colubrina.
[366] 18. The composition of embodiment 16, wherein the psychoactive
alkaloid
extract is from Anadenanthera peregrina.
[367] 19. The composition of embodiment 1, wherein the psychoactive
alkaloid
extract is from psychoactive animals.
[368] 20. The composition of embodiment 19, wherein the psychoactive
alkaloid
extract is from Incilius alvarius.
[369] 21. The composition of embodiment 1, wherein the psychoactive
alkaloid
extract is from psychoactive yeasts.
[370] 22. The composition of embodiment 1, wherein the flow agent is
selected
from silicon dioxide, stearic acid, magnesium stearate, or talc.
[371] 23. The composition of embodiment 1, wherein the one or more carriers
are
selected from starch, maltodextrin, alpha and beta cyclodextrin,
microcrystalline
cellulose (MCC), gum arabic, xanthum gum, guar gum, mannitol, or cellulose
gum.
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[372] 24. The composition of embodiment 23, wherein the maltodextrin is
tapioca
maltodextrin or rice maltodextrin.
[373] 25. The composition of embodiment 23, wherein the starch is potato
starch.
[374] 26. The composition of embodiment 1, wherein the flow agent is
present in
the composition at 0.1 to 1.2%.
[375] 27. The composition of embodiment 1, wherein a first preservative of
the one
or more preservatives is present in the composition at 0.1 to 3%.
[376] 28. The composition of embodiment 27, wherein a second preservative
of the
one or more preservatives is present in the composition at 0.1 to 3%.
[377] 29. The composition of embodiment 1, wherein a first carrier of the
one or
more carriers is present in the composition at 10 to 20%.
[378] 30. The composition of embodiment 29, wherein a second carrier of the
one
or more carriers is present in the composition at 10 to 20%.
[379] 31. The composition of embodiment 1, comprising: the flow agent
present in
the composition at 0.1 to 1.2%; the one or more preservatives present in the
composition at 0.1 to 2%; and the one or more carriers present in the
composition at 10
to 20%.
[380] 32. The composition of embodiment 1, wherein: the flow agent is
silicon
dioxide; the carrier comprises maltodextrin and mannitol; and the one or more
preservatives comprise ascorbic acid and citric acid.
[381] 33. The composition of embodiment 32, wherein the silicon dioxide is
present
in the composition at 0.1 to 1.2%.
[382] 34. The composition of embodiment 32, wherein the ascorbic acid is
present
in the composition at 0.1 to 2%.
[383] 35. The composition of embodiment 32, wherein the citric acid is
present in
the composition at 0.1 to 2%.
[384] 36. The composition of embodiment 32, wherein the maltodextrin is
present in
the composition at 10 to 20%.
[385] 37. The composition of embodiment 32, wherein the mannitol is present
in the
composition at 10 to 20%.
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[386] 38. The composition of embodiment 1, wherein: the flow agent is
silicon
dioxide; the carrier comprises starch and mannitol; and the one or more
preservatives
comprise ascorbic acid and citric acid.
[387] 39. The composition of embodiment 38, wherein the silicon dioxide is
present
in the composition at 0.1 to 1.2%.
[388] 40. The composition of embodiment 38, wherein the ascorbic acid is
present
in the composition at 0.1 to 2%.
[389] 41. The composition of embodiment 38, wherein the citric acid is
present in
the composition at 0.1 to 2%.
[390] 42. The composition of embodiment 38, wherein the starch is present
in the
composition at 10 to 20%.
[391] 43. The composition of embodiment 38, wherein the mannitol is present
in the
composition at 10 to 20%.
[392] 44. A method for generating a psychoactive alkaloid extract from a
psychoactive organism, the method comprising: a. providing a biomass of the
psychoactive organism; b. contacting the biomass with 10 to 100 milliliters
(mL) of
solvent per gram (g) of the biomass; and c. evaporating the solvent from the
biomass to
yield the psychoactive alkaloid extract.
[393] 45. The method of embodiment 44, wherein the solvent is selected from
100% methanol, an alcohol:water mixture wherein the alcohol comprises 60% to
99% of
the alcohol:water mixture, an alcohol:acid mixture wherein the alcohol
comprises 60%
to 99% of the alcohol:acid mixture, and acidified water.
[394] 46. A method for generating a psychoactive alkaloid extract from a
psychoactive organism, the method comprising: a. providing a biomass of the
psychoactive organism; b. contacting the biomass with a solvent, wherein the
solvent is
selected from 100% methanol, an alcohol:water mixture wherein the alcohol
comprises
60% to 99% of the alcohol:water mixture, an alcohol:acid mixture wherein the
alcohol
comprises 60% to 99% of the alcohol:acid mixture, and acidified water; and c.
evaporating the solvent from the biomass to yield the psychoactive alkaloid
extract.
[395] 47. The method of embodiment 46, wherein the solvent is present at 10
to
100 milliliters (mL) of solvent per gram (g) of the biomass.
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[396] 48. The method of any one of embodiments 44 to 47, wherein the
solvent is
present at 10 to 60 mL of solvent per gram of the biomass.
[397] 49. The method of any one of embodiments 44 to 48, wherein the
solvent is
present at 40 to 60 mL of solvent per gram of the biomass.
[398] 50. The method of any one of embodiments 44 to 49, wherein the
biomass of
the psychoactive organism is dried prior to contacting with the solvent.
[399] 51. The method of any one of embodiments 44 to 50, wherein the
biomass of
the psychoactive organism is reduced to a particle size of 6 millimeters (mm)
to 0.03
mm prior to contacting with the solvent.
[400] 52. The method of any one of embodiments 44 to 51, wherein the
biomass of
the psychoactive organism is reduced to a particle size of 1 mm to 0.03 mm
prior to
contacting with the solvent.
[401] 53. The method of any one of embodiments 44 to 52, wherein the
biomass of
the psychoactive organism is reduced to a particle size of at least 0.074 mm
prior to
contacting with the solvent.
[402] 54. The method of any one of embodiments 44 to 53, wherein the
biomass of
the psychoactive organism is contacted with the solvent at 5 C to 95 C.
[403] 55. The method of any one of embodiments 44 to 54, wherein the
biomass of
the psychoactive organism is contacted with the solvent at 20 C to 70 C.
[404] 56. The method of any one of embodiments 44 to 55, wherein the
biomass of
the psychoactive organism is contacted with the solvent at 25 C.
[405] 57. The method of any one of embodiments 44 to 56, wherein the
biomass of
the psychoactive organism is contacted with the solvent for 1 to 720 minutes.
[406] 58. The method of any one of embodiments 44 to 57, wherein the
biomass of
the psychoactive organism is contacted with the solvent for 20 to 60 minutes.
[407] 59. The method of any one of embodiments 44 to 58, wherein the
biomass of
the psychoactive organism is contacted with the solvent for 30 minutes.
[408] 60. The method of any one of embodiments 44 to 59, wherein, following
(b),
the biomass is filtered through a filter.
[409] 61. The method of embodiment 60, wherein the filter comprises 1
micron (pm)
to 10 pm mesh.
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[410] 62. The method of any one of embodiments 44 to 61, wherein, following
(b),
the biomass is contracted with a second solvent.
[411] 63. The method of embodiment 62, wherein the second solvent is
selected
from 100% methanol, an alcohol:water mixture wherein the alcohol comprises 60%
to
99% of the alcohol:water mixture, an alcohol:acid mixture wherein the alcohol
comprises 60% to 99% of the alcohol:acid mixture, and acidified water.
[412] 64. The method of embodiment 62 or 63, wherein the second solvent is
present at 10 to 100 milliliters (mL) of solvent per gram (g) of the biomass.
[413] 65. The method of any one of embodiments 62 to 64, wherein the
solvent is
present at 10 to 60 mL of solvent per gram of the biomass.
[414] 66. The method of any one of embodiments 62 to 65, wherein the
solvent is
present at 40 to 60 mL of solvent per gram of the biomass.
[415] 67. The method of any one of embodiments 44 to 66, wherein the
alcohol of
the alcohol:water mixture, the alcohol:acid mixture, or both, is a C1-C4
primary aliphatic
alcohol.
[416] 68. The method of embodiment 67, wherein the C1-C4 primary aliphatic
alcohol is ethanol or methanol.
[417] 69. The method of any one of embodiments 44 to 68, wherein the acid
in the
alcohol:acid mixture, the acidified water, or both, is acetic acid, adipic
acid, ascorbic
acid, phosphoric acid, ammonium aluminum sulphate, ammonium citrate dibasic,
ammonium citrate monobasic, calcium citrate, calcium fumarate, calcium
gluconate,
calcium phosphate dibasic, calcium phosphate, hydrochloric acid, sulphuric
acid
monobasic, calcium phosphate tribasic, citric acid, fumaric acid, gluconic
acid,
magnesium fumarate, malic acid, phosphoric acid, potassium acid tartrate,
potassium
citrate, potassium fumarate, sodium citrate, sodium fumarate, sodium
gluconate,
sodium lactate, sodium potassium hexametaphosphate, sodium potassium tartrate,
sodium potassium tripolyphosphate, sodium pyrophosphate tetrabasic, sodium
tripolyphosphate, tartaric acid, and any combination therefrom.
[418] 70. The method of any one of embodiments 44 to 68, wherein the
solvent,
second solvent, or both is buffered to a pH of either 4 or less, or 10 or
greater.
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[419] 71. The method of embodiment 70, wherein the solvent, second solvent,
or
both is buffered with ammonium bicarbonate, ammonium carbonate, ammonium
hydroxide, calcium acetate, calcium carbonate, calcium chloride, calcium
hydroxide,
calcium lactate, calcium oxide, calcium phosphate, dibasic, calcium phosphate
monobasic, magnesium carbonate, potassium aluminum sulphate, potassium
bicarbonate, potassium carbonate, potassium hydroxide, potassium lactate,
potassium
phosphate, dibasic, potassium pyrophosphate, tetrabasic, potassium phosphate
tribasic, potassium tripolyphosphate, sodium acetate, sodium acid
pyrophosphate,
sodium aluminum phosphate, sodium aluminum sulphate, sodium bicarbonate,
sodium
bisulphate, sodium carbonate, sodium hexametaphosphate, sodium hydroxide,
sodium
lactate, sodium phosphate dibasic, sodium phosphate monobasic, sodium
phosphate
tribasic, and any combination therefrom.
[420] 72. The method of embodiment 70 or 71, wherein the solvent, second
solvent,
or both is buffered with acetic acid, adipic acid, ascorbic acid, phosphoric
acid,
ammonium aluminum sulphate, ammonium citrate dibasic, ammonium citrate
monobasic, calcium citrate, calcium fumarate, calcium gluconate, calcium
phosphate
dibasic, calcium phosphate, hydrochloric acid, sulphuric acid monobasic,
calcium
phosphate tribasic, citric acid, fumaric acid, gluconic acid, magnesium
fumarate, malic
acid, phosphoric acid, potassium acid tartrate, potassium citrate, potassium
fumarate,
sodium citrate, sodium fumarate, sodium gluconate, sodium lactate, sodium
potassium
hexametaphosphate, sodium potassium tartrate, sodium potassium
tripolyphosphate,
sodium pyrophosphate tetrabasic, sodium tripolyphosphate, tartaric acid, and
any
combination therefrom.
[421] 73. The method of any one or embodiments 44 to 72, wherein the
psychoactive alkaloid is psilocybin, psilocin, baeocystin, norbaeocystin,
norpsilocin,
aeruginascin, bufotenin, bufotenidine, 5-Me0-DMT (5-methoxy-
N.Ndimethyltryptamine),
N,N-dimethyltryptamine (DMT), 4-hydroxytryptamine, N,N,N-trimethy1-4-
hydroxytryptamine ergine (LSA), ergonovine, ergometrine, muscimol, ibotenic
acid,
lysergic acid hydroxyethylamide (LSH), elymoclavine, ergometrinine, and/or
chanoclavine, or any combination selected therefrom.
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[422] 74. The method of any one or embodiments 44 to 72, wherein the
solvent
comprises a pH of 10 or greater and the psychoactive alkaloid extract
comprises
greater than 50% of the phosphorylated psychoactive alkaloid.
[423] 75. The method of embodiment 74, wherein the psychoactive alkaloid
extract
comprises greater than 90% of a phosphorylated psychoactive alkaloid.
[424] 76. The method of embodiment 74 or 75, wherein the phosphorylated
alkaloid
is psilocybin, baeocystin, norbaeocystin, aeruginascin, or any combination
therefrom.
[425] 77. The method of any one or embodiments 44 to 72, wherein the
solvent
comprises a pH of 4 or less and the psychoactive alkaloid extract comprises
greater
than 50% of a dephosphorylated psychoactive alkaloid.
[426] 78. The method of embodiment 77, wherein the psychoactive alkaloid
extract
comprises greater than 90% of the dephosphorylated psychoactive alkaloid.
[427] 79. The method of embodiment 77 or 78, wherein the dephosphorylated
alkaloid is psilocin, norpsilocin, 4-hydroxytryptamine, N,N,N-trimethy1-4-
hydroxytryptamine, or any combination therefrom.
[428] 80. The method of any one or embodiments 44 to 79, wherein the
psychoactive organism is a plant, animal, fungus, Protist, or bacterium.
[429] 81. The method of any one or embodiments 44 to 80, wherein the
psychoactive organism is Psilocybe cyanescens, Psilocybe cubensis, Amanita
muscaria, or any selection therefrom.
[430] 82. The method of any one or embodiments 44 to 80, wherein the
psychoactive organism is Anadenanthera colubrina or Anadenanthera peregrina.
[431] 83. The method of any one or embodiments 44 to 80, wherein the
psychoactive organism is Incilius alvarius.
[432] 84. The method of any one or embodiments 44 to 80, wherein the
psychoactive organism is yeast.
[433] 85. A process for forming an extract of psychoactive alkaloids from
psychoactive organisms comprising the steps of: soaking a biomass of dried,
raw
psychedelic fungus in a solvent selected from the group consisting of ethanol,
a water-
ethanol mixture, methanol, and a water-methanol mixture in order to dissolve
the
psychoactive alkaloids in the solvent; filtering an undissolved portion of the
biomass
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from the solvent; evaporating the solvent sufficiently to remove the solvent
completely,
leaving a concentrated slurry or a residue that is converted to the
concentrated slurry by
adding water thereto: and standardizing the concentrated slurry by adding
thereto a
quantity of carrier measured to achieve a specified purity of extract.
[434] 86. The process of embodiment 85, wherein the standardizing
comprises:
measuring a psychoactive alkaloid content in the concentrated slurry; and
using the
psychoactive alkaloid content, the specified purity and a volume of the
concentrated
slurry to determine the quantity of carrier.
[435] 87. The process of embodiment 85, comprising drying the concentrated
slurry
to result in the extract, wherein the extract is a powdered extract.
[436] 88. The process of embodiment 85, wherein the solvent is a water-
ethanol or
water-methanol alkaline buffered solution.
[437] 89. The process of embodiment 88, wherein the solvent has a pH of 11-
12.
[438] 90. The process of embodiment 88, wherein the solvent is buffered
with
sodium hydroxide, the process comprising, between the filtering and
evaporating steps,
adjusting the solvent to a pH of 4-9 using phosphoric acid.
[439] 91. The process of embodiment 85, wherein the solvent is a water-
ethanol or
water-methanol acid buffered solution.
[440] 92. The process of embodiment 91, wherein the solvent has a pH of 1.8-
3.
[441] 93. The process of embodiment 91, wherein the solvent is buffered
with citric
acid, the process comprising, between the filtering and evaporating steps,
adjusting the
solvent to a pH of 4-9 using sodium hydroxide.
[442] 94. The process of embodiment 85, wherein the solvent comprises 100%
reverse osmosis water.
[443] 95. The process of embodiment 85, wherein the soaking is at a
temperature
of 5-95 C.
[444] 96. The process of embodiment 85, comprising applying a pressure of
50 kPa
¨ 100 MPa to the solvent during the soaking step.
[445] 97. The process of embodiment 85, comprising agitating the solvent
during
the soaking step, wherein the soaking step has a duration of 10 minutes to 12
hours.
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[446] 98. The process of embodiment 85, wherein the psychedelic organism is
a
plant, animal, fungus, Protist, or bacterium.
[447] 99. The process of embodiment 93, wherein the fungus comprises
amanita
muscaria, psilocybe cubensis, psilocybe cyanescens, or any combination
thereof.
[448] 100. The process of embodiment 85, wherein the psychoactive alkaloids
comprise psilocybin, psilocin, baeocystin, norbaeocystin, ibotenic acid or any
mixture
thereof.
[449] 101. The process of embodiment 85, wherein the solvent to biomass
ratio is in
a range from 11_:1kg to 50L1kg.
[450] 102. The process of embodiment 85, wherein the specified purity is
0.1-10%.
[451] 103. The process of embodiment 85, wherein the specified purity is
specified
as a percentage with a precision of two decimal places.
[452] 104. The process of embodiment 85, wherein the carrier comprises
ascorbic
acid, silicon dioxide, maltodextrin, gum arabic, microcrystalline cellulose,
sodium citrate,
sodium benzoate, sodium phosphate, rice, rice hulls, or any combination of the
foregoing.
[453] 105. The process of embodiment 85, comprising: repeating, using
further
solvent, the soaking and filtering steps for the filtered biomass; and
combining the
filtered solvent with the filtered further solvent.
[454] 106.A process for obtaining a purified psychoactive alkaloid
solution, the
process comprising: extracting a psychoactive alkaloid from a psychoactive
alkaloid
source to obtain a psychoactive alkaloid extract; contacting the psychoactive
alkaloid
extract with an adsorbent material to obtain an adsorbed psychoactive
alkaloid; and
eluting the adsorbed psychoactive alkaloid using a solvent to obtain a
purified
psychoactive alkaloid solution, wherein the solvent is water, an organic
solvent or a
combination thereof, under basic, acidic or neutral pH.
[455] 107. The process of embodiment 106, comprising, prior to the treating
step,
adding an acid or a base to the psychoactive alkaloid extract.
[456] 108. The process of embodiment 107, wherein, after adding the acid or
base,
the psychoactive alkaloid extract has a pH ranging from 2.5-4.5 or from 9-10
respectively.
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[457] 109. The process of embodiment 107, wherein the acid is selected from
the
group consisting of acetic acid, adipic acid, ascorbic acid, phosphoric acid,
ammonium
aluminum sulphate, ammonium citrate dibasic, ammonium citrate monobasic,
calcium
citrate, calcium fumarate, calcium gluconate, calcium phosphate dibasic,
calcium
phosphate, hydrochloric acid, sulphuric acid monobasic, calcium phosphate
tribasic,
citric acid, fumaric acid, gluconic acid, magnesium fumarate, malic acid,
phosphoric
acid, potassium acid tartrate, potassium citrate, potassium fumarate, sodium
citrate,
sodium fumarate, sodium gluconate, sodium lactate, sodium potassium
hexametaphosphate, sodium potassium tartrate, sodium potassium
tripolyphosphate,
sodium pyrophosphate tetrabasic, sodium tripolyphosphate, tartaric acid, and
any
combination therefrom.
[458] 110. The process of embodiment 107, wherein the base is selected from
the
group consisting of ammonium bicarbonate, ammonium carbonate, ammonium
hydroxide, calcium acetate, calcium carbonate, calcium chloride, calcium
hydroxide,
calcium lactate, calcium oxide, calcium phosphate, dibasic, calcium phosphate
monobasic, magnesium carbonate, potassium aluminum sulphate, potassium
bicarbonate, potassium carbonate, potassium hydroxide, potassium lactate,
potassium
phosphate, dibasic, potassium pyrophosphate, tetrabasic, potassium phosphate
tribasic, potassium tripolyphosphate, sodium acetate, sodium acid
pyrophosphate,
sodium aluminum phosphate, sodium aluminum sulphate, sodium bicarbonate,
sodium
bisulphate, sodium carbonate, sodium hexametaphosphate, sodium hydroxide,
sodium
lactate, sodium phosphate dibasic, sodium phosphate monobasic, sodium
phosphate
tribasic, and any combination therefrom.
[459] 111. The process of embodiment 106, wherein the adsorbent material is
a gel
resin, a macroporous resin, or a combination thereof.
[460] 112. The process of embodiment 111, wherein the macroporous resin is
a
non-ionic macroporous resin, an ion-exchange macroporous resin, or a
combination
thereof.
[461] 113. The process of embodiment 106, wherein the psychoactive alkaloid
source comprises psilocybin, psilocin, baeocystin, norbaeocystin, norpsilocin,
aeruginascin, bufotenin, bufotenidine, 5-Me0-DMT (5-methoxy-N.N-
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dimethyltryptamine), N,N-dimethyltryptamine (DMT), ergine (LSA), ergonovine,
ergometrine, muscimol, ibotenic acid, lysergic acid hydroxyethylamide (LSH),
elymoclavine, ergometrinine, chanoclavine, or any combination therefrom.
[462] 114. The process of embodiment 106, wherein the organic solvent is
selected
from a group consisting of C1-4 primary aliphatic alcohols, C3-4 ketones, and
any
combination therefrom.
[463] 115. The process of embodiment 106, wherein the process comprises
further
purifying the obtained purified psychoactive alkaloid solution by repeating,
with the
obtained purified psychoactive alkaloid solution, the treating step with a
different
adsorbent material and the eluting step with another solvent.
[464] 116. The process of embodiment 106, comprising: evaporating a portion
of
solvent from the purified psychoactive alkaloid solution to obtain a purified
psychoactive
alkaloid slurry.
[465] 117. The process of embodiment 116, wherein the purified psychoactive
alkaloid slurry comprises 5% by weight or more of a psychoactive alkaloid.
[466] 118. The process of embodiment 116, comprising:
[467] standardizing the purified psychoactive alkaloid slurry by adding
thereto a
quantity of excipient measured to provide a specific concentration of
psychoactive
alkaloid when the purified psychoactive alkaloid slurry is dried; and drying
the purified
psychoactive alkaloid slurry by evaporating the remaining portion of the
solvent to
obtain a standardized, purified, powdered psychoactive alkaloid extract having
the
specific concentration of psychoactive alkaloid.
[468] 119. The process of embodiment 116, wherein: the psychoactive
alkaloid
comprises psilocybin, psilocin, baeocystin, norbaeocystin, norpsilocin,
aeruginascin,
bufotenin, bufotenidine, 5-Me0-DMT (5-methoxy-N.N-dimethyltryptamine), N,N-
dimethyltryptamine (DMT), ergine (LSA), ergonovine, ergometrine, muscimol,
ibotenic
acid, lysergic acid hydroxyethylamide (LSH), elymoclavine, ergometrinine,
chanoclavine, or any combination therefrom; and the standardized, purified,
powdered
psychoactive alkaloid extract has a psychoactive alkaloid concentration
ranging from
0.1-99% by weight.
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[469] 120. The process of embodiment 118, wherein the excipient is selected
from
the group consisting of silicon dioxide, ascorbic acid, maltodextrin, gum
arabic,
microcrystalline cellulose, sodium benzoate, sodium phosphate, sodium citrate,
rice
hulls, rice and any combination therefrom.
[470] 121. The process of embodiment 106, comprising: prior to the treating
step:
adding an acid to the psychoactive alkaloid extract to bring its pH to 4 0.5;
and
removing solids from the psychoactive alkaloid extract; and after the treating
step and
before the eluting step: washing the adsorbent material with purified water;
wherein:
the adsorbent material is a non-ionic macroporous resin; and the solvent used
for the
eluting step is a hydro-ethanol solvent.
[471] 122. The process of embodiment 121, wherein the hydro-ethanol solvent
is
5% ethanol.
[472] 123. The process of embodiment 106, comprising: prior to the treating
step:
adding an acid to the psychoactive alkaloid extract to bring its pH to 3 0.5;
after the
treating step and before the eluting step: washing the adsorbent material with
100%
ethanol, wherein the adsorbent material is a macroporous strong cation
exchange resin
in an H+ or an Na+ form; and washing the adsorbent material with purified
water;
wherein the solvent used for the eluting step is 2% hydrochloric acid and 80%
ethanol;
and after the eluting step: adding alkali to the purified psychoactive
alkaloid solution to
bring its pH to 4 0.5; removing solids from the purified psychoactive alkaloid
solution;
evaporating a portion of the solvent from the purified psychoactive alkaloid
solution;
removing further solids from the purified psychoactive alkaloid solution;
treating the
purified psychoactive alkaloid extract with a non-ionic macroporous resin to
obtain a
second adsorbed psychoactive alkaloid; washing the non-ionic macroporous resin
with
purified water; and eluting the second adsorbed psychoactive alkaloid from the
non-
ionic macroporous resin using a hydro-ethanol solvent to obtain a twice
purified
psychoactive alkaloid solution.
[473] 124. The process of embodiment 106, comprising: prior to the treating
step:
adding a base to the psychoactive alkaloid extract to bring its pH to 9.5 0.5;
after the
treating step and before the eluting step: washing the adsorbent material with
100%
ethanol, wherein the adsorbent material is a macroporous strong anion exchange
resin
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in an OH- or a Cl- form; and washing the adsorbent material with purified
water;
wherein the solvent used for the eluting step is 2% sodium chloride and 80%
ethanol;
and after the eluting step: adding acid to the purified psychoactive alkaloid
solution to
bring its pH to 4 0.5; removing solids from the purified psychoactive alkaloid
solution;
evaporating a portion of the solvent from the purified psychoactive alkaloid
solution;
removing further solids from the purified psychoactive alkaloid solution;
treating the
purified psychoactive alkaloid extract with a non-ionic macroporous resin to
obtain a
second adsorbed psychoactive alkaloid; washing the non-ionic macroporous resin
with
purified water; and eluting the second adsorbed psychoactive alkaloid from the
non-
ionic macroporous resin using a hydro-ethanol solvent to obtain a twice
purified
psychoactive alkaloid solution.
[474] 125. The process of embodiment 106, wherein the psychoactive alkaloid
source comprises psychoactive fungus and the extracting step comprises: drying
and
pulverizing the psychoactive alkaloid source to obtain a dried biomass;
heating the
dried biomass in a first solvent to obtain a first slurry, and filtering the
first slurry to
obtain a first filtrate and a first residue; heating the first residue in a
second solvent to
obtain a second slurry, and filtering the second slurry to obtain a second
filtrate and a
second residue; and mixing the first filtrate and the second filtrate to
obtain the
psychoactive alkaloid extract.
[475] 126. The process of embodiment 125, wherein: the first solvent and
the
second solvent are selected from a group consisting of a primary aliphatic
alcohol, a
ketone, purified water, and any combination therefrom; and the heating is
carried out at
a temperature ranging from 5-95 C and for a time duration ranging from 10
minutes to
12 hours.
[476] 127. The process of embodiment 106, wherein the psychoactive alkaloid
source is Anadenanthera peregrina, the process comprising: prior to the
treating step:
adding an acid to the psychoactive alkaloid extract to bring its pH to 4 0.5;
and
removing solids from the psychoactive alkaloid extract; and after the treating
step and
before the eluting step: washing the adsorbent material with purified water
then with
10% ethanol; wherein: the adsorbent material is a macroporous resin; and the
solvent
used for the eluting step is 50% ethanol.
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[477] 128. The process of embodiment 106, wherein the psychoactive alkaloid
source comprises a plant, animal, fungus, protist, or bacterium.
[478] 129.T The process of embodiment 106, wherein the psychoactive
alkaloid
source comprises Psilocybe cyanescens, Psilocybe cubensis, Amanita muscaria,
or
any selection therefrom.
[479] 130. The process of embodiment 106, wherein the psychoactive alkaloid
source comprises Anadenanthera colubrina or Anadenanthera peregrina.
[480] 131. The process of embodiment 106, wherein the psychoactive alkaloid
source comprises Incilius alvarius.
[481] 132. The process of embodiment 106, wherein the psychoactive alkaloid
source comprises yeast.
[482] 133. The process of embodiment 106, wherein the psychoactive alkaloid
extract is contacted with the adsorbent material at a flow rate of 1 bed
volume per hour
(BV/h) to 10 BV/h.
[483] 134. The process of embodiment 133, wherein the psychoactive alkaloid
extract is contacted with the adsorbent material at a flow rate of 2 bed
volume per hour
(BV/h) to 6 BV/h.
[484] 135.A process for obtaining a psychoactive alkaloid extract with a
desired
amount of a phosphorylated psychoactive alkaloid and a desired amount of a
dephosphorylated psychoactive alkaloid, the process comprising: drying and
pulverizing a psychoactive alkaloid source to obtain a dried powdered biomass;
extracting a psychoactive alkaloid from the dried powdered biomass with an
acidified
solvent or a basified solvent to obtain a psychoactive alkaloid liquid with a
specific pH,
wherein the specific pH is lower than 3.5 or greater than 10.5; adjusting the
pH of the
psychoactive alkaloid liquid to a pH ranging from 3.5-4.5; and evaporating the
solvent
from the psychoactive alkaloid liquid to obtain the psychoactive alkaloid
extract with the
desired amount of the phosphorylated psychoactive alkaloid and the desired
amount of
the dephosphorylated psychoactive alkaloid; wherein: the desired amount of the
phosphorylated psychoactive alkaloid is 0-100% by weight of a total
phosphorylatable
psychoactive alkaloid content in the psychoactive alkaloid extract; and the
desired
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amount of the dephosphorylated psychoactive alkaloid is the remainder of the
total
phosphorylatable psychoactive alkaloid content in the psychoactive alkaloid
extract.
[485] 136. The process of embodiment 135, wherein the phosphorylated
alkaloid is
psilocybin, baeocystin, norbaeocystin, aeruginascin, or any combination
therefrom; and
the dephosphorylated alkaloid is psilocin, norpsilocin, 4-hydroxytryptamine,
N,N,N-
trimethy1-4-hydroxytryptamine, or any combination therefrom.
[486] 137. The process of embodiment 135, wherein the psychoactive alkaloid
source comprises psilocybin, baeocystin, norbaeocystin, aeruginascin,
psilocin,
norpsilocin, 4-hydroxytryptamine, N,N,N-trimethy1-4-hydroxytryptamine, or any
combination therefrom.
[487] 138. The process of embodiment 135, wherein the extracting step
comprises:
mixing the dried powdered biomass with the acidified solvent or the basified
solvent to
obtain a slurry; and filtrating the slurry to obtain a filtrate residue and
the psychoactive
alkaloid liquid.
[488] 139. The process of embodiment 138, wherein the extracting step
comprises
further extracting the psychoactive alkaloid by repeating, with the obtained
filtrate
residue, the extracting step with the same or a different acidified solvent,
or the same or
a different basified solvent.
[489] 140. The process of embodiment 138, wherein after the mixing step the
acidified solvent or the basified solvent, the slurry has a pH ranging from
0.5-3.5 or from
10.5-13.5 respectively.
[490] 141. The process of embodiment 135, wherein the acidified solvent is
a
mixture of an acid; and a C1-C4 primary aliphatic alcohol, a C3-C4 ketone,
water, or
any combination selected therefrom.
[491] 142. The process of embodiment 135, wherein the basified solvent is a
mixture of a base; and a C1-C4 primary aliphatic alcohol, a C3-C4 ketone,
water, or any
combination selected therefrom.
[492] 143. The process of embodiment 135, wherein the extraction is
performed: at
a temperature ranging from 5-95 C; and for a time period ranging from 10-720
minutes.
[493] 144. The process of embodiment 135, wherein the extraction is
performed at a
pressure ranging from 7 to 20,000 psi (50kPa ¨ 138MPa).
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[494] 145. The process of embodiment 135, wherein the extraction is
performed
with a solvent to solid proportion of 1L:lkg to 50L:lkg, wherein the solid is
the dried
powdered biomass.
[495] 146. The process of embodiment 135, wherein the specific pH is lower
than
3.5; and wherein: the desired amount of the phosphorylated psychoactive
alkaloid is
0% by weight of the total phosphorylatable psychoactive alkaloid content in
the
psychoactive alkaloid extract, and the desired amount of the dephosphorylated
psychoactive alkaloid is 100% by weight of the total phosphorylatable
psychoactive
alkaloid content in the psychoactive alkaloid extract.
[496] 147. The process of embodiment 135, wherein the specific pH is
greater than
10.5; and wherein the desired amount of the phosphorylated psychoactive
alkaloid is
100% by weight of the total phosphorylatable psychoactive alkaloid content in
the
psychoactive alkaloid extract, and the desired amount of the dephosphorylated
psychoactive alkaloid is 0% by weight of the total phosphorylatable
psychoactive
alkaloid content in the psychoactive alkaloid extract.
[497] 148. The process of embodiment 135, wherein the specific pH is
greater than
10.5, and a maximum desired amount of the phosphorylated alkaloid is limited
by an
amount of the dephosphorylated alkaloid in the psychoactive alkaloid source.
[498] 149. The process of embodiment 135, wherein the specific pH is
greater than
10.5, and wherein: the desired amount of the phosphorylated psychoactive
alkaloid is
1-99% by weight of the total phosphorylatable psychoactive alkaloid content in
the
psychoactive alkaloid extract.
[499] 150. The process of embodiment 135, comprising: pausing the
evaporating
step when a portion of the solvent has been evaporated from the psychoactive
alkaloid
liquid to obtain a psychoactive alkaloid slurry; standardizing the
psychoactive alkaloid
slurry by adding thereto a measured quantity of one or more excipients to
obtain a
standardized slurry with a specific amount of psychoactive alkaloid content;
and
continuing the evaporating step by drying the standardized slurry to obtain a
psychoactive alkaloid composition comprising the psychoactive alkaloid
extract, and
one or more excipients; wherein a total psychoactive alkaloid content in the
psychoactive alkaloid composition is specified as a result of the
standardizing step.
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[500] 151. The process of embodiment 150, wherein the desired amount of the
phosphorylated psychoactive alkaloid is 100% by weight of the total
phosphorylatable
psychoactive alkaloid content in the psychoactive alkaloid composition, the
process
comprising: preparing another psychoactive alkaloid composition comprising
another
psychoactive alkaloid extract according to embodiment 150, wherein the desired
amount of the dephosphorylated psychoactive alkaloid is 100% by weight of the
total
phosphorylatable psychoactive alkaloid content in the other psychoactive
alkaloid
extract; mixing the psychoactive alkaloid composition and the other
psychoactive
composition in a measured ratio to obtain a psychoactive alkaloid composition
comprising the phosphorylated psychoactive alkaloid of the psychoactive
alkaloid
composition and the dephosphorylated psychoactive alkaloid of the other
psychoactive
alkaloid composition in a specific ratio; wherein the specific ratio of
phosphorylated
psychoactive alkaloid to dephosphorylated psychoactive alkaloid ranges from
1:1000 to
1000:1.
[501] 152.A process for obtaining a psychoactive alkaloid composition with
a
specific ratio of a phosphorylated psychoactive alkaloid to a dephosphorylated
psychoactive alkaloid, the process comprising: extracting a psychoactive
alkaloid from
a dried powdered biomass with a basified solvent to obtain a psychoactive
alkaloid
liquid with a pH greater than 10.5, wherein a majority of a total
phosphorylatable
psychoactive alkaloid content is the phosphorylated alkaloid and a remainder
thereof is
the dephosphorylated alkaloid; adjusting the pH of the psychoactive alkaloid
liquid to a
pH ranging from 3.5-4.5; extracting another psychoactive alkaloid from another
dried
powdered biomass with an acidified solvent to obtain another psychoactive
alkaloid
liquid with a pH lower than 3.5, wherein all of a total phosphorylatable
psychoactive
alkaloid is the dephosphorylated alkaloid; adjusting the pH of the other
psychoactive
alkaloid liquid to a pH ranging from 3.5-4.5; evaporating a portion of the
basified
solvent from the psychoactive alkaloid liquid and a portion of the acidified
solvent from
the other psychoactive alkaloid liquid to obtain a psychoactive alkaloid
extract slurry
and another psychoactive alkaloid extract slurry respectively; mixing measured
portions of the psychoactive alkaloid extract slurry and the other
psychoactive alkaloid
extract slurry to obtain a bulk psychoactive alkaloid extract slurry
comprising the
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phosphorylated psychoactive alkaloid and the dephosphorylated psychoactive
alkaloid
in the specific ratio; standardizing the bulk psychoactive alkaloid extract
slurry by
adding thereto a measured quantity of one or more excipients to obtain a
standardized
bulk slurry; and drying the standardized bulk psychoactive alkaloid slurry to
obtain the
psychoactive alkaloid composition, wherein the phosphorylated psychoactive
alkaloid
and the dephosphorylated psychoactive alkaloid are in the specific ratio;
wherein the
specific ratio of phosphorylated psychoactive alkaloid to dephosphorylated
psychoactive alkaloid ranges from 1:1000 to 1000:1.
[502] 153.A psychoactive alkaloid composition comprising: a psychoactive
alkaloid
extract comprising a desired amount of a phosphorylated psychoactive alkaloid
and a
desired amount of a dephosphorylated psychoactive alkaloid, wherein: the
desired
amount of the phosphorylated psychoactive alkaloid is 0-100% by weight of a
total
phosphorylatable psychoactive alkaloid content in the psychoactive alkaloid
extract,
and the desired amount of the dephosphorylated psychoactive alkaloid is the
remainder
of the total phosphorylatable psychoactive alkaloid content in the
psychoactive alkaloid
extract; and one or more excipients.
[503] 154. The composition of embodiment 153, wherein the composition is in
powder form.
[504] 155. The composition of embodiment 153, wherein: the desired amount
of the
phosphorylated psychoactive alkaloid is 0% by weight of the total
phosphorylatable
psychoactive alkaloid content in the psychoactive alkaloid extract, and the
desired
amount of the dephosphorylated psychoactive alkaloid is 100% by weight of the
total
phosphorylatable psychoactive alkaloid content in the psychoactive alkaloid
extract.
[505] 156. The composition of embodiment 153, wherein: the desired amount
of the
phosphorylated psychoactive alkaloid is 100% by weight of the total
phosphorylatable
psychoactive alkaloid content in the psychoactive alkaloid extract, and the
desired
amount of the dephosphorylated psychoactive alkaloid is 0% by weight of the
total
phosphorylatable psychoactive alkaloid content in the psychoactive alkaloid
extract.
[506] 157. The composition of embodiment 153, wherein: the phosphorylated
alkaloid is psilocybin, baeocystin, norbaeocystin, aeruginascin, or any
combination
selected therefrom; and the dephosphorylated alkaloid is psilocin,
norpsilocin, 4-
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hydroxytryptamine, N,N,N-trimethy1-4-hydroxytryptamine, or any combination
selected
therefrom.
[507] 158.A psychoactive alkaloid composition with a specific ratio of a
phosphorylated psychoactive alkaloid and a dephosphorylated psychoactive
alkaloid,
the composition comprising: a psychoactive alkaloid extract having a total
phosphorylatable psychoactive alkaloid content of 100% of a phosphorylated
psychoactive alkaloid; another psychoactive alkaloid extract having a total
phosphorylatable psychoactive alkaloid content of 100% of a dephosphorylated
psychoactive alkaloid; and one or more excipients; wherein the psychoactive
alkaloid
extract and the other psychoactive alkaloid extract are present in a
proportion such that
the specific ratio of phosphorylated psychoactive alkaloid to phosphorylated
psychoactive alkaloid ranges from 1:1000 to 1000:1.
EXAMPLES
[508] In order to further illustrate the present invention, the following
specific
examples are given with the understanding that these examples are intended
only to be
illustrations without serving as a limitation on the scope of the present
invention. All
parameters, dimensions, materials, quantities, and configurations described
herein are
examples only and may be changed depending on the specific embodiment.
Accordingly, the scope of the invention is to be construed in accordance with
the
substance defined by the claims. The process may be scaled up using larger
quantities
and modified apparatus.
[509] Although the examples of the present invention have been formulated
specifically using Psilocybe cubensis as a source to obtain a psychoactive
alkaloid
extract, the extract including psilocybin and psilocin, other sources are
possible. A
person skilled in the art would appreciate that the Psilocybe cubensis can be
readily
substituted by other sources of psychoactive alkaloids to obtain a variety of
other
psychoactive alkaloids having similar properties, such as psilocybin,
psilocin,
baeocystin, norbaeocystin, norpsilocin, aeruginascin, bufotenin, bufotenidine,
4-
hydroxytryptamine, 5-Me0-DMT (5-methoxy-N,N-dimethyltryptamine), N,N-
dimethyltryptamine (DMT), N,N,N-trimethy1-4-hydroxytryptamine, ergine (LSA),
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ergonovine, ergometrine, muscimol, ibotenic acid, lysergic acid
hydroxyethylamide
(LSH), elymoclavine, ergometrinine, chanoclavine, or any combination
therefrom, to
result in compositions with similar efficacy and efficiency as well. For
example,
mushrooms from the genus Conocybe, Cope/and/a, Galerina, Gymnopilus, Inocybe,
Panaeolus, Pholiotina, Pluteus, Psilocybe, or any combination therefrom may be
used.
For example, psilocybe cyanescens and amanita muscaria fungi may be used. For
example, the venom of the toad Inc///us alvarius, the Anadenanthera colubrina
tree or
the Anadenanthera peregrina tree may be used as other sources of psychoactive
alkaloids. Note that the lists of sources and psychoactive alkaloids are
included to
provide examples and are non-exhaustive lists.
Example 1: Preparation of psychoactive alkaloid extract
[510] Referring to FIG. 12, 2.5 kg of fresh Psilocybe cubensis (caps, stems
and
gills) was taken (step 1200) and dried (step 1201) in a forced air oven at 25
C for 5-10
hours. A mass of 140 g of dried biomass was obtained. The dried biomass was
pulverized (step 1202) to a size of 200 mesh with a hammer mill to result in a
dried,
powdered biomass. The dried, powdered biomass was placed in an agitated, heat-
controlled extraction vessel with 5 kg of a hydro-ethanol mixture, with 3
parts ethanol to
1 part water by weight, as a solvent (step 1203). The extraction (step 1204)
was
carried out for 4 hours at a controlled temperature of 70 C to obtain an
extract in the
form of a slurry. The extraction slurry was filtered (step 1205) while it was
hot, and
filtrate A was collected. The filter residue was retained (step 1206) and
placed back
into the extraction vessel, followed by addition (step 1207) of another 5 kg
of 3:1
ethanol:water mixture by weight as a solvent. The extraction was repeated
(step 1208).
The temperature of extraction was again carried out at 70 C for a duration of
4 hours.
The obtained extraction slurry was filtered (step 1209) while hot and filtrate
B was
collected. Filtrate A and filtrate B from the first and second extractions
were mixed
(step 1210). Using a rotary evaporator, the mixture was then partially
concentrated by
evaporation (step 1211) of the solvent from the combined filtrates to provide
a 2.5 liter
volume of psychoactive alkaloid extract solution. Step 1211 is similar to step
1010 of
FIG. 10.
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Example 2: Preparation of psychoactive alkaloid extract
[511] In another example, the process of Example 1 was followed, except
that the
combined filtrates A and B were left to cool to room temperature, and any
precipitate
that formed was filtered out and discarded.
Example 3. Preparation of Anadenanthera peregrina seed extract
[512] 1.00 kg of dried Anadenanthera peregrina seeds were pulverized to a
size of
200 mesh with a grinder. The dried powdered biomass was placed into an
agitated,
heat-controlled vessel with 20 kg of solvent. In this embodiment, the solvent
was a
hydro-ethanol mixture of 4 parts ethanol to 1 part water by weight. The
extraction was
controlled to a constant 70 C, and the time of extraction was 4 hours. The
extraction
slurry was filtered while hot, and the filter residue was placed back into the
extraction
vessel and extracted again with an additional 10 kg of 4:1 ethanol:water
mixture by
weight. The temperature of extraction was again 70 C and the time was 4
hours. The
extraction slurry was filtered while hot and the filtrates from the first and
second
extractions were mixed together. The resulting bulk filtrate was immediately
placed into
an evaporation still and the solvent was removed until the final volume of the
filtrate was
reduced to about 6 liters, which resulted in a solids concentration in the
filtrate of 6.8%.
Example 4: Extraction using a solvent mixture of ethanol (0-100 wt%) and water
[513] Referring to FIG. 3, a process is shown for the extraction of
psychoactive
compounds from Psilocybe cubensis using a general hydro-ethanol solvent. The
solvent may range from a percentage of <1% of ethanol in water to 100%
ethanol.
[514] In step 300, 2.5 kg of raw psilocybin mushrooms from the psilocybe
cubensis
species was provided. In step 310, the raw psilocybe cubensis was dried in a
forced air
oven at 25 C for 10 hours. In step 320, the resulting dried biomass was
ground in a
hammer mill or the equivalent, to particle size of 200 mesh.
[515] In step 330, 5 kg of solvent, having a 0-100% ethanol concentration
was
added to an extraction vessel into which the ground biomass was placed. The
extraction vessel was an agitated, heat-controlled vessel.
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[516] In step 340, the extraction proceeds as the biomass was soaked. The
temperature of the extraction was elevated above room temperature to 70 C.
Temperature and pressure, if applied, were generally selected so that the
solvent does
not boil if elevated temperatures were used. The duration of the extraction
was 4 hours.
[517] In the step 350, the extraction slurry was filtered to remove residue
with
undissolved Psilocybe cubensis from the filtrate. The residue may be treated
with
another extraction step if desired, and if so, the filtrate from the
subsequent step was
combined with the filtrate from the first filtration.
[518] In step 360, the solvent from the filtrate was partially evaporated
using a
rotary evaporator. The resulting concentrated slurry was subjected to a
standardization
process in step 370. The standardized concentrated slurry was then dried using
a
bench-top spray dryer in step 380 to result in a powder with an accurately
determined
concentration by weight of psychoactive alkaloids.
Example 5: Extraction using a solvent mixture of ethanol and water (3:1 wt%)
[519] Referring to FIG. 2, an exemplary detailed process is shown for the
extraction
of psychoactive compounds from Psilocybe cubensis mushrooms using a 75%
ethanol
solvent.
[520] In step 200, 2.5 kg of raw psilocybin mushrooms from the psilocybe
cubensis
species was provided. In step 201, the raw psilocybin mushrooms were dried in
a
forced air oven at 25 C, for 10 hours. The aim was to dry the mushrooms so as
not to
significantly reduce their psychoactive alkaloid concentration. For example,
if too high a
temperature or too long a time at a specific temperature were used, the
alkaloids may
start to decompose. The resulting, dried biomass was 140 g. In step 202, the
dried
biomass was ground using a hammer mill or the equivalent, to a particle size
of 200
mesh.
[521] In step 203, a 5 kg quantity of the 75% (by weight) ethanol solvent,
formed by
mixing 3 parts of ethanol to 1 part of water by weight, was placed in an
extraction
vessel. The dried, ground biomass was also placed in the extraction vessel,
which was
heat-controlled and agitated.
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[522] The extraction proceeds in step 204 as the biomass soaks in the
solvent. The
temperature of the extraction process was 70 C, and the duration of
extraction was 4
hours. The temperature remained constant during the extraction process.
[523] In step 205, the resulting mixture of biomass solids and solvent with
dissolved
extract, was filtered while still hot, i.e. still at 70 C, or slightly lower
due to ambient
cooling. This removed a residue with undissolved psilocybin mushroom
components
from the filtrate. The filter used was a 10 pm sieve. The filtrate from this
step was
filtrate A. In step 206, the residue was retained and placed back into the
extraction
vessel. In step 207, another 5 kg of 75% ethanol was added to the retained
residue.
[524] In step 208, the extraction process of the residue continued at the
same
temperature as for the initial extraction step, i.e. at 70 C, for a time of 4
hours. Again,
the temperature remained constant during the extraction process.
[525] In step 209, the second resulting mixture, of biomass solids and
solvent with
dissolved extract, was filtered to remove the residue of unwanted solid
material. The
filter used was a 10 pm sieve. Note that in other embodiments a differently
sized filter
may be used here or in the prior filtration step, or the liquid may be
decanted from the
residue without filtering. In some embodiments, a centrifuge may be used to
help
separate the liquid from the residue. Filtrate B from the second filtration
process may
have a lower concentration of psychoactive compounds than filtrate A from the
first
filtration step. Filtrates A and B were then mixed in step 210 to result in
bulk filtrate C.
More extract can be obtained by splitting the solvent into two or more batches
and
using each one sequentially to soak the biomass, compared to using a single
volume of
solvent.
[526] The bulk filtrate C was then processed with a rotary evaporator in
step 211 to
remove solvent until the volume of filtrate C is 2.5 liters. At this point,
the reduced
amount of filtrate C was a concentrated slurry, due to the precipitation of
water-
insoluble components, for example.
[527] The volume of 2.5 L was chosen because the mixture now has a low
enough
ethanol content that the carriers can be mixed in. By preferentially removing
ethanol
over water, which occurs naturally during the evaporation, it also gives the
later spray-
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drying step a lower risk of explosion compared to if a 75% ethanol slurry were
sprayed
directly.
[528] In step 212, after some of the solvent has been removed using the
rotary
evaporator, the concentrated slurry was then standardized. The standardization
process uses a titration procedure to determine the concentration of the
psychoactive
alkaloids in the concentrated slurry. The standardization procedure entails
adjusting
the concentration of psychoactive alkaloids the concentrated slurry to a
desired target,
such as 1.00% by dry weight. In this example, 4.7 g of ascorbic acid, 1.9 g of
SiO2 and
47 g of maltodextrin were added to the concentrated slurry.
[529] In step 213, after the standardization process, the standardized
concentrated
slurry was dried using a bench-top spray dryer. This resulted in 100 g of
powdered
psilocybin mushroom extract with a total alkaloid concentration of 1.00% by
weight. As
can be seen, the purity of the extract can be defined as a percentage to a
precision of
two decimal places.
Example 6: Extraction using a buffered solvent mixture of ethanol and water
(3:1 wt%)
(Alkaline solvent)
[530] Referring to FIG. 7, a process is shown for the extraction of
psychoactive
compounds from Psilocybe cubensis mushrooms using a buffered alkaline solvent.
In
step 700, 2.5 kg of raw psilocybin mushrooms from the psilocybe cubensis
species was
provided. In step 701, the raw Psilocybe cubensis was dried in a forced air
oven at
25 C for 10 hours. The dried biomass was 140 g. In step 702, the dried biomass
was
ground in a hammer mill or the equivalent, to a particle size of 200 mesh.
[531] In step 703, 5 liters of solvent was added with the biomass to an
extraction
vessel, which was heat-controlled and agitated. The solvent was a pH-adjusted,
hydro-
ethanol mixture. For its preparation, 200 g of sodium hydroxide pellets were
placed into
a 5 L vessel, with 1.25 L of reverse osmosis water followed by 3.75 L of
ethanol. The
contents were mixed until completely dissolved. The pH of this solution was
between
pH 11 and pH 12.
[532] In step 704, the extraction proceeded. The temperature of the
extraction
process was 30 C, and the duration of the extraction was 4 hours. In step 705,
the
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extraction slurry was filtered to remove residue with undissolved psilocybe
cubensis
from the filtrate. The filtrate from this step was named filtrate A. In step
706, the
residue was retained and placed back in the extraction vessel. In step 707,
another 5 L
of the same solvent was added to the residue. In step 708, the extraction
process of
the residue continued at a temperature of 30 C, for 4 hours. The temperature
remained
constant during the extraction process. In step 709, the second extraction
slurry was
filtered to remove the residue of unwanted solid material. Filtrates A and B
were then
mixed in step 710 to result in bulk filtrate C.
[533] Bulk filtrate C was brought to a pH of 5 with 5M phosphoric acid in
step 711.
The pH-adjusted concentrated slurry was processed with a rotary evaporator in
step
712 to remove solvent until the volume of filtrate C was 2.5 liters. At this
point, the
reduced amount of filtrate C was a concentrated slurry, due to the
precipitation of some
of the psychoactive alkaloids.
[534] In step 713, the concentrated slurry was standardized. In this
example, 4.7 g
of ascorbic acid, 1.9 g of SiO2, and 47 g of maltodextrin were added to the
concentrated
slurry. In step 714, the standardized concentrated slurry was dried using a
bench-top
spray dryer. This resulted in 100 g of powdered psilocybin mushroom extract
with a
total alkaloid concentration of 1.00% by weight.
Example 7: Extraction using a buffered solvent mixture of ethanol and water
(3:1 wt%)
(Acid solvent)
[535] Referring to FIG. 6, a process is shown for the extraction of
psychoactive
compounds from Psilocybe cubensis mushrooms using a buffered acidic solvent.
In
step 600, 2.5 kg of raw psilocybin mushrooms from the Psilocybe cubensis
species was
provided. In step 601, the raw Psilocybe cubensis was dried in a forced air
oven at
25 C for 5-10 hours. The dried biomass was 140 g. In step 602, the dried
biomass
was ground in a hammer mill or the equivalent, to particle size of 200 mesh.
[536] In step 603, 5 L of solvent was added with the dried biomass to an
extraction
vessel, which was heat-controlled and agitated. The solvent was a pH-adjusted,
hydro-
ethanol mixture. For its preparation, 44 g of anhydrous citric acid was placed
into a 5 L
vessel with 1.25 L of reverse osmosis water followed by 3.75 L of ethanol. The
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contents were mixed until completely dissolved. The pH of this solution was
between
pH 1.8 and pH 3.
[537] In step 604, the extraction proceeded. The temperature of the
extraction
process was 30 C, and the duration of the extraction was 4 hours. In step 605,
the
extraction slurry was filtered to remove residue with undissolved psilocybe
cubensis
from the filtrate. The filtrate from this step was named filtrate A. In step
606, the
residue was retained and placed back in the extraction vessel. In step 607,
another 5 L
of the same solvent was added to the residue. In step 608, the extraction
process of
the residue continued at a temperature of 30 C, for 4 hours. The temperature
remained
constant during the extraction process. In step 609, the second extraction
slurry was
filtered to remove the residue of unwanted solid material. Filtrates A and B
were then
mixed in step 610 to result in bulk filtrate C.
[538] Bulk filtrate C was brought to a pH of 5 with 5M sodium hydroxide in
step 611.
The amount of sodium hydroxide depends on the specific mushroom matrix
extracted
and was not possible to predict accurately. The pH-adjusted, concentrated
slurry was
then processed with a rotary evaporator in step 612 to remove solvent until
the volume
of filtrate C was 2.5 liters. At this point, the reduced amount of filtrate C
was a
concentrated slurry, due to the precipitation of some of the psychoactive
alkaloids.
[539] In step 613, the concentrated slurry was standardized. In this
example, 4.7 g
of ascorbic acid, 1.9 g of SiO2, and 47 g of maltodextrin were added to the
concentrated
slurry. In step 614, the standardized concentrated slurry was dried using a
bench-top
spray dryer. This resulted in 100 g of powdered psilocybin mushroom extract
with a
total alkaloid concentration of 1.00% by weight.
Example 8: Extraction using 100% methanol as a solvent
[540] Referring to FIG. 5, a process is shown for the extraction of
psychoactive
compounds from Psilocybe cyanescens mushrooms using 100% methanol as the
solvent.
[541] In step 500, 2.5 kg of raw psilocybin mushrooms from the Psilocybe
cyanescens species was provided. In step 510, the raw Psilocybe cyanescens was
dried in a forced air oven at 25 C for 10 hours. The dried biomass was 140 g.
In step
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520, the dried biomass was ground in a cutting mill or the equivalent, to
particle size of
200 mesh. In step 530, 5 kg of solvent, which was 100% methanol, was added to
an
extraction vessel, which was heat-controlled and agitated. The dries biomass
was also
added to the extraction vessel.
[542] In step 540, the extraction proceeded. The temperature of the
extraction
process was a constant 25 C, and the duration of the extraction was 4 hours. A
pressure of 100 kPa above atmospheric (15 psi) was applied to the mixture of
solvent
and biomass during the extraction. In step 550, the extraction slurry was
filtered to
remove residue with undissolved Psilocybe cyanescens from the filtrate.
[543] The filtrate was then processed with a rotary evaporator in step 560
to
evaporate all the methanol from the filtrate. In this embodiment, all the
solvent was
removed at this stage because methanol was not regarded as safe for human
consumption, and there should be no trace amounts of it remaining in the final
product.
In step 570, 1.25 liters of reverse osmosis water at room temperature was
added to the
solid that is remaining after the evaporation step, to form a concentrated
slurry.
[544] In step 580, the concentrated slurry was standardized. In this
example, 1.84 g
of SiO2 and 46 g of maltodextrin were added to the concentrated slurry. In
step 590,
the standardized concentrated slurry was dried using a bench-top spray dryer.
This
resulted in 95 g of powdered psilocybin mushroom extract with a total alkaloid
concentration of 1.50% by weight.
Example 9: Extraction using 100% water as a solvent
[545] Referring to FIG. 4, a detailed process is shown for the extraction
of
psychoactive compounds Psilocybe cubensis using 100% reverse osmosis water as
the
solvent.
[546] In step 400, 2.5 kg of raw psilocybin mushrooms from the Psilocybe
cubensis
species was provided. In step 401, the raw Psilocybe cubensis was dried in a
forced air
oven at 25 C for 10 hours. The dried biomass was 140 g. Note that the dried
biomass
was the same weight in different examples because the mushrooms were from the
same starting batch. In step 402, the dried biomass was ground in a hammer
mill or the
equivalent, to a particle size of 200 mesh.
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[547] In step 403, 5 liters of solvent, which was 100% reverse osmosis
water, was
placed in an extraction vessel with the dried biomass, which was heat-
controlled and
agitated.
[548] In step 404, the extraction proceeded. The temperature of the
extraction
process was 90 C, and the duration of the extraction was 12 hours. In the step
405,
the extraction slurry was filtered while still hot to remove residue with
undissolved
Psilocybe cubensis from the filtrate. The filtrate from this step was
considered as filtrate
A. In step 406, the residue was retained and placed back in the extraction
vessel. In
step 407, another 5 liters of 100% reverse osmosis water was added to the
residue. In
step 408, the extraction process of the residue continued at a temperature of
90 C, for
hours. The temperature remained constant during the extraction process. In
step
409, the second resulting mixture, of biomass solids and water with dissolved
extract,
was filtered while still hot to remove the residue of unwanted solid material.
Filtrates A
and B are then mixed in step 410 to result in bulk filtrate C.
[549] The bulk filtrate C was then processed with a rotary evaporator in
step 411 to
remove solvent until the volume of filtrate C is 2.5 liters. At this point,
the reduced
amount of filtrate C was a concentrated slurry, due to the precipitation of
some of the
psychoactive alkaloids.
[550] In step 412, after some of the solvent has been removed using the
rotary
evaporator, the concentrated slurry was then standardized. The standardization
process used a titration procedure to determine the concentration of the
psychoactive
alkaloids in the concentrated slurry. The standardization procedure entailed
adjusting
the concentration of the psychoactive alkaloids in the concentrated slurry to
a desired
dry target. In this example, 6.3 g of ascorbic acid, 2.5 g of SiO2 and 63 g of
maltodextrin were added to the concentrated slurry.
[551] In step 413, after the standardization process, the standardized
concentrated
slurry was dried using a bench-top spray dryer. This resulted in 140 g of
powdered
psilocybin mushroom extract with a total alkaloid concentration of 0.50% by
weight.
Example 10. Process with prevention of dephosphorylation
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[552] 2.5 kg of Psilocybe cubensis were dried in a forced air oven at 25 C
for 10
hours to result in 140 g of dried biomass. The dried biomass was then
pulverized to a
size of 200 mesh with a mill.
[553] A basified solvent, i.e. a pH-adjusted, hydro-ethanol mixture, was
prepared.
50 g of sodium hydroxide pellets were placed into a 5 L vessel with 1.25 L of
RO
(reverse osmosis) water followed by addition of 3.75 L of ethanol. The
contents were
mixed until completely dissolved. A basified solvent with 75% Et0H v/v% and a
pH of
13 was obtained.
[554] The dried powdered biomass was placed into an agitated, heat-
controlled
vessel with 5 L of the basified solvent and mixed for extraction of
psychoactive alkaloid.
The extraction was controlled to a constant 75 C, and the time of extraction
was 1 hour.
The extraction slurry was then filtered. Filtration resulted in a filtrate,
i.e. the
psychoactive alkaloid liquid, and a filter residue. The filter residue was
placed back into
the extraction vessel and extracted with an additional 5 L of the basified
solvent. The
temperature of extraction was again 75 C and the time was again 1 hour. The
resulting
extraction slurry was then filtered. The filtrates from the first and second
extractions
were mixed to form 10 L of mixed filtrate. The pH of the mixed filtrate was
then reduced
with 3 M citric acid until a pH of 4.5 was achieved. Immediately after
adjusting the pH,
the mixed filtrate was placed into a rotary evaporator at 50 C and 250 torr,
and the
solvent was partially or completely evaporated to obtain a psychoactive
alkaloid extract
in a slurry or powdered form respectively. Final stages of evaporation were
performed
using a freeze dryer. When dried to a powder, the desired amount of the
phosphorylated psychoactive alkaloid obtained was 0.80% by weight (measured by
HPLC analysis) in the psychoactive alkaloid extract. The desired amount of the
dephosphorylated psychoactive alkaloid obtained was 0.05% by weight in the
psychoactive alkaloid extract.
Example 11. Process for heavily phosphorylated psychoactive alkaloid
composition
[555] The evaporation of the solvent from the mixed filtrate from Example
10 was
paused when the final volume of the filtrate was reduced to 2.5 L. The
resulting slurry
contained 236.9 mg/L psilocybin and 15.12 mg/L psilocin. The obtained
psychoactive
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alkaloid slurry was standardized by the addition of 38.64 g of maltodextrin,
1.12 g of
ascorbic acid, and 2.24 g of silicon dioxide. The standardization was followed
by drying
by freeze drying to yield 112 g of the psychoactive alkaloid composition in
free-flowing
powder form. The composition had a psilocybin content of 0.503% dry wt/wW0 and
a
psilocin content of 0.003% dry wt/wW0. Further, the composition had a total
phosphorylatable psychoactive alkaloid content of 0.506% dry wt/wt%.
Example 12. Process with promotion of dephosphorylation
[556] 2.5 kg of Psilocybe cubensis were dried in a forced air oven at 25 C
for 10
hours to result in 140 g of dried biomass. The dried biomass was then
pulverized to a
size of 200 mesh with a hammer mill.
[557] An acidified solvent, i.e. a pH-adjusted, hydro-ethanol mixture, was
prepared.
144 g of anhydrous citric acid was placed into a 5 L vessel with 1.25 L of RO
water
followed by the addition of 3.75 L of ethanol. The contents were mixed until
completely
dissolved. An acidified solvent with a pH of 2 was obtained.
[558] The dried powdered biomass was placed into an agitated, heat-
controlled
vessel with 5 L of the acidified solvent and mixed for the extraction of
psychoactive
alkaloid. The extraction was controlled to a constant 75 C, and the duration
of
extraction was 1 hour. The extraction slurry was then filtered. Filtration
resulted in a
filtrate, i.e. the psychoactive alkaloid liquid, and a filter residue. The
filter residue was
placed back into the extraction vessel and extracted with an additional 5 L of
the
acidified solvent. The temperature of extraction was again 75 C and the time
was 1
hour. The extraction slurry was filtered. The filtrates from the first and
second
extraction were mixed to form 10 L of mixed filtrate. The pH of the mixed
filtrate was
then increased with 5 M sodium hydroxide until a pH of 4.5 was achieved.
Immediately
after adjusting the pH, the mixed filtrate was placed into a roto-evaporator
at 50 C and
250 torr, and the solvent was partially or completely evaporated to obtain a
psychoactive alkaloid extract. Final stages of evaporation were performed
using a
freeze dryer. When dried to a powder, the desired amount of the phosphorylated
psychoactive alkaloid obtained was 0.00% by weight in the psychoactive
alkaloid
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extract. The desired amount of the dephosphorylated psychoactive alkaloid
obtained
was 0.86% by weight in the psychoactive alkaloid extract.
Example 13. Process for preparation of a psychoactive alkaloid composition
[559] The evaporation of the solvent from the mixed filtrate from Example
12 was
paused until the final volume of the filtrate was reduced to 2.5 L. The
obtained
psychoactive alkaloid slurry had a 241.85 mg/L of psilocin and a 0 mg/L of
psilocybin.
The slurry was standardized by the addition of 47.07 g of maltodextrin, 1.21 g
of
ascorbic acid, and 2.41 g of silicon dioxide. The standardization was followed
by freeze
drying to yield 120.7 g of the psychoactive alkaloid composition in free-
flowing powder
form. The composition had a psilocybin content of 0.00% dry wt/wt% and a
psilocin
content of 0.501% dry wt/wt%. Further, the composition had a total
phosphorylatable
psychoactive alkaloid content of 0.501% dry wt/wt%.
Example 14. Process for preparation of a psychoactive alkaloid composition
containing
a mixture of phosphorylated and dephosphorylated alkaloids
[560] In an alternate method, the slurry from Examples 12 and 13 can be
combined
to form 5 L of slurry containing 118.45 mg/L of psilocybin and 128.49 mg/L of
psilocin.
The slurry was standardized by the addition of 85.70 g of maltodextrin, 2.33 g
of
ascorbic acid, and 4.65 g of silicon dioxide. The standardization was followed
by
lyophilization to yield 232.70 g of the psychoactive alkaloid composition in
the free-
flowing powder form. The composition had a psilocybin content of 0.255% dry
wt/wt%
and a psilocin content of 0.276% dry wt/wt%. Further, the composition had a
total
phosphorylatable psychoactive alkaloid content of 0.502% dry wt/wt%.
Example 15: Purifying the psychoactive alkaloid extract using a non-ionic
macroporous
resin
[561] The pH of the partially concentrated extract of Example 1, which was
an
aqueous extract, was adjusted to pH 4.0 (+/- 0.5) by adding 2 M phosphoric
acid and
centrifuged for 15 minutes at 3000g to remove any solid precipitate. The pH of
4
corresponds to the isoelectric point of psilocybin, and psilocin's polarity is
such that it is
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partitioned onto the resin, thus allowing effective binding of the
psychoactive alkaloids
psilocybin and psilocin to the macroporous resin. Norbaeocystin and baeocystin
are
phosphorylated and behave in the same way as psilocybin. The supernatant
obtained
was loaded onto a column of Amberlite XAD4, a non-ionic macroporous resin
(50.34 g
of dry resin) at a flow rate of 2 bed volumes per hour, to allow components in
the
supernatant to be adsorbed onto the macroporous resin. After all 2.5 L of the
extract
was loaded onto the column of macroporous resin, the column was washed in a
single
pass with 5 bed volumes of reverse osmosis water at room temperature. This was
followed by elution with 5 bed volumes of 5% ethanol (by weight), again at
room
temperature. Finally, the column was washed in a single pass with 5 bed
volumes of
100% ethanol. The elution was performed at room temperature. Each of these
three
fractions was collected separately. The particular order for the washing steps
and the
elution was selected to be in the order of the polarity of the solvents. If
the order were
different, an inferior result may have ensued, such as a lower yield. The
first fraction
using reverse osmosis water removed the most polar compounds from the resin.
The
hydro-ethanol fraction eluted compounds of lesser polarity, and the 100%
ethanol
solvent removed the least polar compounds. Less polar solvents could also be
used to
elute less polar compounds.
[562] The 5% ethanol fraction (i.e. the purified psychoactive alkaloid
solution) was
then concentrated in a rotary evaporator to form 3.90 g of concentrated
aqueous slurry
at 30% solids, containing 195.1 mg of total alkaloids, i.e. psilocybin,
psilocin,
norbaeocystin, and baeocystin. The result was a purified psychoactive alkaloid
slurry
having a total psychoactive alkaloid concentration of 5.00% by weight.
[563] As described below, it is possible to replace the solvent with an
equivalent
weight of excipients to provide a purified extract with a psychoactive
alkaloid content of
5.00% dry wt/wt /0.
Example 16: Purifying the psychoactive alkaloid extract using cation exchange
and
non-ionic macroporous resins
[564] The combination of filtrates of Example 11 was taken as the starting
point.
The pH of the combined filtrate obtained was adjusted to a pH of 3.0 (+1- 0.5)
by adding
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1M HCI. It was then mixed with 200 g of Amberlite MAC-3 H, a strong cationic
ion-
exchange resin in its hydrogen form, to result in a filtrate-resin mixture, in
which
components of the psychoactive alkaloid filtrate were adsorbed onto the cation
exchange resin. The pH of 3 ensured that the psychoactive alkaloid (i.e.
psilocybin)
was in its protonated form, and thus adsorbed onto the cationic exchange resin
efficiently. The filtrate-resin mixture was agitated for 4 hours at room
temperature
(21 C ¨ 25 C) and then filtered. The filtrate was discarded, and the resin was
rinsed
with 2.0 L of 100% Et0H and then 2.0 L of H20 to remove any impurities.
Finally, the
psilocybin/psilocin fraction was eluted with 2.0 L of 2% HCl/80% Et0H, for 4
hours at
room temperature.
[565] The eluted fraction was brought to a pH of 4.0 (i.e. the isoelectric
point of
psilocybin) by adding 2M NaOH. The filtrate was then centrifuged at 3000g to
remove
any solid precipitate. The resultant filtrate, in form of an aqueous solution,
was then
placed into a rotary evaporator and the solvent was removed until the aqueous
solution
reached a volume of 400 mL. The aqueous solution was then again centrifuged
for 15
minutes at 3000g to remove any solid precipitate. The supernatant was loaded
onto a
column of Amberlite XAD4 macroporous resin (45.53 g of dry resin) at a flow
rate of 2
bed volumes per hour. After all the 400 mL of the supernatant was loaded onto
the
column, it was initially washed with 5 bed volumes of reverse osmosis water,
followed
by elution with 5 bed volumes of 5% ethanol (by weight) and then washed with
100%
ethanol. Each of these fractions was collected separately. The 5% ethanol
fraction (i.e.
the purified psychoactive alkaloid solution) was concentrated in a rotary
evaporator to
form 258 mg of solution containing 175 mg of total alkaloids (i.e. psilocybin,
psilocin,
norbaeocystin, and baeocystin). Thus, a purified psychoactive alkaloid slurry
with a
total alkaloid concentration of 68% dry wt/wt% was obtained.
Example 17: Purifying the psychoactive alkaloid extract using anion exchange
and
non-ionic macroporous resins
[566] The combination of filtrates of Example 11 was taken as the starting
point.
The pH of the filtrate combination was adjusted to 9.5 (+/- 0.5) by adding 1 M
NaOH
and then mixed with 150g of Amberchrom 50WX8 strong anionic ion-exchange
resin in
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its hydrogen form to result in a filtrate-resin mixture, in which components
of the
psychoactive alkaloid filtrate were adsorbed onto the anion exchange resin.
The pH of
9.5 (+/- 0.5) ensured that the psilocybin, psilocin, norbaeocystin, and
baeocystin were
deprotonated and had a net negative charge for efficient adsorption onto the
strong
anion exchanger.
[567] The filtrate-resin mixture was agitated for 4 hours and then filtered
out, and
the filtrate was discarded. The resin was rinsed with 2.0 L of 100% Et0H and
then 2.0
L of H20 to remove impurities. Finally, the psilocybin/psilocin fraction was
eluted with
2.0 L of 2% NaCl/80% Et0H for 4 hours.
[568] The eluted fraction was brought to a pH of 4.0 with the addition 2 M
HCI. The
extract was then centrifuged at 3000g to remove any solid precipitate. The
resultant
extract, in from of a solution, was then placed into a rotary evaporator and
the solvent
was removed to result in a volume of 400 mL.
[569] The resultant 400 mL aqueous solution was centrifuged for 15 minutes
at
3000g to remove any solid precipitate. The supernatant was loaded onto a
column of
Amberlite XAD4 macroporous resin (45.53 g of dry resin) at a flow rate of 2
bed
volumes per hour, to allow components of the supernatant to be adsorbed onto
the
macroporous resin. After all 400 mL of supernatant was loaded onto the column,
the
column was initially washed with 5 bed volumes of reverse osmosis water,
followed by
elution with 5 bed volumes of 5% ethanol (by weight) and then a final wash
with 100%
ethanol was performed. Each of these fractions was collected separately. The
5%
ethanol fraction (i.e. the purified psychoactive alkaloid solution) was
concentrated in a
rotary evaporator to form 325 mg of solution containing 175 mg of total
alkaloids (i.e.
psilocybin, psilocin, norbaeocystin, and baeocystin). A purified psychoactive
alkaloid
slurry with a concentration of 54% dry wt/wt% of total alkaloids was therefore
obtained.
Example 18: Purifying the Anadenanthera peregrine seed extract
[570] The aqueous extract with about 6.8% solids, from Example 17, was
adjusted
to pH 4.0 (+/- 0.5) with 2 M phosphoric acid and centrifuged for 15 minutes at
3000g to
remove any solid precipitate. The supernatant was loaded onto a column of
Seplite
LXA17 macroporous resin (54.21 g of dry resin) at a flow rate of 2 bed volumes
per
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hour. After all 6.0 L of the extract was loaded onto the column, it was
initially washed
with 5 bed volumes of reverse osmosis water, followed by a second wash with 5
bed
volumes of 10% ethanol (by weight) and then eluted with 3 bed volumes of 50%
ethanol, and finally the resin was washed with 5 bed volumes of 100% ethanol.
Fewer
bed volumes of solvent were possible in the elution step than in the washing
steps due
to the sharper elution peak. This in turn led to a shorter evaporation time
than if more
bed volumes of the solvent had been used. Each of these fractions was
collected
separately. The 50% ethanol fraction was concentrated in a rotary evaporator
to form
355 g of concentrated aqueous slurry at 30% solids, containing 3.03 g of total
alkaloids.
Example 19: Process for preparing standardized psychoactive alkaloid extract
[571] The 3.90 g of purified psychoactive alkaloid slurry with a
psychoactive alkaloid
concentration of 5.00% by weight that was obtained in Example 15 was taken and
standardized. To achieve this, the concentrated slurry, 0.03 g of SiO2, 0.02 g
of
ascorbic acid and 2.55 g of maltodextrin were added and thoroughly mixed to
result in a
final standardized slurry having a specific concentration of alkaloids. The
final
standardized slurry was then subjected to spray-drying and a final powdered
alkaloid
extract with a 5.00% total psilocybin, psilocin, baeocystin and norbaeocystin
concentration by dry weight was obtained.
Example 20: Process for preparing standardized psychoactive alkaloid extract
from
Anadenanthera peregrina seeds
[572] The aqueous slurry from Example 18 was used as the starting point.
Next,
0.3g of SiO2, 0.15g of citric acid and 4.10 g of maltodextrin were added to
the slurry,
which was thoroughly mixed. The final formulated slurry was then subjected to
spray-
drying to yield a final powdered alkaloid extract with a combined
bufotenin/bufotenidine/5-Me0-DMT concentration of 20.00% by dry weight.
Example 21: Preparation of a non-purified psychoactive alkaloid extract
[573] Fresh Psilocybe cubensis, 2.5 kg, was dried in a forced air oven at
25 C for
5-10 hours, to result in 140 g of dried biomass. The dried biomass was
pulverized to a
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size of 200 mesh with a hammer mill. The dried powdered biomass was then
placed
into an agitated, heat-controlled vessel with 5 kg of solvent. The solvent was
a hydro-
ethanol mixture of 3 parts ethanol to 1 part water by weight. The extraction
was
controlled to a constant 70 C, and the time of extraction was 4 hours.
[574] The extraction slurry was filtered while hot, and the filter residue
was placed
back into the extraction vessel, and extracted with an additional 5 kg of 3:1
ethanol:water mixture by weight. The temperature of extraction was again 70
C, and
the time was 4 hours. The extraction slurry was filtered while hot and the
filtrates from
the first and second extractions were mixed together to obtain a bulk
filtrate.
The bulk filtrate was immediately placed into a rotary evaporator, and the
solvent was
concentrated in the rotary evaporator to obtain 186.6 g of the psychoactive
alkaloid
extract in form of a concentrated aqueous slurry at 30% solids, containing 700
mg of
total alkaloids, which would be a concentration of 1.25% dry wt/wt%, if the
slurry were
to be dried.
Example 22: Preparation of a purified psychoactive alkaloid extract
[575] Fresh Psilocybe cubensis, 2.5 kg, was dried in a forced air oven at
25 C for 5-
hours, resulting in 140 g of dried biomass. The dried biomass was then
pulverized to
a size of 200 mesh with a hammer mill. The dried powdered biomass was placed
into
an agitated, heat-controlled vessel with 5 kg of solvent. The solvent used was
a hydro-
ethanol mixture of 3 parts ethanol to 1 part water by weight. The extraction
was
controlled to a constant 70 C, and the time of extraction was 4 hours.
[576] The extraction slurry was filtered while hot, and the filter residue
was placed
back into the extraction vessel, and extracted with an additional 5 kg of 3:1
ethanol:water mixture by weight. The temperature of extraction was again 70 C
and the
time was 4 hours. The extraction slurry was filtered while hot and the
filtrates from the
first and second extractions were mixed together to obtain a bulk filtrate.
The bulk
filtrate was left to cool, in case any precipitate had formed, the insoluble
material was
filtered out and discarded.
[577] The bulk filtrate's pH was then adjusted to 9.5 (+/- 0.5) with 1 M
NaOH to form
a specific pH psychoactive alkaloid solution. This solution was then mixed
with 150g of
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Amberchrom 50WX8 Strong Anion Exchange resin in its hydrogen form. The
solution
was agitated for 4 hours and then filtered. The filtrate was discarded. The
resin was
rinsed with 2.0 L of 100% Et0H and then 2.0 L of H20. Finally, the psilocybin
fraction
was eluted with 2.0 L of 2% NaCI / 80% Et0H for 4 hours.
[578] The eluted fraction was brought to a pH of 4.0 with 2 M HCI to result
in
another specific pH psychoactive alkaloid solution. This solution was then
centrifuged at
3000g to remove any solid precipitates. The solvent from the solution was then
evaporated in a rotary evaporator to result in a volume of solvent evaporated
was 400
mL.
[579] This solution was again centrifuged for 15 minutes at 3000g to remove
any
solid precipitate. The supernatant was loaded onto a column of Amberlite XAD4
macroporous resin (45.53 g of dry resin) at a flow rate of 2 bed volumes per
hour. All
400 mL of the extract was loaded onto the column and washed with 5 bed volumes
of
reverse osmosis water. The washing step was followed by elution with 5 bed
volumes
with 5% ethanol (by weight). A final washing was carried out with 100%
ethanol. Each
of these fractions was collected separately. The 5% ethanol fraction was
collected and
concentrated in a rotary evaporator to obtain 1.143 g of 30% liquid slurry
containing
175 mg of total alkaloids, a concentration of 54% dry wt/wt%.
Example 23: Preparation of a psychoactive alkaloid composition with a non-
purified
psychoactive alkaloid extract
The psychoactive alkaloid extract obtained in Example 12 was mixed with 2.8 g
of
silicon dioxide (flow agent), 0.140 g of ascorbic acid (preservative), and
81.06 g of
tapioca maltodextrin (carrier). The final formulated slurry was then subjected
to spray-
drying and 140 g of the standardized powdered composition was produced with
the
desired specific amount of psychoactive alkaloid. The total
psilocybin/psilocin
concentration by dry weight was 0.5% in this composition. The exact weight
percentages of the components in the composition are depicted Table 1.
Example 24: Preparation of a psychoactive alkaloid composition with a purified
psychoactive alkaloid extract
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[580] The purified psychoactive alkaloid extract obtained in Example 22 was
mixed
with 5.85 mg of ascorbic acid (preservative) and 822 mg of rice maltodextrin
(carrier).
The final formulated slurry was then subjected to lyophilization and 1.171 g
of the
standardized powdered composition was produced. The total psilocybin/psilocin
concentration by dry weight was 15.01% in the composition. The exact weight
percentages of the components in the composition are depicted Table 1.
Example 25: Process for preparing standardized psychoactive alkaloid extract
[581] A purified psychoactive alkaloid solution resulting from resin
treatment after
extraction from 140 g of dried Psilocybe cubensis was concentrated in a rotary
evaporator to form 3.90 g of concentrated aqueous slurry at 30% solids,
containing
195.1 mg of total psychoactive alkaloids. The slurry, with a psychoactive
alkaloid
concentration of 5.00% by weight, was mixed with 0.03 g of SiO2, 0.02 g of
ascorbic
acid and 2.55 g of maltodextrin. This standardized slurry was then subjected
to spray-
drying, and a final powdered alkaloid extract with a 5.00% total psilocybin,
psilocin,
baeocystin and norbaeocystin concentration by dry weight was obtained.
Example 26: Process for preparing standardized psychoactive alkaloid extract
[582] An extract from 140 g dried Psilocybe cubensis mushrooms using a 75%
ethanol solvent resulted in a concentrated slurry, for which the alkaloid
content was
2.16 g and the total solid content was 46.4g. To the slurry, 4.7 g of ascorbic
acid, 1.9 g
of SiO2 and 47 g of maltodextrin were added. After spray drying, this resulted
in 100 g
of powdered psychedelic mushroom extract with a total alkaloid concentration
of 1.00%
by weight.
Example 27: Process for preparing standardized psychoactive alkaloid extract
[583] Psychoactive compounds were extracted from 140 g of dried Psilocybe
cubensis using 100% reverse osmosis water as the solvent. Water was evaporated
to
result in a concentrated slurry, for which the alkaloid content was 1.82 g and
the total
solid content was 68.18 g. In this example, 6.3 g of ascorbic acid, 2.5 g of
SiO2 and 63
g of maltodextrin are added to the concentrated slurry, which was then dried.
This
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resulted in 140 g of powdered psilocybin mushroom extract with a total
alkaloid
concentration of 0.50% by weight.
Example 28: Process for preparing standardized psychoactive alkaloid extract
[584] An extraction of psychoactive compounds from 140 g dried Psilocybe
cyanescens mushrooms was performed using 100% methanol as the solvent. The
extraction slurry was filtered to remove residue with undissolved Psilocybe
cyanescens
from the filtrate. All the methanol was evaporated from the filtrate, then
1.25 liters of
reverse osmosis water at room temperature was added to the remaining solid to
form a
slurry, for which the alkaloid content was 2.87 g and total solid content was
47.14 g.
Next, 1.84 g of SiO2 and 46 g of maltodextrin were added to the slurry, which
was then
dried. This resulted in 95 g of powdered psychedelic mushroom extract with a
total
alkaloid concentration of 1.50% by weight.
Example 29 - 3.7: Process for preparing standardized psychoactive alkaloid
extract
[585] An extract obtained according to Example 22 was used as the starting
point.
Compared to Example 24, greater amounts of preservative (351 mg), flow agent
(351
mg) and carrier (16.515 g) were added to the composition. This resulted in a
standardization of the amount psychoactive alkaloid in the composition to
1.00%
instead of 15.01%.
Example 30: Process for preparing standardized psychoactive alkaloid extract
[586] A purified psychoactive alkaloid solution resulting from resin
treatment after
extraction from 140g of dried Psilocybe cubensis was concentrated in a rotary
evaporator to form 3.90 g of concentrated aqueous slurry at 30% solids,
containing
195.1 mg of total psychoactive alkaloids. Compared to Example 25, greater
amounts of
preservative (0.49 g), flow agent (0.39 g), and carrier (18.43 g) were added
to the
composition. This resulted in a standardization of the amount psychoactive
alkaloid in
the composition to 1.00% instead of 5.00%.
Example 31: Process for preparing standardized psychoactive alkaloid extract
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[587] A
purified psychoactive alkaloid solution was obtained after multiple cation
exchange resin treatments following extraction from 140 g of dried Psilocybe
cubensis.
Silicon dioxide, maltodextrin and ascorbic acid were added to form a
composition
standardized to 60.00%.
Example 32: Process for preparing standardized psychoactive alkaloid extract
[588] This is as Example 31, except that the only excipient that was added
was
preservative (ascorbic acid). This resulted in a standardization of the
psychoactive
alkaloid content of the composition to 75.00%.
[589] The exemplary compositions obtained are depicted in Table 1.
Compositions
of Examples 23 and 26-28 are compositions with a psychoactive alkaloid extract
that
has not been purified. Compositions of Examples 24, 25, and 29-32 are
compositions
with a purified psychoactive alkaloid extract.
Table 1. Compositions of Examples 23-32
Alk aloid Alkaloid
Extract Preservative Flow Agent Carrier Total Amount in
Amount in
Ex. Mass Mass Mass Mass Mass
Standardized
Extract
(dry %) (dry %) (dry %) (dry %) (dry %)
Composition
wt/wt%) (wt/wt%)
23 40.0 0.1 2.0 57.9 100.0 1.25 0.50
24 29.3 0.5 0.0 70.2 100.0 51.23 15.01
25 31.0 0.5 0.8 67.6 100.0 16.12 5.00
26 46.4 4.7 1.9 47.0 100.0 2.16 1.00
27 48.7 4.5 1.8 45.0 100.0 1.03 0.50
28 49.6 0.0 1.9 48.4 100.0 3.02 1.50
29 2.0 2.0 2.0 94.0 100.0 51.23 1.00
30 6.2 2.5 2.0 89.3 100.0 16.12 1.00
31 79.6 5.0 1.0 14.4 100.0 75.22 60.00
32 99.7 0.3 0.0 0.0 100.0 75.22 75.00
[590] In the columns, the extract mass (dry %), preservative mass (dry %),
flow
agent mass (dry %), and carrier mass (dry %) are the dry weight percentages of
the
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psychoactive alkaloid extract, preservative, flow agent and carrier in the
standardized
composition respectively. The total mass (dry %) is the total dry weight
percentage of
the standardized composition. The alkaloid amount in the extract (wt/wt`)/0)
is the dry
weight percentage of the psychoactive alkaloid in the extract, as if the
extract were in its
dried state. Note that it is possible for the extract to remain in the slurry
state as the
excipients are added. The alkaloid amount in the standardized composition
(wt/wt%) is
the dry weight percentage of the psychoactive alkaloid in the final
composition. It can
be seen that a wide variability in extract concentration from different
batches can be
standardized to the same concentration in the composition, e.g., by looking at
examples
26, 29, and 30.
Example 33: 95% Methanol/5% acetic acid (Psilocybin-rich extract)
[591]
14.5 kg of fresh Psilocybe cubensis were dried in a forced air oven at 30 C
for
48 hours, resulting in 1.45 kg of dried biomass mushroom fruiting body. The
content of
psilocybin in the biomass was 0.523 % by dry weight, resulting in 7.25 g of
psilocybin
available for extraction. The dried biomass was reduced to a size of 200 mesh
with a
cutting mill. The dried powdered biomass was placed into an agitated, heat-
controlled
vessel with 58 L of solvent (40 L/kg). FIG. 24 shows a chart for solvent to
solid ratio
optimization. A S:S (solvent:solid) ratio of 20-30 L/kg can achieve >90%
alkaloid yield
over 3 extractions while the optimal condition was chosen as 40-50 L/kg,
achieving
>90% alkaloid yield in only two extractions, reducing the amount of solvent
waste, time,
and energy during the evaporation step. In this embodiment, the solvent was
acidified
methanol (5% acetic acid/ 95% anhydrous methanol v/v%). It is noteworthy that
methanol works very well, and acidified methanol works 10-15% better. Both are
acceptable extraction methods for psilocybin. The extraction was controlled to
a
constant 25 C temperature and was under atmospheric pressure. The extraction
was
carried out under these conditions for 30 minutes, after which the extraction
slurry was
filtered through a 5 pm stainless steel filter. The filtrate was placed into
another vessel
and put aside. FIG. 25 shows a graph of time and temperature optimization,
which
indicates that the temperature increases the extraction efficiency, but also
shows
degradation when extended beyond 20-30 minutes. Given that, at scale, the
increased
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complexity of elevating the temperature of the extraction vessel would
introduce a
significant warming and cooling time, it was decided to select the extraction
temperature of 20 C, and keep the extended extraction time at 30-50 minutes
to avoid
degradation of the alkaloids. The now-dry filter cake was again placed into
the
extraction vessel and an additional 58 liters of extraction solvent was added
to the
vessel. The extraction was again carried out under the same conditions for 30
minutes.
The slurry was then filtered and combined to create the pooled filtrate. The
pooled
filtrate was placed into a rotary-evaporator, and the methanol was evaporated
until the
volume was reduced to around 5.8 L, forming a concentrated solution. 5.8 L was
roughly 5% of the pooled filtrate volume, which was entirely acetic acid at
this point and
around pH 2.4. Basically, it is required to remove all of the methanol for
further
purification. It may be also possible to take this solution and evaporate it
fully to
dryness, add excipient and have a shelf-stable low purity extract (0.5- 2.0%
alkaloid
content by weight). It is possible to have this material continue onto
purification. The
content of psilocybin in this concentrated solution was 1.18 g/L, and the
yield was
94.2%. The dry mass yield at this stage was 44.23%. Components that are still
present in the extract at this point are: small chain
carbohydrates/polysaccharides, free
sugars, polyphenols, alkaloids, some glycoproteins, ergothioneine,
tocopherols,
ergosterols, fats. Many of these components are targeted for removal with
purification.
Components that are present in the mushroom that are left behind in the
biomass:
proteins, large carbohydrates/polysaccharides, B-glucans.
[592] The concentrated solution was diluted with RO water to 50 L (1.28 %
dry
mass concentration). The aqueous extract was then adjusted to pH 4.0 (+/- 0.5)
with 2
M sodium hydroxide and filtered through a 5pm stainless steel filter to remove
any solid
precipitate. It is very important to have the extract be at pH 4.0 before
application to the
adsorbent resin, because pH 4 is the isoelectric point of psilocybin, and it
is also the
maximum stability pH for psilocin. The supernatant was loaded onto a column of
Amberlite XAD4 macroporous resin (5000 mL of hydrated resin, -1.39 mg
psilocybin/mL of hydrated resin) at a flow rate of 2 bed volumes per hour.
Optimization
of breakthrough and determination of capacity is shown in FIG. 26 (see also
Table 2).
After all 50 L of extract is loaded onto the column, it was washed with 3 bed
volumes of
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reverse osmosis water at a flow rate of 2 BV/h, followed by elution with 5 bed
volumes
of 15% ethanol (by weight) and then finally washed with 100% ethanol. Each of
these
fractions was collected separately. FIG. 27 shows a graph of optimized
recovery of
psilocybin on XAD4 with 1 pass. Desorption with 15% ethanol resulted in 99.2%
recovery of psilocybin while retaining only 2.5% of the dry mass in the same
fraction.
This resulted in a ¨40X increase in concentration over the extract and a 45%
dry
wt/wt% content of psilocybin in the first pass purified extract. In one
embodiment, it is
preferred to stop here and bring in the excipients for stabilization and
standardization to
a dry powder. If it is intended to go higher and purer, it is preferred to
expose the
concentrated extract to another pass on the XAD4 or to an !solute SCX resin
or
antisolvent addition, or liquid/liquid extraction. The 15% ethanol fraction
contained 6.79
g of psilocybin and 16.03 g of dry mass, resulting in an extract of 42.35%
psilocybin by
weight. The 15% ethanol fraction was then concentrated in a rotary evaporator
to form
53.45 g of concentrated aqueous slurry at 30% solids.
[593] 1.09 g of SiO2, 1.36 g of ascorbic acid, 1.36 g of citric acid, 17.24
g of
maltodextrin, and 17.24g of mannitol were added to the concentrated aqueous
slurry,
and it was thoroughly mixed. The final formulated slurry was then subjected to
lyophilization, and the final powdered alkaloid extract concentration was 12.5
% total
psilocybin concentration and less than 0.4 % psilocin by dry weight. SiO2 and
maltodextrin were added as flowability enhancers. Mannitol is a cryoprotectant
(allowing for efficient freeze-drying) and bulking agent. Ascorbic acid is an
antioxidant
(allowing protection from oxidation by first oxidizing itself), and citric
acid is a chelating
agent that may impart increased bioavailability and pH buffering once inside
the
stomach. This composition has been developed and has shown 9 months of shelf
stability (see Table 3).
Table 2. XAD4 Psilocybin Capacity at Three Different Flow Rates
2 BV/h 4 BV/h 6 BV/h
Capacity (mg Alkaloids) 765 mg 600 mg 463 mg
mL Resin 550 550 550
mg Alkaloids/mL XAD4 1.39 1.09 0.84
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Table 3. Individual Time-Point Data of Psilocybin, Psilocin and Moisture
Content of
PYEX-FP-200820
Months Psilocybin (WIWI%) Psilocin (wt/wt%) Moisture Content
M.)
0 13.99 0.12 0.91 0.02 7.23
1 13.89 0.11 0.90 0.02 6.94
2 14_00 0.19 0.91 0.02 6.77
3 13_91 0.28 092 0.03 6.93
4 13.81 0.22 0.92 0.02 7.14
13.86 0,10 0.92 0.04 7.22
6 13.89 0,07 0.92 0.04 6.87
7 13.95 0,09 0.93 0.03 6.9
8 13.85 0,16 0.96 0.02 7.31
9 13.93 0,17 0.93 0.04 6.93
Example 34: 0.15 M Citric acid/water (Psilocin extract)
[594] 15.7 kg of fresh Psilocybe cubensis mushrooms were dried in a forced
air
oven at 30 C for 48 hours, resulting in 1.57 kg of dried biomass mushroom
fruiting
body. The content of psilocybin in the biomass was 0.523 A by dry weight,
resulting in
8.21 g of psilocybin available to convert to 5.90 g psilocin
(stoichiometrically). The dried
biomass was reduced to a size of 200 mesh with a cutting mill. The dried
powdered
biomass was placed into an agitated, heat-controlled vessel with 78.5 L of
solvent (50
L/kg). FIG. 29 shows a chart of solvent to solid ratio optimization for water.
The
optimal is 50 L/kg which was the lowest S:S ratio that can obtain >90%
psilocin yield in
two extractions. In this embodiment, the solvent was acidified water (0.15 M
citric acid,
pH 2.0). The extraction was controlled to a constant 25 C temperature and was
under
atmospheric pressure. The extraction was carried out under these conditions
for 60
minutes, and the extraction slurry was filtered through a 5 pm stainless steel
filter. The
filtrate was placed into another vessel and put aside. The now-dry filter cake
was again
placed into the extraction vessel, and an additional 78.5 L of extraction
solvent was
added to the vessel. The extraction was again carried out under the same
conditions
for 60 minutes. The slurry was then filtered and combined to create the pooled
filtrate.
The pooled filtrate had a content of psilocin of 0.034 g/L, and the yield was
91.23%.
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The dry mass yield at this stage was 68.92%. The dry mass yield was so high
because
of the citric acid content.
[595] The filtrate was directly loaded onto a column of AmberliteTM XAD4
macroporous resin (2700 mL of hydrated resin, ¨1.98 mg psilocin/mL of hydrated
resin)
at a flow rate of 2 bed volumes per hour (FIG. 30).
[596] After all 157 L of extract was loaded onto the column, it was washed
with 3
bed volumes of reverse osmosis water at a flow rate of 2 BV/h, followed by
elution with
bed volumes of 15% ethanol (by weight) and then finally washed with 100%
ethanol.
Each of these fractions was collected separately. The 15% ethanol fraction
contained
5.11 g of psilocin and 14.94 g of dry mass, resulting in an extract of 34.20%
psilocin by
weight. The 15% ethanol fraction was then concentrated in a rotary evaporator
to form
49.80 g of concentrated aqueous slurry at 30% solids.
[597] 0.82 g of SiO2, 1.64 g of ascorbic acid, 0.41 g of citric acid, 11.54
g of potato
starch, and 11.54 g of mannitol was added to the slurry and thoroughly mixed.
The
final formulated slurry was then subjected to lyophilization, and the final
powdered
alkaloid extract concentration was 12.5 A total psilocin concentration and no
psilocybin
was present in the formulation.
Table 4. XAD4 psilocybin capacity at three different flow rates
2 BV/h 4 BV/h 6 BV/h
Capacity (mg Alkaloids) 811 mg 678 mg 595 mg
mL Resin 410 550 550
mg Alkaloids/mL XAD4 1.98 1.65 1.45
Example 35. Psilocybe Cubensis extraction, purification, and stabilization
rationale
[598] The indole alkaloids present in psilocybe mushrooms can be separated
into
two categories, the phosphorylated pro-drugs (psilocybin, norbaeocystin,
baeocystin
and aeruginascin) and their dephosphorylated active constituents (psilocin, 4-
hydroxytryptamine, norpsilocin and 4-hydroxy-N,N,N-trimethyltryptamine
respectively).
When these molecules are consumed (whether in the raw mushroom, purified
extracts,
or synthetic preparations of the compounds) the phosphorylated analogues are
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enzymatically metabolized by the human gut, liver, and kidneys to the active
dephosphorylated forms. When considering a standardized extract composition,
there
are benefits to controlling whether the extraction conditions will favor
producing the
dephosphorylated or phosphorylated forms.
[599] The dephosphorylated indole alkaloids are well known to be unstable.
Internal experiments have also quantified and substantiated these findings
under
aqueous conditions across the pH spectrum (Tables 5 and 6).
Table 5. pH Stability of purified psilocybin over time.
Recovery (%)
Psiloc bin T=0 T=4h T=16h
pH 1 98.43 97.23 98.32
pH 2 97.56 9.5.72 95.84
01.3 90.46 102.32 102.08
pH 4 95.60 102..69 102.20
pH 5 94.01 9.3.89 93.23
.pH 6 103.30 102.30 10T.09
pH 71 95.72 100.21 98. yo
pH 8 98.41 98.44 97.31
pH 9 97.92 95.87 95.46
pH 10 1102.32 103.19 102.60
pli 11 105.38 105.50 104.68
pH 12 100.68 100.24 96.33
Table 6. pH Stability of purified psilocin over time.
Recovery r/o)
Psilocin TO T=4h T1 61i
pH 2 1102..75 77.08 57.80
PH 3 90.83 90.83 72.02
pH 4 93.58 99.08 .81.47
pH 5 34.40 69.27 59.45
pH 6 95.41 74.22. 82.66
pH T 96.33 74.40 48.72
PH 8 90.83 74.40 48.62
Phi 9 78.07 4.3.49 0.00
pH 10 88.99 42.39 20.46
pH 11 86.24 39.17 0.00
pH 12 88.07 22.29 10.37
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[600] While the dephosphorylation reaction is inevitable when consumed by
humans, it also happens rapidly within the Psilocybe mushrooms themselves as a
defense mechanism. The "bluing reaction" that occurs when the fresh mushrooms
are
bruised (while harvesting or handling) is a result of enzymatic
dephosphorylation of
psilocybin to psilocin, followed by enzymatic oxidation and dimerization of
psilocin to
semiquinoid dimers and polymers (FIG. 32A) (Lenz 2020). In the present
disclosure, a
method that either promotes or inhibits various steps of this degradation
cascade and
uses them has been developed to be advantageous during the extraction of
indole
alkaloids from psilocybe mushrooms.
[601] Psilocybin and psilocin are soluble in methanol and water and are not
soluble
in isopropanol, ethanol and acetone. Experiments were performed to determine
the
best solvent compositions for extraction (FIGS. 32B and 32C; Table 7)
Table 7. Dry weight concentration of each extract produced from each
composition
(dried)
wt/wrio Alkaloid Concentration In Dry Extract
%Solvent
Methanol Ethanol lsopropanol Acetone
0 0 066 0.066 0 066 0.066
20 C.131 0.190 0 180 0.060
40 0.184 0.233 0 586 0.075
60 0.765 1.031 0.988 0.068
80 0 836 1.283 1 233 0.000
100 1.313 0.399 0.780 0 000
[602] Methanol was shown to be the superior solvent of choice for
extracting these
alkaloids in general; the optimal composition was 100% anhydrous methanol
which
resulted in a near 100% extracted yield of psilocybin and psilocin. Any
percentage of
water in the extraction was shown to decrease yield of both analytes,
indicating that the
mushroom biomass should be as dry as possible so as to introduce a minimal
amount
of water. It is likely that any amount of water present in the extraction
solvent allows
phosphatase to act upon the phosphorylated alkaloids, converting them to their
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dephosphorylated counterparts. In the solvent compositions that have low total
yield,
laccase enzyme is also actively oxidizing psilocin to their semiquinoid
breakdown
products.
[603] 80% ethanol was also an effective solvent, however, it was observed
that
while the total alkaloid yield was high (>90%), some psilocybin was converted
to
psilocin during the extraction.
[604] Since psilocybin and psilocin are soluble in water, experiments were
conducted to determine if pH modulation could be beneficial in the extraction
process
(FIG. 32D).
[605] High pH water was shown to extract and prevent degradation of
phosphorylated alkaloids, while low pH was shown to extract and
dephosphorylate
psilocybin but prevent laccase oxidation activity. This process was validated
and can
be used reliably to control the dephosphorylation process. On first look, it
would appear
that a drawback is that the use of high and low pH appears to be an "all or
none"
process where there is no modulation of the dephosphorylation cascade.
[606] However, combining the information gained from the water extraction
pH
study with the superior ability of methanol to extract these alkaloids,
experiments were
performed to see how this could be used to improve the process. An interesting
result
was revealed by varying the methanol/water ratio while fixing the pH with 5%
acetic
acid (¨pH 2.5, range 1-4) (FIG. 32E).
[607] It was evident that the oxidation of psilocin during extraction was
only weakly
observed at the 0% methanol ratio, validating the previous experimental data
with low
pH water extraction. Either protonation of the 4-hydroxy position on the
psilocin
molecule was inhibiting laccase's ability to catalyze the oxidation and free
radical
formation, or the enzyme was denatured and not active at this pH. Gradually,
as the
methanol concentration in the extraction solvent is increased, a higher
proportion of the
total alkaloid content is retained as the phosphorylated form, indicating that
methanol is
inhibiting or drastically slowing phosphatase's ability to cleave the
phosphate bond.
Another possibility is that at a certain concentration, methanol is no longer
able to
extract or solvate the phosphatase enzyme, leaving it behind in the solid
residue during
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PSU004-CADIV1
extraction. This process can be used to modulate the
phosphorylated/dephosphorylated content of the extract.
[608] Macroporous resin adsorption/desorption was chosen as a simple and
efficient purification platform to concentrate the total alkaloid content
since the
concentration factor from extraction alone was underwhelming. Once the proper
composition of alkaloids is determined (Table 8), a number of purification
permutations
can be utilized (Table 9).
Table 8. Summary of extraction processes and conditions used
EXTRACTION
Goal: Most preferred Less Preferred
-High pH adjusted
Water
-Acidified Anhydrous -80%
Methanol Ethanol/Water
Phosphorylated alkaloids left - Anhydrous Methanol -
High pH Adjusted
intact (Psilocybin, Baeocystin, 80%
Norbaeocystin, Aeruginascin) Ethanol/Water
-50-70%
-Acidified Water Ethanol/Water
Dephosphorylated Alkaloids
-Acidified 0-20%
(Psilocin)
methanol/water
-Combine extracts from
Specific Ratio of Psilocybin to different methods AFTER
Psilocin purification at specific ratio
-acidified 20-80% Methanol
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Date Recue/Date Received 2022-07-25
PSU004-CADIV1
Table 9. Summary of purification processes and condition
PURIFICATION
Goal: Most preferred Less Preferred
-SCX, SAX or other
Resin
Repeated XAD4 column -Antisolvent addition
Highest Purity Psilocybin
runs with acetone
-Liquid/liquid
extraction
XAD4 + SCX or
XAD4 column run +
Highest Purity Psilocin LXA817 antisolvent or liquid
extraction
Partial Purification of
Single XAD4 run
psilocybin or psilocin
[609] Throughout the description, specific details have been set forth in
order to
provide a more thorough understanding of the invention. However, the invention
may
be practised without these particulars. In other instances, well known
elements have
not been shown or described in detail and repetitions of steps and features
have been
omitted to avoid unnecessarily obscuring the invention. Accordingly, the
specification
and drawings are to be regarded in an illustrative, rather than a restrictive,
sense.
[610] All parameters, dimensions, materials, quantities and configurations
described
herein are examples only and may be changed depending on the specific
embodiment.
Numbers and percentages are given to the nearest significant figure. For
example,
10% includes the range between exactly 9.5% and exactly 10.5%. Accordingly,
the
scope of the invention is to be construed in accordance with the substance
defined by
the claims. The process may be scaled up using larger quantities and a
modified
apparatus.
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Date Recue/Date Received 2022-07-25
