Note: Descriptions are shown in the official language in which they were submitted.
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DELIVERY OF FUNCTIONAL COMPOUNDS
CROSS-REFERENCE TO RELATED APPLICATIONS
[00011 This application claims benefit of United States Provisional
Application Number
61/422,439, filed December 13, 2010, and is a continuation-in-part of United
States Patent
Application Number 12/479,444, filed June 5, 2009, both of which are hereby
incorporated by
reference in their entirety herein.
FIELD
100021 This application relates to a modified functional ingredient which
is micro-
encapsulated by an enteric matrix and methods for making the same. More
particularly, the
modified functional ingredient is rnicroencapsulated in an aqueous environment
which is
substantially free of organic solvents.
BACKGROUND
[00031 Enteric delivery of functional materials in food applications has
been limited.
Enteric delivery systems are commonly utilized when the functional materials
or medicaments
are known to be sensitive to certain conditions such that they become less
effective or if the
functional materials cause problems for the user, such as stomach problems
with aspirin.
Generally, enteric delivery, as most common in pharmaceutical practice, is
accomplished using
coated tablets and gel capsules. However, those particular delivery methods
are not well suited
for food applications. In particular, neither tablets nor capsules are sized
to be integrated into
most existing food products.
100041 An alternative process for enteric delivery is microencapsulation.
Micro-
encapsulation is generally performed using specialized equipment or in an
environment
including organic solvents. These methods require additional capital
expenditures and the use
of additional materials, such as the organic solvents, which may or may not be
usable in
subsequent microencapsulation cycles. As a result, the process of
microencapsulation requires
investments in both equipment and organic solvent procurement and disposal.
100051 One issue with microencapsulation is the recovery rate, or
microencapsulation
efficiency of the process. Generally, a certain significant percentage of the
material to be
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microencapsulated is not captured. The uncaptured material may be recovered
for reuse,
recycled, or a percentage of the uncaptured material remains adhered to the
outer surface of the
microencapsulated particulates.
[00061 As a result, the product tends to have a taste profile associated
with the uncaptured
material, which is often undesirable. This is particularly true when the
uncaptured material
includes oxidizable triglycerides such as unsaturated and polyunsaturated
lipids, oxidizable
flavors and essential oils, or other organic compounds that may naturally have
undesirable taste
and/or flavor.
SUMMARY
[00071 The composition includes a functional ingredient microencapsulated
in an enteric
matrix, such as described in US. Patent Application Serial No. 12/479,454,
which is
incorporated by reference in its entirety herein. The enteric matrix includes
one or more food
grade polymer(s), and the functional ingredient includes a modified functional
ingredient
[00081 In one embodiment, the functional ingredient is homogeneously
dispersed
throughout the enteric matrix material. In another embodiment, the functional
ingredient
includes at least about 30 percent modified compounds. In one form, the
modified compound
includes at least one of a salt, glycoside, complex or ester of an essential
oil, such as linalool and
thymol.
100091 In one aspect, a method of microencapsulating a modified active or
functional
ingredient is provided. The method includes agitating or mixing water, an
enteric matrix
material, and an emulsifier to form a combination at a pH that maintains
complete dissolution
of the enteric matrix material being utilind. In one approach, the combination
is substantially
free of organic solvents. A functional ingredient including a modified portion
is added to the
combination and homogenized to create a fine, stable emulsion. The emulsion is
then treated
with an acid and/or other cross-linking or precipitating agents, depending on
the enteric matrix
material being used, such as calcium, under controlled mixing conditions and
in an amount and
at a rate effective to form a particulate precipitate. Further, the functional
ingredient is
homogeneously dispersed throughout the precipitate and with an improved
microencapsulation efficiency of the functional ingredient.
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BRIET DESCRIPTION OF THE DRAWINGS
[0010] FIG. I illustrates a method for microencapsulating a functional
ingredient;
[0011] FIG. 2 is a chart comparing the microencapsulation efficiency
between two
experiments, with one experiment including a functional ingredient that does
not contain at
least 30 percent esters, the second experiment including a functional
ingredient that includes at
least 30 percent esters of linalool and thymol;
[0012] FIG. 3 is a table showing the known empirical formula, solubility in
water, vapor
pressure, partition coefficient and a ratio comparing the affinity for oil to
water ratio of various
esters;
[00131 FIG. 4 is a graph illustrating the percentage release of the various
components of a
functional ingredient which does not include esters with an enteric matrix
comprised of
95 percent shellac and 5 percent zein;
10014] FIG. 5 is a graph illustrating the percentage release of the various
components of a
functional ingredient including esters with an enteric matrix comprised of 95
percent shellac
and 5 percent zein;
[0015] FIG. 6 is a graph illustrating the percentage release of the various
components of a
functional ingredient comprising linalyl acetate within an alginate/ shellac
enteric matrix in a
digestion model simulating stomach and small intestine conditions; and
[00161 FIG. 7 is a graph illustrating the percentage release of the various
components of a
functional ingredient comprising linalyl butyrate within an alginate/ shellac
enteric matrix in a
digestion model simulating stomach and small intestine conditions.
DETAILED DESCRIFI __________________________ ION
[00171 Disclosed is a method of microencapsulating a modified functional
ingredient and a
non.-active carrier(s) in an enteric matrix that minimizes release of the
functional ingredient
prior to dissolution in the intestine. Further, the funchonal ingredients can
have undesirable
taste or flavor profiles. In some cases, modifying the functional ingredient
can mask or change
the undesirable tastes and/or flavors while retaining the proper ratios of the
functional
ingredients ensuring bioavailability and efficacy.
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100181 In one form, the functional ingredient to be microencapsulated can
include modified
forms of essential oils, such as thymol and linalool. Modifications to the
functional ingredient
may include a variety of forms that modify the perceived taste and/or
organoleptic properties
of the functional ingredient. For example, the modification may cause a change
to the flavor
and/or taste threshold of the functional ingredient. In one form, the
modification causes a
change to the volatility and/or vapor pressure of the modified functional
ingredient with
respect to the non-modified, parent form of the functional ingredient. In
particular, the
organoleptic properties of the modified form may include a higher taste
threshold. As a result,
in some cases a modified functional ingredient on the surface of the enteric
matrix may produce
a less undesirable flavor profile than the presence of unmodified functional
ingredients.
Further, the modifications may include salt, glycoside, complex and/or
esterification of the
functional ingredient.
[00191 When ingested and released in the intestinal tract, the modified
form of the
functional ingredient reverts back, at least in part, into the parent form and
provides the same
functional benefits as if the parent functional ingredient was
microencapsulated and consumed.
In one form, the modified functional ingredient hydrolyzes from the modified
form back into
the parent, non-modified form of the functional ingredient during digestion.
Further, the
modified forms of the functional ingredient can provide additional benefits as
will be discussed
further below.
[00201 The modified form may make up varying percents of the overall
functional
ingredient composition. For example, the modified form comprises at least 10%
of the
functional ingredient composition. In another example, the modified form
comprises at least
30% of the functional ingredient composition. In yet another example, the
modified form
comprises at least 50% of the functional ingredient composition.
[00211 Further, where the modified functional group includes an ester,
other benefits are
realized. Esters are generally known to produce less undesirable flavors, and
therefore any
flavors produced would not result in a wholly undesirable organoleptic flavor
profile. Further,
due to the low water solubility and/or increased hydrophobicity of the
modified functional
ingredients including an ester, particularly in reference to the parent non-
esterified functional
ingredients, the below described method can result in a higher
microencapsulation efficiency, as
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can be shown by a higher payload and retention rate, than has been recognized
in the absence
of e.sterified functional ingredients.
[00221 Examples of the use of the product created by the methods described
herein are
aimed at delivery in a powdered soft drink (P5D) beverage. Other exemplary
uses of the
product include other food products, such as biscuits, bars, ice cream,
snacks, instant meals and
the like.
[0023] A method for microencapsulating a functional ingredient is generally
described in
FIG, 1. Enteric delivery within a food matrix is achieved by the formation of
matrix particles
with the dispersed portion being that of the functional ingredient, such as an
essential oil blend
with diluent triglycerides, and the matrix portion is that of food grade
enteric polymers, such as
shellac, zein, calcium alginate, denatured whey protein, caseinate and any and
all food-grade
enteric polymers.
[0024] As shown in FIG, 1, water, an enteric matrix material and an
emulsifier are mixed or
agitated until the enteric matrix material and emulsifier are fully dispersed
in the water 1.00.
Generally, the emulsifier and enteric matrix material can be added to the
water together or
separately, with either being added first. The pH is maintained at a level
sufficient that allows
for complete solubilization of the enteric material. As an example, for use of
shellac, zein or
combinations thereof, the pH of the dispersion is generally between about 7.2
and 9,0. In some
embodiments, a basic material, such as sodium, potassium or ammonium
hydroxide, can be
added to the dispersion to raise the pH, such as within a range from about 7.2
to about 1.2.0,
preferably 8.0 to 11.3, to guarantee and maintain complete dissolution of the
enteric polymers
without the use of organic solvents,
[0025] As used herein, "agitation" or "agitated" refers to the use of a top
entering mixer
with impeller or a rotor/stator mixing device operating at a speed of less
than 10,000 RPM.
100261 As used herein, "substantially free of organic solvent" refers to an
amount of added
organic solvent, such as isopropanol or ethanol or any other organic solvent
less than the
amount required to enable solubility of the enteric material under the
processing conditions. In.
one approach, the amount of added organic solvent is less than about 0,1
percent by weight of
the combination of water, emulsifier and enteric material.
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[00271 In one embodiment, the water is deionized water.
[00281 The enteric matrix material used herein is any food grade enteric
polymer, or a
combination of two or more food grade enteric polymers. Preferably, the
enteric matrix
material is shellac, zein, calcium alginate, caseinate or combinations
thereof. Other food grade
enteric polymers include denatured whey protein. In one approach, the prepared
enteric matrix
material does not contain any organic solvents.
[00291 The emulsifier described herein may be any food grade emulsifier. In
preferred
embodiments, the emulsifier is polysorbate, polyglycerol ester, sucrose
stearate, sucrose esters,
proteins, lecithins or combinations thereof. More particularly, the emulsifier
is preferably a
sucrose ester due to the creation of the smaller and most uniformly dispersed
oil droplets
within the later created emulsion.
[00301 Generally, water comprises about 50 percent to about 95 percent of
the dispersion by
weight and preferably from about 70 to about 95 percent, and more preferably
from about 80 to
about 90 percent. The emulsifier generally comprises less than about 5 percent
of the dispersion
by weight, preferably from about 0.01 to about 1 percent by weight, and more
preferably about
0.01 to about 0.1 percent by weight of the dispersion. In one approach, the
enteric matrix
material ranges from about 1 percent to about 10 percent by weight, in other
approaches from
about 4 to about 7 percent, and yet in other approaches from about 5 percent
to 6 percent by
weight of the dispersion.
[00311 Upon forming the dispersion, a functional ingredient, having at
least a portion
thereof modified, and a non-active carrier is added 200 and agitated 300 to
provide a coarse
emulsion having a droplet size of more than about 10 micrometers. After the
coarse emulsion is
formed, the coarse emulsion is homogenized 300 to create a fine, stable
emulsion. In one
approach, the fine, stable emulsion has a droplet size of less than about 10
micrometers. Within
the fine emulsion, the functional ingredient and non-active carrier are
homogeneously
dispersed in the form of fine droplets throughout. In one approach, the
combination of the
functional ingredient and non-active carrier is added in amount ranging from
about 2 to about
7 percent of the emulsion by weight. In other approaches, the combination of
the functional
ingredient and non-active carrier is added in an amount ranging from about 3
to about
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6 percent of the emulsion by weight. The emulsion includes from about 60 to
about 95 percent
water.
[00321 As used herein, "homogenization" or "homogenized" refers to mixing
at a speed
greater than about 10,000 RPM, such as the use of a rotor/stator mixing device
or at a lower
mixing speed of an elevated pressure, such as a valve homogenizer operating at
a pressure of
about 500 psi to about 10,000 psi.
[00331 In one approach, the functional ingredient may include chemically
modified
compounds of essential oils. As an example, the functional ingredient includes
chemically
modified compounds of thyrnol and linalool. For example, glycosides, salts
and/or complexes
of thymol and/or linalool may be used. In one form, thymyl and linalyl acetate
may be used.
Further, other fatty acid esters may be used, such as octanoate. Other
acceptable chemically
modified compounds can be used, such as butyrates, lactates, cin.namates and
pyruvates. In
another example, the functional ingredient includes alpha-pinene, para-cymene,
thymyl esters
or salts and linaly1 esters or salts. As discussed in the examples below, one
exemplary blend
includes, by weight, about 18 percent canola oil, about 8 percent alpha-
pinene, about 39 percent
para-cymene, about 5 percent linalool acetate and about 27 percent thymyl
acetate.
[00341 The modified functional ingredient portion can comprise from about 1
to about
99 percent of the functional ingredient by weight. In some approaches, the
modified functional
ingredient can include from at least about 10 percent of the functional
ingredient by weight and,
in other approaches, about 30 percent by weight. In another embodiment, the
modified
functional ingredient can include from about 25 to about 65 percent of the
functional ingredient
by weight.
[00351 In one embodiment, the blend of non-active carrier and functional
ingredient can
include, by weight, about 15 to about 30 percent canola oil, about 1 to about
10 percent
alpha-pinene, about 5 to about 25 percent para-cymene, about 5 to about 20
percent linalyl ester
and about 20 to about 60 percent thymyl ester. In other approaches, the blend
of non-active
carrier and functional ingredient can include, by weight, about 20 to about 25
percent canola oil,
about 2 to about 7 percent alpha-pinene, about 10 to about 20 percent para-
cymene, about 7 to
about 15 percent linalyi ester and about 35 to about 50 percent thymyl ester.
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[0036] Generally, and in one approach, an esterified form of a functional
ingredient, such as
thymol and linalool, can be used regardless of the chemical or biochemical
reaction approach
for its preparation. For example, any ester formed between the hydroxyl
group(s) of a terpene
and an organic or inorganic oxoacid (containing single or multiple oxoacid
groups) may be used
as the functional ingredient. The volatility and vapor pressure of the
modified functional
ingredient can impact the perception of the ingredient. For example, the size
of the ester group
may be modified to provide a desired volatility. In one approach, the ester
linakage is at least
an ethyl ester. It should be noted that larger ester groups may also be used.
100371 By one approach, the selected esterified form may have increased
functionality due
to an increased rate of hydrolysis over the parent form after ingestion and
release from the
enteric matrix in an intestinal tract. Esters may be obtained from natural
sources or synthesized
using any suitable chemical or biochemical reactions between functional
ingredients, such as
thymol and linalool, and organic or inorganic oxoacids that yield esters.
Suitable oxoacids may
include carboxylic acid, amino acids, phosphoric acid, sulfuric acid, and
nitric acid. The
hydroxyl group can be derived from a homogenous source (e.g. thymol) or mixed
source
(thymol and linalool). Exemplary monocarboxylic acids include, but are not
limited to, acetic,
propionic, butyric, pentanoic, hexanoic, decanoic, stearic, lactic, cinnamic,
pyruvk, benzoic, and
gluconic acids. Exemplary dicarboxylk acids include, but are not limited to,
oxalic, malonic,
maleic, fumaric, tartaric, succinic, glutaric, glucaric, adipic, pimelic,
suberic, azelaic, and sebacic
acids. Exemplary tricarboxylic acids, include, but are not limited to, citric
and isocibic acids.
Other exemplary esters that may be formed by reactions of terpenes with
oxoacids include
dithymol succinate, dithymol adipate, and dithymol sebacate.
[0038] in another form, the modified functional ingredient can include an
ester formed,
regardless of chemical or biochemical reaction approach for its preparation,
between terpene
esters and other esters. In particular, the functional group can be formed
using
transesterification. For example, the functional group can include an ester
formed by reacting
thymol acetate with methyl octanoate or tripalmitin.
10039] Alternatively, it is anticipated that the functional ingredient can
include other
modified compounds. In one form, the modified functional group can include any
glycoside
formed by chemical or biochemical reaction between the hydroxyl group(s) of a
terpene and a
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single sugar group (monosaccharide) or several sugar groups (oligosaccharide).
For example,
thymol and/or linalool glycosides can be the modified functional ingredient.
The sugar group
can include any glycoside with the glycone portion composed of mono-, di- tri-
, and/or
polysaccharides of any kind and the aglycone portion being any hydroxy-terpene
(e.g. thymol,
linalool). The sugar group can also include reducing sugars and/or non-
reducing sugars.
Exemplary sugars include, but are not limited to, glucose, fructose,
galactose, ribose, sucrose,
mannose, maltose, lactose, and cellobiose.
100401 In another form, it is anticipated that the functional group can
include any ionic or
nonionic salt or complex formed involving a hydroxy-terpene and another
chemical species.
For example, thymol and linalool salts or complexes can be the modified
functional ingredient.
One example may be sodium and/or potassium chloride. In another form, the
modified
functional ingredient may include thymol salts that do not have fixed
stoichiometries. For
example, thymol salts may be prepared as partial or mixed salts having
different ratios of
cations and thymol comprising one or more specific cations (Na+, K+, Mg++,
etc.) to prepare
solid complexes. The solidified complexes may or may not be obtained in
crystalline form. The
salt or complex may be formed by any suitable method, but in some cases is
formed by a
chemical reaction or association between one or more hydroxy-terpene and one
or more
alkaline reagent. Exemplary alkaline reagents may include, but are not limited
to, alkaline
hydroxide, oxide, or carbonate. The salt or complex can include any alkali
metal, alkaline earth
metal, or transition metal element, or combination thereof. Suitable salts or
complex for use in
foods may include sodium, potassium, lithium, calcium, magnesium, iron,
manganese, zinc,
and aluminum. Other exemplary salts include any mono-, di-, or trivalent salt
of thymol,
including sodium thymolate (e.g. sodium thymoxide) and any mono-, di-, or
trivalent salt of
phenol, including calcium phenoxide.
100411 Furthermore, the functional ingredient may include various forms of
modification
that are combined. For example, a portion of the modified functional
ingredient composition
may include one or more of salts, glycosides, complexes and esterified forms
of one or more
essential oils.
[00421 In some approaches, the functional ingredient may include a mixture
of any essential
oils as described herein. Further, the functional ingredient can be selected
to include materials
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which are desired to be released enterically. As an example, the functional
ingredient can
include compositions described in U.S. Patent Publication No. 2008/0145462 to
Enan. For
example, the functional ingredient includes about 25 to about 35 percent by
weight
para-cymene, about 1 to about 10 percent by weight linalool, about 1 to about
10 percent by
weight alpha-pinene, about 35 to about 45 percent by weight thymol, and about
20 to about
30 percent by weight soybean oil.
[00431 In some approaches, the functional ingredient described herein can
include
compounds which possess functional properties, such as anti-parasitic, anti-
protozoan, and
anti-fungal. In one embodiment, the organic compounds further include alpha-
pinene and
para-cymene.
[0044] In another embodiment, organic compounds are blended with a non-
active carrier
such as a lipid, fatty acid, triglyceride or food grade oil, such as soybean
oil or canola oil.
[0045] The volatile nature of some of the functional ingredients leads to a
very low
threshold value for olfactory perception, resulting in an undesirable
flavor/taste in the existing
beverage/food. In an effort to mask the flavor of the functional ingredients,
the process
described herein includes the inclusion of a far less water soluble form of a
functional
ingredient, such as an esterified form or other modified form of the
functional ingredient (e.g.
esterified forms of thymol and linalool) as well as processes to remove
unencapsulated material
from the final ingredient. For example, esters generally have a less negative
impact on the
taste/flavor of the food system than their respective parent compounds.
Additionally,
complexes, glycosides and salts of essential oils have a less negative impact
on the taste/flavor
of the food system than their respective parent compounds.
[0046] Due to the low water solubility and/or increased hydrophobicity, an
ester may have
higher microencapsulation efficiency, such as described above, than non-
esterified parent
compounds, such as thymol and linalool. Preferably, the efficiency increases
about 50 to about
200 percent over the efficiency observed when using non-esterffied functional
ingredients, more
preferably about 100 to about 150 percent. Further, esters have a higher
olfactory perception
threshold than the parent compounds, such that amount of esters necessary to
be perceived is
more than the amount of non-esterified thymol and linalool.
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[00471 Turning back to the method of FIG. I, the emulsion is then
precipitated by titration
with an acid or with a cross-linking or precipitating agent 400. During
precipitation, the
emulsion can be subjected to agitation. In one embodiment, the emulsion is
titratecl with a
solution of about 1 to about 5 percent calcium chloride and about 1 to about 5
percent citric acid.
In another embodimentõ the emulsion is titrated with acid in an amount
effective to decrease the
pH down to the isoelectric point of the polymer present in the emulsion, such
as a pH of
about 7, causing phase separation and inducing precipitation of the enteric
matrix out of
solution with the functional ingredient being microencapsuiated therein, thus
creating a slurry
of an aqueous solution and precipitate. The precipitated slurry has a particle
size from about 1
to about 1000 micrometers, in some cases about 10 to about 500,0 micrometers,
and more
preferably from about 75 to about 250 micrometers. In other approaches,
precipitation occurs at
a pH ranging from about 3 to about 6, or further between a pH ranging from
about 3.8 to
about 4.6.
100481 While not wishing to be limited by theory, it is believed that as
the pH of the
emulsion drops down to the isoelectric point, the particles of enteric
materialõ such as shellac
and Zei11, may cross-link to like particles or to one another to form a
matrix, the functional
ingredient and non-active carrier being microencapsulated within the matrix.
As a result, of the
cross-linking, the functional ingredient is homogenously dispersed throughout
the matrix. The
matrix further provides a seal for the functional ingredient. As a result, the
impact of the
functional ingredient on the organoleptic qualities of the finished powder is
correlated to any
functional ingredient remaining adhered to the outer surface of the enteric
matrix.
[00491 The acid used can include any food grade acid. In one embodiment,
the acid is citric
acid.
[0050] As noted above, the composition of the enteric matrix material
affects the dissolution
rate and the protection provided by the enteric matrix.
[00511 To reclaim the precipitate, the slurry is filtered 500, washed 600
and dried 700. In
one embodiment, the slurry is filtered, the resultant slurry cake is then
washed and refiltered
prior to drying.
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[00521 In some instances, there will remain at least marginal
unencapsulated material on
the outer surface of particulate precipitate. With the desire to mask
compounds with low
perception thresholds, the u.nencapsulated material generally is reduced to
levels below
perception in the final product matrix and/or solution. In some cases, the
functional ingredient
on the outer surface of the particulate precipitate is less than about 1
percent by weight of the
final product.
[00531 In some cases, a surface oil remover is added to the slurry after
filtering to aid in
removing residual surface oil from the precipitate, as described in U.S,
Patent Application Serial
No, 12/479,433, which is incorporated by reference in its entirety herein.
Further, the surface oil
remover can also be added prior to the refiltering step.
100541 After the precipitate has been filtered and washed, the precipitate
is dried 700 to
form a powder. Drying can be conducted. such that the powder has a moisture.
content of less
than about 10 percent, in other approaches to a moisture content of about 2 to
about 6 percent,
and in yet other approaches to about 3 to about 5 percent.
[00551 Further, the powder can be pulverized to reduce the particle size of
the powder
precipitate, and then further dried to a moisture content of less than about 5
percent, such as
with a fluidized bed dryer. The resultant particles have a particle size
ranging from about 1 to
about 1000 micrometers, in some cases from about 10 to about 500 micrometers,
and in other
cases from about 75 to about 250 micrometers.
[00561 When drying the powder, the temperature should be maintained between
about
25 C to about 70 C, and in some cases about 35 C to about 65 C. During other
processing steps,
it may be suitable to maintain the temperature between about 4 C to about 40
C, in other cases
about 4 C to about 30 C, and in yet other cases from about 15 C to about 28 C,
100571 As will be discussed further below, and as shown in FIG. 2, the
inclusion of modified
functional ingredients, such as thymyl and linaly1 esters, may result in an
increased payload of
the functional ingredients in the final product. The payload refers to the
weight percentage of
the functional ingredients in relation to the final product. Therefore, an
increase in payload
corresponds to an increase in functional ingredient per a given amount of
enteric matrix. In
some cases, the payload of the functional ingredients ranges from about 5 to
about 50 percent.
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The increased payload can be attributed, at least in part, to the decreased
water solubility
and/or increased hydrophobicity of a functional ingredient including modified
functional
ingredients, such as esters, in comparison to a functional ingredient which is
not modified.
Further, in some cases, large volumes of water may be used in the disclosed
process. A
modified functional ingredient, such as esters of thymol and linalool, would
reduce leaching of
the functional ingredient due to the decreased water solubility and/or
increased
hydrophobicity. The ability to limit losses during processing allows for
control over the final
ratio of functional compounds in the food system, as shown to be important in
US. Patent
Application Publication No. US 2008/0145462 Al, (Enan, E. et al.)
Additionally, some
functional ingredients, such as thymol, are crystalline solids at room
temperature (about 20 to
about 25 C). The substitution with a modified functional ingredient, such as
thymyl acetate,
which is a liquid at room temperature (about 20 to about 25 C) would allow for
easier
processing since a crystalline solid ingredient would no longer have to be
solubilized in a liquid
organic solvent, such as by dissolving thymol or other solid ingredient in
triglyceride oil,
ethanol, or other liquid, before preparation of the emulsion.
100581 Advantages and embodiments of the methods described herein are
further
illustrated by the following Examples. However, the particular conditions,
processing schemes,
materials, and amounts thereof recited in these Examples, as well as other
conditions and
details, should not be contrasted to unduly limit this method. All percentages
are by weight
unless otherwise indicated.
100591 Example 1: The Evaluation and Selection of Emulsifiers
[00601 Various emulsifiers were combined with deionized water at about 60 C
to produce
about 2% solutions. The resulting solutions were combined with an essential
oil blend
composition (about 4% alpha-pinene, about 30% para-cymene, about 7% linalool,
about 35%
thymol and about 24% soybean oil) in a 50:50 weight ratio with deionind water
to create an oil-
in-water emulsion. The emulsifiers evaluated were Glycosperse 5-20 KFG (Lonza;
Fairlawn,
NJ), Polyaldo 10-1-0 KFG (Lanza; Fairlawn, NJ), Aldosperse MS-20 KFG (Loma;
Fairlawn, NJ),
Polyaldo 10-2-P KFG (Lonza; Fairlawn, NJ), Ryoto sugar ester (5-1570,
Mitsubishi-Kagaku Food
Corp.; Tokyo, Japan), Precept 8120 (Central Soya; Fort Wayne, IN) and sodium
caseinate
(Alanate-180, New Zealand Dairy Board; Wellington, New Zealand). The sucrose
ester (5-1570)
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was identified as a good emulsifier due to the creation of the smallest and
most uniformly
dispersed oil droplets within the emulsion. The emulsion created with the
sucrose ester also
showed good stability after 24 hours storage at room temperature (about 20 to
about 25"C).
[0061] Example 2: Microencapsulation of a Functional ingredient Containing
Some
Esterified Components in an About 75% Shellac / About 25% Zein Matrix
[00621 About 2400 g of distilled, deionized water (D.I.H20) was added to a
beaker mixed
using StedFast Stirrer SL1200 (Yarnato Scientific; Tokyo, Japan) with a 4-
pronged impeller blade
at speed setting between 5 and 6. About 37.5 g of Jet Milled zein (F4000,
Freeman Industries;
Tuckahoeõ NY) powder was added to the beaker and mixed until uniformly
dispersed. Next,
about 10% NaOH aqueous solution was added until the pH reached about 11.3. The
zein-water
mixture was agitated until the zein powder was completely dissolved and the
solution was
translucent. Next, about 450 g of the pre-made shellac (Temuss #594; Ajax,
Ontario, Canada) in
ammonium hydroxide solution (25% solids) was added and mixed for about 5 to
about
minutes. Finally, about 1.4 g of sucrose ste.arate, 5-1570 (Mitsubishi-Kagaku
Food Corp.;
Tokyo, Japan) was added while mixing, such as for about 5 to about 10 minutes
until
homogeneous mixing has occurred.
[00631 Nextõ about 80 g of the essential oil blend (about 18.8% canola oil,
about 8.6% alpha-
pinene, about 39.8% para-cymene, about 5.4% !Maly' acetate and about 27.4%
thymyl acetate)
was added and mixed for about 5 to about 10 minutes. Using the PowerGen 7001)
(Thermo
Fisher Scientific; Waltham, MA), the mixture was homogenized by blending for
about 4 minutes
at about 15,000 rpm, and then at about 20,000 rpm for about an additional
minute to create the
stable emulsion. Acid titration of the emulsion using about 3% citric acid
solution followed
using Master Flex pump (Barnan.t Corp.; Barrington, IL) at the highest speed
setting with
moderate overhead mixing until the pH reached about 3.8, thereby creating a
slurry.
[00641 About 10 g of 5i02 AB-D (PPG Industries; Pittsburg, PA) was added to
the slurry
and continued to be mixed for about 20 to about 30 minutes. The mixture was
filtered using a
#200 mesh (75 micrometer) screen to produce a cake. In a separate, clean 4000
ml plastic beaker,
about 2000 g of D.1. H2O and about 2.5 g of Si 02 AB-D was mixed to create a
solution. The cake
was resuspended in this solution and mixed for about 3 to about 5 minutes. The
mixture was
filtered using a #200 mesh screen to produce a second cake.
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10065] In a separate, clean 4000 ml plastic beaker, about 2000 g of DJ. H20
and. about 2,5 g
of 5102 AB-.D was mixed to create another solution. The second cake was
resuspended again
and mixed for about 3 to about 5 minutes. The filtrate was pressed using
cheese cloth to
remove the extra moisture. The filtrate was then spread evenly on large tray
on top of a cookie
sheet for overnight drying, uncovered and at room temperature (about 20 to
about 25 C).
[00661 The dried filtrate particles were ground using a Magic Bullet MB1001
(Sino Link
International Trading Co.; Zhejiang, China). Particles between about 75 to
about 250 micro-
meters were separated using #60 and #200 mesh sieves. The moisture content was
measured
using the CEM Smart System 5 (CEM Corp,; Matthews, NC). To reduce the moisture
content to
less than about 6 percent, the filtrate was dried in a Uni-Glatt fluid bed
dryer (Glatt Air
Techniques; Ramsey, NJ) at about 40 C, checking about every 5 minutes. As a
result, the final
product had a moisture content of less than about 6 percent. The fraction was
sifted by passing
through a #60 mesh screen and collected on a #200 mesh screen, thereby
producing particles
having a size of less than about 250 micrometers and greater than about 75
micrometers, The
composition, payload, and surface oil of the resultant product are illustrated
in the table below.
alpha- para-. Unalyl. Thyinyl Cancila Essential
, Pinene Cymene Linalool Acetate Thymol Acetate Oil Oil
Total
. ,
' wt % wt % wt % wt % wt % wt % wt % wt
% wt %
Payload 057, , 1.3 <0.1 ' 1.8 0.12, 9.5
5.0 13.4 18.4
Surface
oils 0.09 0.31 <0.001 0.34 <0.002 1,6 3,0 2.3
5.3
100671 Example 3: Microencapsulation of a Functional Ingredient Containing
Some
Esterified Components in an About 75% Shellac / About 25% Zein Matrix at a
Pilot Plant Scale
[0068] About 12 kg of water and about 7.5 g of sucrose stearate (S4570,
Mitsubishi-Kagaku
Food Corp.; Tokyo, japan) was added to a mixing tank and mixed for about Ito
about 2
minutes. Then about 2,25 kg of pre-made shellac solution (Temuss #594; Ajax,
Ontario,
Canada) in ammonium hydroxide solution (about 25% solids) was added, followed
by about
187,5 g zein powder (F4000, Freeman Industries; Tuckahoe, NY). About 10%
sodium hydroxide
solution was metered in until pH reached about 11.3 (to solubilize the zein).
Once the zeth and
shellac are completely in solution, about 400 g of essential oil blend (about
13% canola oil, about
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10% alpha-pinene, about 25% para-cymeneõ about 12% linaly1 acetate., and about
40% thymyl
acetate) was added. The mix was agitated for about 5 minutes to create an
emulsion.
[0069] The emulsion was titrated with about 3 percent citric acid solution
until the pH
reached about 3.9. About 75 g of Si02 AB-D (PPG industries; Pittsburg, PA) was
added and
mixed for about 20 to about 30 minutes. The slurry was then filtered using a
200 mesh
(75 micrometer) screen. The filter cake on top of the screen was suspended in
about 9.1 kg of
water with about 50 g Si02 AB-D and mixed for approximately 5 minutes, and
then re-filtered
on the #200 mesh screen. The rinsing was repeated one more time, and the final
filter cake was
spread on a tray for drying overnight at room temperature (about 20 to about
250C). The
following day, the product was pulverized in a Waring blender (Waring Lab
Science;
Torrington, CT), and dried in a UniGlatt fluid bed (Glatt Air Techniques;
Ramsey, NI) at 40 C,
and sieved to a desired size (about 75 - about 250 micrometers), The payload
and surface oil of
the resultant product are illustrated in the table below.
Essential Oil Total
Sample Description wt % wt %
plant scale w/ ester compounds Surface
__________________________________ oils 3.8 7.2
[00701 Example 4: Microencapsulation of a Functional Ingredient Containing
Esterified
Components with Increased Oil Loading in a Shellac / Zein Matrix Containing
Whey Protein as
an Emulsifier
[0071] About 2400 g of D.I. H20 was added to a beaker and mixed using
SteilFast Stirrer
51,1200 (Yamato Scientific; Tokyo, Japan) with a 4-pronged impeller blade at
speed setting
between 5 and 6. About 32.5 g of Jet Milled zein powder (F4000, Freeman
Industries; Tuckahoe,
NY) was added to the beaker and mixed until uniformly dispersed. About 10%
NaOH solution
was added until the pH reached about 11.3 and mixed until the zein powder was
completely
dissolved and the solution was translucent. About 20 g of BiPro WPI (Da visco
Foods
International; Eden Prairie, MN) powder was next added and mixed until the
powder was
completely dissolved. Next, about 390 g of the pre-made shellac solution with
about 25 percent
solids (Temuss #594; Ajax, Ontario, Canada) containing ammonium hydroxide was
added and
mixed for about 5 to about 10 minutes until .the solution was homogeneous.
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PM] About 151.4g of the essential oil blend (about 13% canola oil, about
10%
alpha-pinene, about 25% para-cymene, about 12% linalyl acetate, and 40% about
thymyl
acetate) was added and mixed for about 5 to about 10 minutes. Using the
PowerGen 7000
(Thermo Fisher Scientific; Waltham, MA), the mixture was homogenized for about
4 minutes at
about 15,000 rpm and then at an increased speed of about 20,000 rpm for about
1 additional
minute to create a stable emulsion. About 3% citric acid solution was titrated
into the emulsion
using Master Flex pump until the pH reached about 3.8, thereby creating a
slurry.
[00731 About 10 g of Si02 AB-D (PPG Industries; Pittsburg, PA) was added to
the slurry
and mixed for about 20 to about 30 minutes. The mixture was filtered using a
#200 mesh screen
to produce a filtrate cake. In a separate, dean 4000 ml plastic beaker, about
2000 g of D.I. H20
was added and mixed using a StedFast Stirrer. The pH was adjusted to 3.8 41-
about 0.2 by
adding about 3% citric acid solution. About 1 g sucrose stearate, S-1570
(Mitsubishi-Kagaku
Food Corp.; Tokyo, Japan) was added and mixed until completely dissolved,
followed by an
addition of about 2.5 g of Si02 AB-D. The cake was resuspended in this
solution and mixed for
about 3 to about 5 minutes. The mixture was filtered using #200 mesh (75
micrometer) screen to
produce a second filtrate cake. In a separate, clean 4000 ml plastic beaker,
about 2000 g of D.I.
H20 was mixed and pH adjusted to 3.8 +/- about 0.2 by adding about 3.0 percent
citric acid
solution. About 1 g sucrose stearate was added and mixed until completely
dissolved, followed
by an addition of about 2.5 g of Si 02 AB-D. The cake was resuspended in this
solution and
mixed for about 3 to about 5 minutes. The mixture was filtered again using
#200 mesh (75
micrometer) screen to produce a third filtrate cake.
[0074] The resulting filtrate was pressed using cheese cloth to reduce
moisture content. The
filtrate was spread evenly on a large tray on top of a cookie sheet to dry
overnight, uncovered
and at room temperature (about 20 to about 25 C).
[0075] The resultant particles were ground using a Magic Bullet MB1001
(Sino Link
International Trading Co.; Zhejiang, China). The particles sized less than
about 250
micrometers were separated from the rest using a #60 mesh screen. To reduce
the moisture
content to less than about 6 percent, the filtrate was dried in a Uni-Glatt
fluid bed dryer (Glatt
Air Techniques; Ramsey, NJ) at about 40C, checking approximately every 5
minutes. As a
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result, the final product had a moisture content of less than about 6 percent.
The composition,
payload, and surface oil of the resultant product are illustrated in the table
below.
alpha- para- Linaly1 Thyrnyl Canal a Essential Oil
Pinerie Cyniene Una loot Acetate Thymol Acetate Oil Total Total
wt % wt % wt % wt wt % wt % wt % wt % wt
Payload 1,12 21 <0.10 21 <0.10 113 5.7 17.9 23.5
Surface
oils 0.014 0.094 <0.001 0.11 <0.002 0.55 2.8
0.76 3.6
100761 Example 5: Microencapsulation of a Functional Ingredient Containing
Some
Esterified Components in a Matrix Containing About 48% Alginate, About 40%
Shellac and
About 12% Whey Protein as an Emulsifier
[00771 About 2,1 g of sodium alginate (LILV-1,3G, Kimica Corp; Tokyo,
Japan) and about
11.2 g of sodium alginate (I-3G-150, Kimica Corp; Tokyo, Japan) were added to
about 551 g of
water. Under agitation, about 70 g of an about 10% BiPro (Davisco Foods
International; Eden
Prairie, MN) whey protein isolate solution and about 56 g of the pre-made
shellac (Temuss
#594; Ajax, Ontario, Canada) in ammonium hydroxide solution, (about 25%
solids) was added.
Next, the essential oil blend (about 17,4% canola oil, about 6.6% alplia-
pinene, about 26.5%
para-cymeneõ about 7,9% linalyl acetate, and about 41,4% thymyl acetate) was
added and mixed
until a homogenous emulsion was formed with target droplet size of about 4 to
about
7 micrometers, verified with the Horiba particle size analyzer (Floriba
Industries; Irvine, CA),
The solution was then atomized, creating moderately small drops between about
25 to about
300 micrometers, into an aqueous solution bath containing about 2.5% CaC12 and
about 2.5%
citric acid. The spheres were placed on a 25 micrometer screen to remove the
bath solution, after
which they were dried at about 40 C in the MiniGlatt fluidized bed dryer
(Glatt Air Techniques;
Ramsey, NJ) until the target moisture (about 5 to about 6%) was reached. The
particles were
sized to less than about 500 micrometers and greater than about 75
micrometers. The
composition, payload, and surface oil of the resultant product are illustrated
in the table below,
alpha- para- Linalyl Thymyl Can.ola Essential
Pinene Cylnene Linalool Acetate Th :mot , Acetate Oil Oil Total
wt % wt %. wt % wt % wt wt % wt % wt
wt %
Payload 1.96 7,5 N/A 2,6 NIA-7 14.6 7.0 267
33,7
Surface
oils 0,002 0,007 0.002 0,004 <0.002 0.063
0,58 0.076 0,654
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[00781 Example 6: Non-Esterified Functional Ingredient Mic.roencapsulated
in an
Alginate/Shellac Matrix
[0079] About 48 g of sodium alginate (ULV-L3G, Kimica Corp; Tokyo, Japan)
and about
g of sodium alginate (I-3G-150, Kimica Corp; Tokyo, Japan) were added to about
840 g of
water. Then, under agitated conditions, about 80 g of the pre-made shellac
solution w/ about
25% solids (Marcoat 125, Emerson Resources; Norristown., PA) was added. Next,
about 48 g of
the essential oil blend (about 24% soybean oil, about 4% alpha-pinene, about
30% para-cyinene,
about 7% linalool, and about 35% .thymol) was added and mixed and homogenized
until a fine,
stable emulsion was formed. Using a two-fluid nozzle, the solution was
atomized, creating
moderately small spheres of about 25to about 300 micrometers, into an aqueous
hardening bath
containing about 2.5% CaC12 and about 2.5% citric acid. After the particles
were cross-linked,
the particles were sieved on a 25 micrometer sieve and dried at about 40 C in
the MiniGlatt
fluid bed dryer (Glatt Air Techniques; Ramsey, NJ) until the target moisture
(about 5 to about
6%) was reached. Particles were sized at less than about 2/2 micrometers.
[00801 As seen in the table below, the payload was low and the particles
failed to mask the
undesirable taste/flavor of the essential oil when tasted in a model beverage
system. The
composition and payload of the resultant product are illustrated in the table
below.
alphatn para- Soybean Essential Oil Total
....... Pinene Cymene Limbo' Thyinol Oil .... Pa 'load Pa 'load
wt wt % wt % wt % wt % wt wt
Payload .01 I 2.8 .031 1.7 4.8 45 9.3
[0081] Example 7: Microencapsulation of an Essential Oil Blend Containing
Some
Esterified Components in an About 75% Alginate / About 25% Shellac Matrix
[0082] About 5,5 g of sodium alginate (1.11N-L3Gõ Kimica Corp; Tokyo,
Japan) and about
11 g of sodium alginate (1-3G-150, Kimica Corp; Tokyo, Japan) were added to
about 495 g of
water. Then, under agitation, about 22 g of the pre-made shellac (Temuss #594;
Ajax, Ontario,
Canada) in ammonium hydroxide solution (about 25% solids) was added. Next, the
essential
oil blend (about 18,5% canola oil, about 5.4% alpha-pinene, about 32.3% para-
cymene, about
.11.2% butyric acid ester of linalool (Malyl butyrate) and about 32.3% acetic
acid ester of thyrnol
(thymyl acetate)) was added and mixed and homogenized until a fine, stable
emulsion was
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formed with target droplet size of about 4 to about 7 micrometers, verified
with the Horiba
particle size analyzer (Horiba Industries; Irvine, CA), The solution was
atomized, creating
moderately small spheres between about 25 to about 300 micrometers, into an
aqueous
hardening bath containing about 2.5% CaCl2 and about 2.5% citric acid. The
particles were then
sieved on a 25 micrometer screen to remove the bath solution and then dried at
about 40C in the
MiniGlatt fluidized bed dryer (Glatt Air Techniques; Ramsey, NJ) until the
target moisture
(about 5 to about 6%) was reached. The particles were sized to less than about
212 micrometers.
100831 As shown in the table below and compared to Example 6, payload was
increased
significantly upon using a functional ingredient containing esterified
components. In parti-
cular, as can be seen in FIG. 2, an unexpected benefit of using an esterified
component is that
the payload of the non-esterified components also increased in the presence of
esterified
component. Further, the esterified components produced a less negative flavor!
taste impact
than the product formed in Example 6. The composition, payload, and surface
oil of the
resultant product are illustrated in the table below.
E
alpha- i para-
Unaly1 Thymyl Canola Essential
Pinene 00-Ilene Unatool Butyrate Thymol Acetate Oil
Oil
Total
wt 'f' Wt % wt % wt % wt % wt %
---,-- wt %
wt % wt %
Payload 1.80 9.2 N/A 3.9 N/A 10.8 8.0 25.70 33.70
_
¨
Surface
i
I oils 0.002 0.019 N/A 0.006 N/A . 0.066 ---------------- 0.61
0.093 ; 0.701
[00841 Example 8: Comparison of Payload Retention with Non-Esterified and
Esterified
Functional Ingredients within an About 75% Alginate / About25% Shellac Matrix
[00851 Comparison of the particles produced with the original essential oil
blend
(containing non-esterified components: alpha-pinene, para-cymene, linalool,
thymolõ and canola
oil) and particles produced with an essential oil blend containing some
esterified components
(alpha-pinene, para-cymene, linalyl butyrate, thymyl acetate, and canola oil)
is illustrated in
FIG. 2, The particles were produced. using .the same process conditions and
compositions aside
from the esterified versus non-esterified functional ingredients. In this
Figure, the level of
linalool (combined) is calculated by summing the measured linalool and the
linalool equivalent'
from the measured linaly1 acetate based on the molecular composition.
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[00861 As shown in FIG. 2, it is evident that the use of esterified
components in the essential
oil blend led to about a 130% increase in payload retention, the payload
retention increase seen
in both the esterified and non-esterified components of the functional
ingredient.
100871 Example 9: Effect of an Esterified Functional Ingredient on Gastric
Release
[0088] This example compares the release of the non-esterified functional
ingredient to the
esterified functional ingredient during a simulated gastrointestinal study
using an in vitro
digestion model.
[00891 FIG. 3 shows the known properties of the compounds. As shown in the
table of
FIG. 3, the solubility values for the ester compounds (linalyl acetate,
linalyl butyrate and thymyl
acetate) are significantly lower than the parent compounds (linalool and
thymol). In addition,
the partition coefficients for the ester compounds are greater than the parent
compounds.
These factors indicate that the ester compounds have a greater affinity for
the hydrophobic
carrier and insoluble matrix material than the parent compounds. FIGS. 4 and 5
show that the
release between -0.5 and 0.0 hours, representative of the residence time in
the simulated gastric
fluid, is greatly reduced for the particles containing some esterified
functional ingredient. The
reduced rate is most evident when comparing linalool and thymol release from
FIG. 4 to linalyl
acetate and thymyl acetate release from FIG. 5. This improvement in stability
within the model
stomach implies increased stability in other low pH systems, such as acidic
beverages, and
further illustrates the importance of the esters for successful
microencapsulation and enteric
release of the functional ingredients.
[00901 Example 10: Modulation of the Release from Microencapsulated
Particles and the
Hydrolysis of the Esterified Components
[00911 This example shows how the selection of various acids for
esterification can affect
the resulting rate of delivery of the parent compounds. Example 9 showed the
effect on release
rate from the particles within a gastric model, as a result of inclusion of
some esterified
compounds. As evident in FIGS. 4 and 5, the gastric release rate was reduced,
as well as the
release rate from 0-24.5 h, representative of the residence time in the small
intestine. FIGS. 6
and 7 show the release rates of two esters of linalool, linalyl acetate in
FIG. 6 and linalyl
butyrate in FIG. 7 within a digestion model simulating stomach and small
intestine conditions
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and residence times. In addition, the levels of the parent compounds present
in the digestion
model over time were measured. The presence of the parent compound, linalool
is a result of
hydrolysis of the linalyl acetate. As seen in FIG. 6, the initial release of
linalool was about 5%
and increases to about 20%, which correlates to about 33% hydrolysis of
linalyl acetate to
linalool. In FIG. 7, the initial release of linalool was about 2% and
increases to about 4%, which
correlated to about 5% hydrolysis of the linalyl butyrate to linalool. From
those results, it can be
seen that by formulating the functional ingredient with varying ratios of
linalyl acetate to linalyl
butyrate, the resulting level of linalool release through the gastric tract
and small intestine can
be modulated.
[00921 Example 11: Comparison of Model Beverage Systems with Esterified and
Non-Estezified Functional Ingredient
[0093] The taste profiles of two model beverages were compared. One
beverage was
prepared with particles comprising non-esterified functional ingredient with
the other beverage
was prepared with particles including esterified functional ingredients,
namely linalyl and
thymyl acetates. Particles were created in the same manner as Example 2. Two
equal,
one-serving amounts of powdered beverage mix were portioned out. To the first
portion, a
sufficient mass of the particles comprising the non-esterified functional
ingredient was added so
that about 70 mg of the functional ingredient was dosed. To the second
portion, a sufficient
mass of the particles including esterified functional ingredients were added
so that about 70 mg
of the functional ingredient was dosed. Each powder/particle mixture was then
added to about
200 ml of cold water and mixed thoroughly.
100941 Upon tasting samples of each model beverage, the panel concluded
that the model
beverage containing particles comprised of some esterified functional
ingredient had a
significantly reduced undesirable taste and/or flavor profile, leading to an
improved overall
sensory experience.
[0095] Example 12: Synthesis of a Dithymol Ester
[00961 In one case, a dithymol ester was produced by dissolving about 65
grams of thymol
and about 50 milliliters of pyridine in about 400 milliliters of hexane. With
the dissolved
combination being stirred at room temperature(about 20 to about 25 C), about
50 grams of
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sebacoyl chloride was added one drop at a time over a period of about 30 to
about 45 minutes.
After the sebacoyl chloride was added, the mixture was allowed to react
overnight at room
temperature (about 20 to about 25 C). The next day, the mixture was filtered
to remove solid
pyridine chloride. The clarified filtrate containing dithymol sebacate and
hexane was then
subjected to further purification by contact with solutions of about IN sodium
hydroxide, about
1N hydrochloric acid, and then water to remove any unwanted byproducts,
unreacted starting
materials, and residual pyridine. The purified dithymol sebacate in hexane was
then dried over
anhydrous sodium sulfate overnight to remove traces of water. The sodium
sulfate was
removed by filtration and the hexane removed by distillation to yield about 80
grams
(approximately 82% yield) of at least about 95 percent pure dithymol sebacate.
The identity and
purity of the final product was determined by gas-liquid chromatography and
mass
spectrometry.
[0097] Example 13: Preparation of Sodium Thymolate (Sodium Thymoxide)
[0098] In one case, sodium thymolate was produced by making a first
solution by
dissolving about 8 grams of sodium hydroxide in about 25 milliliters of water
followed by the
addition of about 225 milliliters of absolute ethanol. A second solution was
prepared by
dissolving about 31 grams of thymol in about 100 milliliters of absolute
ethanol. With the
second solution being stirred at room temperature (about 20 to about 25 C),
the first solution
was added one drop at a time to the second solution. The combined solution was
stirred
overnight. The next day, the combined solution was filtered to remove traces
of any
undissolved reactants or byproducts. The absolute ethanol, water, and residual
thymol were
then removed by placing the filtered combined solution under a vacuum
overnight at a
temperature of about 40 C. The dried sodium thymolate was then removed from
the flask to
yield about 32 grams (approximately 93% yield) of product. The dried sodium
thymolate could
be further purified to remove traces of residual thymol by admixing the sodium
thymolate with
hexane (about 2 milliliters of hexane per 1 gram of sodium thymolate),
filtering to remove the
hexane, and drying the further purified sodium thymolate under vacuum.
[0099] While the compositions and methods have been particularly described
with specific
reference to particular process and product embodiments, it will be
appreciated that various
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alterations, modifications, and adaptations may be based on the present
disclosure, and are
intended to be within the spirit of this disclosure.
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