Note: Descriptions are shown in the official language in which they were submitted.
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HERBAL SOLID FORMULATION AND PROCESS FOR PREPARING THE
SAME
Field of the Invention
This invention, in general relates to a herbal solid formulation. In
particular, the present
invention provides a herbal solid formulation comprising Andrographis
paniculata,
Terminalia arjuna, Azadirachta indica, Trikatu (Zingiber officinalis, Piper
longum, Piper
nigrum), Tinospora cordifolia or Ocimum sanctum extracts without using any
excipients and
preservatives and process for preparing the same.
Background of the Invention
Herbal supplements have been witnessed tremendous growth and acceptance among
the
consumers during the last decade due to their safety and efficacy. Unlike
allopathic
medications, herbal extracts are safe and devoid of any side effects. There is
a growing
concern among the consumers worldwide using naturally derived products and
avoiding
synthetic chemicals in their food, personal care products and daily health
supplements. Many
herbal products those are available in the market as tablets and capsule using
synthetic
excipients such as binders, lubricants and diluents and preservatives such as
Parabens and
salts of benzoic acids etc. These excipients and preservatives are reported to
have toxic and
side effects.
Pharmaceutical dosage forms such as tablets and capsules should have certain
properties such
as hardness, friability, disintegration time (DT), stability and delivery of
the drug to give
required therapeutic benefits to the patient. These properties are achieved
using the excipients
such as binders, lubricants and diluents.
It is therefore very important and challenging task to develop a process of
manufacturing
herbal tablets using herbal extract and plant powder without using any
synthetic excipient and
preservative.
Related Art
US Patent 6,207,189 by Mercati et al disclose a process for the production of
tablets and
capsules of natural substances of vegetable origin wherein dry extracts and
micronized
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powders of one or more medicinal herbs in appropriate proportions are blended
and subjected
to steam pressure followed by drying, preparation of granules and compression
to tablets.
US Patent 6,468,563 by Schmidt et al. discloses a process for producing
rapidly
disintegrating pharmaceutical formulation containing an extract and lubricant
and
compressing the blend to form the pharmaceutical formulation.
Summary of the Invention
It is a principal object of the invention to provide a herbal solid
formulation comprising
Andrographis paniculata, Terminalia arjuna, Azadirachta indica, Trikatu
(Zingiber
officinalis, Piper longum, Piper nigrum) Tinospora cordifolia or Ocimum
sanctum extracts
essentially free of additives/excipients and preservatives and providing
required quantity of
active constituents per dose.
It is another object of the invention to provide a herbal solid formulation of
herb
Andrographis paniculata, Terminalia arjuna, Azadirachta indica, Trikatu
(Zingiber
officinalis, Piper longum, Piper nigrum), Tinospora cordifolia or Ocimum
sanctum extracts
having reduced side effects and toxicity.
It is yet another object of the invention to provide a herbal solid
formulation of herb
Andrographis paniculata, Terminalia arjuna, Azadirachta indica, Trikatu
(Zingiber
officinalis, Piper longum, Piper nigrum) Tinospora cordifolia or Ocimum
sanctum extracts
essentially free of excipients/additives and preservatives and having desired
friability,
disintegration time and hardness.
It is still another object of the invention to provide a method for preparing
extract of herb
Andrographis paniculata, Terminalia arjuna, Azadirachta indica, Trikatu
(Zingiber
officinalis, Piper longum, Piper nigrum) Tinospora cordifolia or Ocimum
sanctum used to
prepare the solid formulation.
The above and other objects of the present invention are attained according to
following
preferred embodiments of the present invention. However the scope of the
invention is not
restricted to the particular embodiments discussed herein after.
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In accordance with one preferred embodiment of the present invention, there is
provided a
herbal solid formulation comprising a blend of Super Critical Fluid (CO2)
extract, water
extract and powder of herb Terminalia arjuna, Azadirachta indica, Trikatu
(Zingiber
officinalis, Piper longum, Piper nigrum) Tinospora cordifolia or Ocimum
sanctum, wherein
said blend of extract and said powder of herb is mixed in a ratio of about
1:0.5 to about 1:90.
In accordance with one preferred embodiment of the present invention, there is
provided a
herbal solid formulation comprises a blend of Super Critical Fluid (CO2)
extract, water
extract and powder of herb Terminalia arjuna, Azadirachta indica, Trikatu
(Zingiber
officinalis, Piper longum, Piper nigrum) Tinospora cordifolia or Ocimum
sanctum, wherein
said herbal solid formulation is essentially free of additives/excipients.
In accordance with another preferred embodiment of the invention there is
provided a process
for preparing a herbal solid formulation according to above essentially free
of
additives/excipients comprises of autoclaving powder of the herb, granulating
the autoclaved
powder with a water extract of the herb, lubricating the granulated mixture by
adding the
autoclaved powder of the herb and preparing the solid formulation.
In accordance with yet another preferred embodiment of the invention, the
powder of herb is
obtained by pulverizing the herb to a powder having mesh size preferably
between about 20
to about 100.
In accordance with yet another embodiment of the present invention, the
extract of the herb is
passed through a mesh having size between about 20 to 80.
In accordance with still another embodiment of the present invention, wherein
the granulation
of the blend of extracts and powder of the herb is carried out in presence of
a solvent,
preferably water and grain alcohol or combination thereof.
In accordance with yet another embodiment of the present invention there is
provided a
process for preparing the extract of the herb by Super Critical Fluid (CO2)
extraction,
percolation, hot soxhalation or refluxing followed by filtration and
concentration to dryness
at optimum temperature.
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In accordance with one other embodiment of the present invention, there is
provided a
process for sterilization of herbal powders by autoclaving the granular
mixture, wherein
autoclaving prevents microbial growth.
Brief Description of Drawings
Further objects of the present invention together with additional features
contributing thereto
and advantages accruing there from will be apparent from the following
description of
preferred embodiments of the invention which are shown in the accompanying
drawing
figures, wherein:
Figure 1 illustrates the LCMS chromatogram of Terminalia Arjuna supercritical
fluid CO2
extract.
Figure 2 illustrates the LCMS chromatogram of Terminalia Arjuna water extract.
Figure 3 illustrates LCMS chromatogram of Supercritical fluid CO2 extract of
Azadirachta
indica
Figure 4 illustrates LCMS chromatogram of water extract of Azadirachta indica
Figure 5 illustrates LCMS chromatogram of water extract of Trikatu
Figure 6 illustrates LCMS chromatogram of Supercritical fluid CO2 extract of
Trikatu
Figure 7 illustrates LC and total ion chromatogram of Guduchi (Tinospora) CO2
extract.
Figure 8 illustrates LC and total ion chromatogram of Guduchi (Tinospora)
water extract.
Figure 9 illustrates LC and total ion chromatogram of Tulsi (Ocimum) CO2
extract.
Figure 10 illustrates LC and total ion chromatogram of Tulsi (Ocimum) water
extract.
Detailed Description of the Invention
While this specification concludes with claims particularly pointing out and
distinctly
claiming that, which is regarded as the invention, it is anticipated that the
invention can be
more readily understood through reading the following detailed description of
the invention
and study of the included examples.
The present invention provides a herbal solid formulation essentially free of
excipients/additives or preservatives, wherein said formulation comprises a
blend of Super
Critical Fluid (C02) extract and water extract of Andrographis paniculata,
Terminalia
arjuna, Azadirachta indica, Trikatu (Zingiber officinalis, Piper longum, Piper
nigrum)
Tinospora cordifolia or Ocimum sanctum along with powder of Andrographis
paniculata,
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Terminalia arjuna, Azadirachta indica, Trikatu (Zingiber officinalis, Piper
longum, Piper
nigrum) Tinospora cordifolia or Ocimum sanctum and a process for preparing the
same.
The extracts of the herb is prepared by Super Critical Fluid (CO2) method.
Alternately the
same is prepared by employing percolation, hot soxhalation or refluxing method
using a
solvent, followed by filtration and concentration on a rotatory evaporator on
steam bath at
optimum temperature and under reduced pressure. The solvent employed includes
organic
grain alcohol, ethanol or water or combinations thereof, preferably grain
alcohol. The extract
is dried and passes through a mesh having size preferably between #20 to 80.
The powder of the herb is prepared by pulverizing the root of herb to a powder
of different
mesh sizes based on the requirement, preferably between about 20 to about 100,
more
preferably between 20 to 80. The extract and the powder of the herb is mixed
in a
predetermined ratio preferably between about 1: 0.5 to 1: 90 for optimum
granulation.
The process of preparing the herbal solid formulation involves granulation of
the blend of
extracts and powder of the herb using a solvent system, followed by passing
the granules
through a mesh having size preferably between 12 to 24 # and autoclaving the
granules. The
solvent system employed for granulating the mixture includes grain alcohol,
water or
combination thereof. Autoclaving helps in microbial control of the solid
formulation as it
does not contain any preservatives.
The autoclaved granules are further lubricated using the powder of the
Andrographis
paniculata, Terminalia arjuna, Azadirachta indica, Trikatu (Zingiber
officinalis, Piper
longum, Piper nigrum) Tinospora cordifolia or Ocimum sanctum and compressed or
encapsulated into tablets or capsules.
All extracts, granules and tablets are subjected to standardization by High
Performance Thin
Layer Chromatography (HPTLC) and High Performance Liquid Chromatography (HPLC)
for
identification and quantitative estimation of active marker compounds. The
extracts were
evaluated for toxicity studies in rats and tablets for safety studies in
healthy human
volunteers.
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The solid formulation according to the present invention has desired hardness
preferably
between 2 to 8 kg/cm2, more preferably about 3 to about 4 kg/cm2, friability
less than about
1% w/w and disintegration time less than about 60 min, preferably between 5 to
60 min, more
preferably less than about 30 min. The solid formulation complies with USFDA
guidelines.
According to the present invention, the disclosed solid formulation is
preferably granules,
tablet or capsule.
The following non-limiting examples illustrate specific embodiments of the
present
invention. They are, not intended to be limiting the scope of present
invention in any way.
Example-1
Preparation of Andrographis paniculata (Kalmeah) water extract
Approximately 100 Kg of shade dried plant material was subjected to extraction
with 400
Liters of purified water by percolation method at Room Temperature. The water
extractions
after 24 hours were filtered through muslin cloth and concentrated to thick
paste. After
achieving the desired total solid content, the soft extract was spray dried to
a free flowing dry
extract powder.
Example-2
Preparation of Andrographis paniculata (Kalmegh) SCFE (CO2) Extract
Approximately 25 Kg of The whole plant of Andrographis paniculata was
pulverized to fine
powder and loaded in to extractor. Super Critical Carbon dioxide liquid was
pmped in to the
extractor at a pressure of 300 bar and 39 C temperature for 2-3 hours. Extract
was separated
into the container at pressure of 40 bar and 20 C temperature. The CO2 super
critical liquid
was recycled from the extraction vessel. The resultant extract was analyzed
for active
markers of Andrographolides.
Example-3
Formula of Granulation
Table 1
S. Name of the material Weight per Weight per
No. Tablet (mg) Batch (Kg)
I Andrographispaniculata (Water extract) (# 40 mesh) 200.00 20.00
2 Andrographis paniculata (Super critical fluid (CO2) extract) 50.00 5.00
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L 3 Andrographis paniculata (#40) powder 375.00 37.50
Example-4
Formula of Lubrication of granules
Table 2
S. Name of the material Weight per Weight per
No. Tablet (mg) Batch (Kg)
1 Andrographis paniculata Extract Granules (# 16 mesh) 625.00 62.50
2 Andrographispaniculata (#40) powder 25.00 2.50
Total 650.00 65.00
Example-5
Manufacturing procedure of granulation and compression of Tablet
1. Dispense Extract, Plant powder as per Batch record.
2. Transfer Andrographis paniculata Leaf powder (#40 mesh) to stainless steel
trays and
sterilize at 160 C for 120 minutes.
3. Check the sterilized material for LOD (Loss on Drying), Bulk Density and
Microbial
growth.
4. Sift 25 Kg of Andrographis paniculata Water extract through #40 mesh and
37.50 Kg
and 2.50 Kg of Leaf powder through # 40 mesh. Keep the materials packed in
double
poly bags.
5. Charge 25 Kg of Water extract and 37.50 Kg of Leaf powder in to RMG and mix
slowly for 5 minutes. Add 5 Kg of Andrographis paniculata CO2 extract and 2.5
Kg
of Purified water to the mixture over a period of 3 minutes with medium speed.
6. Stop the mixer and scrap off the material from the sides and bottom and
continue
mixing by operating impeller mixing at high speed with chopper ON for 3
minute.
7. Discharge the material from RMG
8. Mill the wet mass obtained from the above procedure in Multi mill fitted
with 8 mm
screen.
9. Dry the wet mass obtained through Multi mill at 70 C to 80 C for 60 minutes
and
checks the moisture every 30 minutes to achieve LOD at 2.5 to 3.5%.
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10. Sift the dried granules through #16 SS mesh and store in double polylines
HDPE
containers.
11. Analyze granules for LOD, BD, micro analysis, granules-fine ratio etc.
12. Maintain the temperature at 25 C and relative humidity NMT 40% of blending
and
compression area.
13. Transfer the sifted Leaf powder to blending area and blend the leaf powder
and
granules in to the double cone blender for 3 minutes at 20-25 RPM.
14. Compress the approved granules into tablets with punch size 17 X 8 mm size
with
upper and bottom as plain surface.
15. Check appearance, thickness and hardness of tablets every 30 minutes.
16. Check friability and DT for every 1 hour and average weight every 30
minutes.
17. Collect the tablets in double poly lined air tight container.
18. Final tablets have the following specifications:
STANDARD PARAMETERS *
1. Theoretical average weight: 650 mg
2. Weight uniformity : 650 mg 5% (617.5mg to 682.5mg)
3. Weight of 20 tablets : 13.00 g 3% (12.61gm to 13.39gm)
4. Tablet thickness : 4.8 to 5.8 mm
5. Tablet hardness : 4 to 6 Kg/cm2
6. Friability : NMT 1.0% W/W
7. Disintegration time : NMT 30 min.
8. Andrographolide : 9 mg per caplet
Example-6
Preparation of Terminalia arjuna (Arjuna) water extract
Approximately 100 Kg of shade dried plant material bark of Terminalia arjuna
was subjected
to extraction with 400 Liters of purified water by percolation method at Room
Temperature.
The water extractions after 24 hours were filtered through muslin cloth and
concentrated to
thick paste. After achieving the desired total solid content, the soft extract
was spray dried to
a free flowing dry extract powder. The residual plant materials were named as
spent powder.
Example-7
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Preparation of Terminalia arjuna (Arjuna) SCFE (CO,) Extract
Approximately 25 Kg of bark of Terminalia arjuna was pulverized to fine powder
and loaded
in to extractor. Super Critical Carbon dioxide liquid was pumped in to the
extractor at a
pressure of 300 bar and 39 'C temperature for 2-3 hours. Extract was separated
into the
container at pressure of 40 bar and 20 'C temperature. The CO2 super critical
liquid was
recycled from the extraction vessel.
Example-8
Formula of Granulation (Table-3)
S. Name of the material Weight per Weight per
No. Tablet in mg Batch in Kg
1 Terminalia arjuna (Water extract) (# 40 mesh) 240.00 24.00
2 Terminalia arjuna (Super critical fluid (CO2) extract) 10.00 1.00
3 Terminalia arjuna ( spent powder) 200.00 20.00
4 Terminalia arjuna leaves (#40) powder 150.00 15.00
Purified water Granulation Quantity
fluid sufficient
Example-9
Formula of Lubrication of granules (Table-4 )
S. Name of the material Weight per Weight per
No. Tablet in mg Batch in Kg
1 Terminalia arjuna Extract Granules (# 16 mesh) 600.00 60.00
2 Terminalia arjuna spent powder (#40) powder 100.00 10.00
Total 700.00 70.00
Example-10
DISPENSING OF RAW MATERIAL
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1. Dispense the raw materials as per Batch Formula
DRY HEAT STERILIZATION
1. Transfer 30.00 kg of spent powder of Terminalia arjuna bark (left material
after
water extraction) and 15 kg of leaves of Terminalia arjuna into trays and
sterilize the
material @ 160'C for 60 mins.
2. Unload the sterilized materials in to double lined polybags separately and
keep in
airtight containers.
3. Send the sample for LOD, BD and Microbial Analysis. Table-5.
Table-5
Parameter Standard Value
1 LOD
spent powder of Terminalia arjuna bark 2.0-3.0%w/w
leaves powder of Terminalia arjuna 2.0-2.30% w/w
2 BD
spent powder of Terminalia arjuna bark 0.3-0.5 g/ml
Leaves powder of Terminalia arjuna 0.2-0.5 g/ml
3 TVAC NMT 5000 cfu/gm
4 Fungal count NMT 10 cfu/gm
SIFTING
1. Sift 24.00 kg of water extract of bark of Terminalia arjuna through # 40.
2. Sift 20.00 kg of spent powder of Terminalia arjuna bark and 15.00 kg of
Leaves
powder of Terminalia arjuna through #60.
3. Sift 6.30 kg of spent powder of Terminalia arjuna bark lubricant through #
40
separately.
4. Collect the above-sifted materials in separate duly labeled double lined
polybags.
5. Record Quantity Sifted and the sieve integrity before and after sifting.
PREPARATION OF GRANULATION FLUID
1. Transfer about 5.0 kg of purified water into a clean Stainless Steel
Vessel.
2. Record the observations. Record deviations if any.
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GRANULATION
1. Charge 15.00 Kg of leaves powder of Terminala arjuna and then add 1.00 kg
of super
critical (C02) extract of Terminalia arjuna for about 5 min.
2. Charge 20.00 kg of spent powder of Terminalia arjuna bark into RMG and
blend for
about 5 min.
3. Charge 24.00 kg of water extract of bark of Terminalia arjuna into RMG and
blend
for about 5 min.
4. Slowly add the granulation fluid to the RMG over a period of about 3 min,
with
medium speed.
5. Stop the mixer and scrape off the mass from the sides and bottom.
6. Continue mixing by operating the impeller at high speed with Chopper ON for
about
3 minute.
7. Add additional quantity of purified water, if required.
8. Discharge the mass from the RMG.
9. Record the observations. Record deviations if any.
WET MILLING
1. Mill the Wet Mass in Multi mill fitted with 8 mm screen.
TRAY DRYING
1. Dry the Wet mass in tray Drier at about 70 C to 80 C for about 60 minutes.
2. Check the Moisture once every 30 minutes. (LOD 4.0 to 6.0% w/w) and record
the
details.
SIZING
1. Sift the dried granules using a Sifter fitted with sieve #16.
2. Collect the sifted granules in a clean double poly lined HDPE container.
3. Mill the retains (oversize granules) through a Multi Mill fitted with 1.5mm
screen
with `Knives Forward' direction.
4. Pass the milled granules through a Sifter fitted with # 16 sieve.
5. Collect the sized granules and add them to the sifted granules.
6. Weigh the granules. Record deviations if any.
7. Analyse samples as per below table-6.
8. If required autoclave the granules at 120 C for 20 min.
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Table-6
Parameter Standard values
I LOD 4.0-6.0%w/w
2 BD 0.40 - 0.60 g/ml
3 Actives As per finished product spec
4 TVAC NMT 5000 cfu/gm
Fungal count NMT 10 cfu/gm
BLENDING
Maintain the Temperature and Relative Humidity of the area with in the
specified limit (Limit
Temperature NMT 25 -C and relative humidity NMT 40%)
1. Transfer the sized granules to the blending area.
2. Transfer the sifted spent powder of Terminalia arjuna lubricant to the
blending area.
3. Load 10.00 kg of spent powder of Terminalia arjuna lubricant and the sized
granules
into the Double Cone blender.
4. Blend the ingredients for 3 minutes at 20-25 RPM.
5. Record the details.
6. Unload the blend in a clean Double poly-lined HDPE drums and affix duly
filled
status labels.
7. Weigh the blend and enter the details.
COMPRESSION
Maintain the Temperature and Relative Humidity of the area with in the
specified limit
(Limit Temperature NMT 25 *C and relative humidity NMT 40%)
1. Check the Temperature and Relative Humidity of the area and record.
2. Bring the drums containing the blend into the compression area.
3. Adjust the machine.
4. Carry out the initial checks before starting the operation as specified in
Table -7.
Table-7
Description
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Punch Size 17x 8 mm caplet
Upper Punch Plain
Lower Punch Plain
5. Check for Appearance, Average Weight, Individual weight, Thickness,
Hardness,
Friability & Disintegration of 6 tablets.
6. Check appearance, thickness & hardness of tablets every 30 min.
7. Check Friability and DT every 1-hour.
8. Check Average weight of 20 tablets every 30 mins graphically.
9. Collect the tablets in a double poly-lined, tightly closed, container.
Weigh each
container and enter the details.
STANDARD PARAMETERS Of TABLETS
Standard parameters are mentioned in Table-8
Table-8
SI.No. Parameters Standard value
1 Theoretical average weight 700 mg
2 Weight uniformity 700 mg 5% (665 mg to 735 mg)
3 Weight of 20 tablets 14.00 g :L 3%
4 Tablet thickness 4.5 to 5.5 mm
Tablet hardness 3 to 5 Kg/cm
6 Friability NMT 1.0% W/W
7 Disintegration time NMT 30 min.
8 Arjunolic acid 0.25 mg
Tannins 120 mg
1. Collect the Compressed tablets in Double poly lined HDPE drums and record
their
weights.
2. Affix duly filled status labels to the containers.
PACKING
1. Pack 60 tablets in a HDPE 120 CC container and weigh them for
appropriateness of
number of tablets.
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Example-11
Estimation of arjunolic acid in Arjuna dry extract, granules
Standard arjunolic acid solution (0.1 mg/ml): Weigh accurately 10 mg of
standard arjunolic
acid in a 10 ml volumetric flask. Add 7 - 8 ml of methanol and dissolve. Make
the volume up
to the mark with methanol. Take 1 ml of this solution in another 10 ml
volumetric flask,
dilute and make the volume up to the mark with methanol.
Sample preparation (10 mg/ml): Weigh accurately 0.5 g of sample in a 250 ml
flat bottomed
flask. Add 50 ml of methanol and reflux it on a water bath at 80 'C for 30
minutes. Filter and
transfer it into a 50 ml volumetric flask. Make the volume up to the mark with
methanol.
HPLC Conditions were as follows
Column: C18 ODS Hypersil (250 x 4.6) particle size: 5 p (Make: Thermo)
Mobile phase: Methanol: 0.1% Phosphoric acid (70: 30)
Flow rate: 1 ml/minute
Wavelength: 210 nm
Chromatographic procedure: Stabilize the instrument with the above mentioned
mobile
phase. Inject 20 l of standard solution and record the chromatogram.
Similarly, inject 20 l
of sample solution and record the chromatogram. Calculate the area of standard
peak and the
corresponding peak in the sample.
Calculation: % w/w Arjunolic acid content can be calculated using the formula.
Area of sample Concentration of standard
w/w peak (mg/ml)
------------------------ ------------------------------------- % Purity of
Arjunol = -- ----- x ---------- x
standard
is acid
Area of standard Concentration of sample
peak (mg/ml)
Example-12
LCMSMS STUDIES
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Liquid Chromatography -Mass Spectrometer Analysis of Terminalia arjuna bark
supercritical (C02) extract and water extract (Figure 1 and 2):
LCMSMS analysis were carried out by using an applied biosystem-Sciex API 2000
triple
quadrupole mass spectrometer equipped with an ion source turbo spray and ESI
interface.
The liquid chromatography was a LC-20 AD series binary system equipped with an
autosampler. The column used was C18 phenomenex (250 X 4.6 mm, 5 m), flow rate
0.5
mi/min of mobile phase methanol: water (0.1% acetic acid), (10:90), wave
length 275 nm and
run time 25 min. The analytes were ionized by ESI in positive-ion mode (PI
mode). Final
ionization conditions were heated ionization temperature 435 C. Curtain gas
Nitrogen 30 psi,
particulate -free and CO2- free air was used as ion source gas I at a flow
rate 50 psi and ion
source gas 2 at a flow rate 60 psi.
Sample preparation for Terminalia arjuna bark CO2 extract:
Weigh about 50 mg of sample in a 50 ml volumetric flask, and dissolve by
sonication in
methanol (HPLC grade). Make the volume up to the mark with methanol filter
through 0.2
urn syringe filter.
Sample preparation for Terminalia arjuna bark water extract:
Weigh about 500 mg of finely powdered sample in a 50 ml volumetric flask, and
dissolve by
sonication in water (HPLC grade). Make the volume up to the mark with water
filter through
0.2 urn syringe filter.
Example-13
Preparation of Azadirachta indica (Neem) water extract
Approximately 100 Kg of shade dried plant material leaves of Azadirachta
indica was
subjected to extraction with 400 Liters of purified water by percolation
method at Room
Temperature. The water extractions after 24 hours were filtered through muslin
cloth and
concentrated to thick paste. After achieving the desired total solid content,
the soft extract
was spray dried to a free flowing dry extract powder.
Example-14
Preparation of Azadirachta indica (Neem) SCFE (CO2) Extract
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Approximately 25 Kg of leaves of Azadirachta indica was pulverized to fine
powder and
loaded in to extractor. Super Critical Carbon dioxide liquid was pumped in to
the extractor at
a pressure of 300 bar and 39'C temperature for 2-3 hours. Extract was
separated into the
container at pressure of 40 bar and 20'C temperature. The CO2 super critical
liquid was
recycled from the extraction vessel.
Example- 15
Formula of Granulation (Table-9)
S. Name of the material Weight per Weight per
No Tablet in mg Batch in Kg
1 Azadirachta indica leaves powder (Water extract) (# 40) 200.00 20.00
Azadirachta indica leaves (Super critical fluid
2 80.00 8.00
(C02)extract)
3 Azadirachta indica stem (#100) powder 300.00 30.00
4 Granulatio Quantity
Purified water
n fluid sufficient
Example-16
Formula of Lubrication of granules (Table-10)
S. Name of the material Weight per Weight per
No. Tablet in mg Batch in Kg
1 Azadirachta indica Extract Granules (# 16 mesh) 580.00 58.00
2 Azadirachta indica stem powder (#100) powder 20.00 02.00
Total 600.00 60.00
Example-17
Manufacturing Details Of Tablets
Dispensing of Raw Material
1. Dispense the raw materials as per Batch record.
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Dry Heat Sterilization
1. Transfer 30 kg and 2 kg of stem powder of Azadirachta indica into trays and
sterilize the material at 160 C for 60 mins.
2. Unload the materials in to double lined polybags separately and keep in
airtight
containers.
3. Analyse the sample for LOD, BD and Microbiological Analysis as per Table-11
Table-11
Parameter Standard values
1 LOD 2.0-4.0%w/w
2 BD 0.20-0.30 g/ml
3 TVAC NMT 5000 cfu/gm
4 Fungal count NMT 10 cfu/gm
Siftin
1. Sift 20.00 kg of water extract of leaves of Azadirachta indica through # 40
2. Sift 08.00 kg of Coe extract of leaves Azadirachta indica through # 40
3. Sift 30.00 kg of stem powder of Azadirachta indica through # 100
4. Sift 2.00 kg of stem powder of Azadirachta indica through # 100 separately
5. Collect the above-sifted materials in separate duly labeled double lined
polybags.
6. Record the Quantity Sifted and the sieve integrity before and after sifting
Preparation of granulation fluid
1. Transfer about 20.0 kg of Purified water into a clean Stainless Steel
Vessel
Granulation
1. Charge 20.00 Kg of water extract of leaves of Azadirachta indica, 8.0 kg of
CO2
extract of leaves Azadirachta indica and 30.00 kg of stem powder of
Azadirachta
indica into the RMG, mix for about 5 minute.
2. Slowly add the granulation fluid to the RMG containing the Sifted materials
over
a period of about 3 minute, with medium speed.
3. Stop the mixer and scrape off the mass from the sides and bottom.
4. Continue mixing by operating the impeller at high speed with Chopper ON for
about 3 minute.
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5. Add additional quantity of Purified water, if required.
6. Discharge the mass from the RMG.
7. Record the observations. Record deviations if any.
Wet Milling
1. Mill the Wet Mass in Multi mill fitted with 8mm screen.
Tray Drying
1. Dry the Wet mass in tray drier at about 70 C to 80 C for about 60
minutes.
2. Check the Moisture once every 30 minutes. (LOD Limit: 3 to 4% w/w) and
record
the details
Sizing
1. Sift the dried granules using a Sifter fitted with sieve #16.
2. Collect the sifted granules in a clean double polylined HDPE container.
3. Mill the retains (oversize granules) through a Multi Mill fitted with 1.5
mm screen
with `Knives Forward' direction
4. Pass the milled granules through a Sifter fitted with #16 sieve.
5. Collect the sized granules and add them to the sifted granules
6. Weigh the granules.
7. Analyse the sample as per Table-12
Table-12
Parameter Standard values
I LOD 2.5-3.5 %w/w
2 BD 0.30 - 0.40 g/ml
3 Granules to fine ration 70:30
4 TVAC NMT 5000 cfu/gm
Fungal count NMT 10 cfu/gm
Blending
1. Transfer the sized granules into the blending area.
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2. Transfer the sifted stem powder of Azadirachta indica lubricant into the
blending
area.
3. Load 5 kg of stem powder of Azadirachta indica and the sized granules into
the
Double Cone blender.
4. Blend the ingredients for about 3 minutes at 20-25 RPM.
5. Record the details
6. Unload the blend in a clean Double poly-lined HDPE drums and affix duly
filled
status labels.
7. Weigh the blend and enter the details.
Compression
Maintain the Temperature and Relative Humidity of the area with in the
specified limit (Limit
Temperature NMT 25'C and relative humidity NMT 40%)
1. Check the Temperature and Relative Humidity of the area and record.
2. Bring the drums containing the blend into the compression area.
3. Adjust the machine as per the parameters.
4. Carry out the initial checks before starting the operation as specified in
below
Table-13
Table- 13
Description
Punch Size 17x 8 mm caplet
Upper Punch Plain
Lower Punch Plain
5. Check for Appearance, Average weight, Individual weight, Thickness,
Hardness,
Friability & DT of 6 tablets.
6. Check appearance, thickness & hardness of tablets every 30 min.
7. Check Friability and DT every 1-hour.
8. Check Average weight of 20 tablets every 30 mins graphically.
9. Collect the tablets in a double poly-lined, tightly closed, container.
Weigh each
container and enter the details.
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STANDARD PARAMETERS OF TABLETS
Standard parameters are mentioned in Table- 14
Table-14
S. No. Parameters Standard value
I Theoretical average weight 600 mg
2 Weight uniformity 600 mg 5% (570mg to
630mg)
3 Weight of 20 tablets 12.0 g 3% (11.64gm to
12.36gm)
4 Tablet thickness 4.0 to 5.0 mm
Tablet hardness 4 to 6 Kg/cm2
6 Friability NMT 1.0% W/W
7 Disintegration time NMT 30 min.
8. Bitters containing Nimbidin 18 mg per caplet
10. Collect the Compressed tablets in Double polylined HDPE drums and record
their
weights.
11. Affix duly filled status labels to the containers.
Packing
1. Pack 60 tablets in a HDPE 120 CC container and weigh them for
appropriateness
of number of tablets.
Example-18
LCMSMS STUDIES
Liquid Chromatography -Mass Spectrometer Analysis of Azadirachta indica
leaves,
supercritical (CO2) extract and water extract (Figure 3 and 4):
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LCMSMS analysis were carried out by using an applied biosystem-Sciex API 2000
triple
quadrupole mass spectrometer equipped with an ion source turbo spray and ESI
interface.
The liquid chromatography was a LC-20 AD series binary system equipped with an
autosampler. The column used was C18 phenomenex (250 X 4.6mm, 5 m), flow rate
0.5mllmin of mobile phase methanol: water (0.1% acetic acid), (10:90), wave
length 254 nm
and run time 25 min. The analytes were ionized by ESI in positive-ion mode (PI
mode). Final
ionization conditions were heated ionization temperature 435 C. Curtain gas
Nitrogen 30 psi,
particulate -free and C02- free air was used as ion source gas I at a flow
rate 50 psi and ion
source gas 2 at a flow rate 60 psi.
Sample preparation for Azadirachta indica leaves CO2 extract:
Weigh about 50mg of sample in a 5 Oml volumetric flask, and dissolve by
sonication in
methanol (HPLC grade). Make the volume up to the mark with methanol filter
through 0.2
um syringe filter.
Sample preparation for Azadirachta indica leaves water extract:
Weigh about 500 mg of finely powdered sample in a 50 ml volumetric flask, and
dissolve by
sonication in water (HPLC grade). Make the volume up to the mark with water
filter through
0.2 um syringe filter
Example-19
Preparation of Trikatu (Zingiber ocinale rhizome, Piper longum fruits, Piper
nigrum fruits,
(1:1:1)) water extract
Approximately 100 Kg of shade dried plant material was subjected to extraction
with 450
Liters of purified water by percolation method at Room Temperature. The water
extractions
after 24 - 48 hours were filtered through muslin cloth and concentrated to
thick paste. After
achieving the desired total solid content, the soft extract was spray dried to
a free flowing dry
extract powder. The water extract was also prepared by hot soxhlation method.
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Example-20
Preparation of Trikatu (Zingiber officinale rhizome, Piper longum fruits,
Piper nigrum fruits,
(1:1:1)) SCFE (COQ Extract
Approximately 25 Kg of Trikatu material was pulverized to fine powder and
loaded in to
extractor. Super Critical Carbon dioxide liquid was pumped in to the extractor
at a pressure of
325 bar and 32 'C temperature for 2-4 hours. Extract was separated into the
container at
pressure of 45 bar and 21'C temperature. The CO2 super critical liquid was
recycled from the
extraction vessel.
Example-21
Batch Formula for Preparation of Organic Trikatu Granules (Formula- 1) , Table-
15
Table-15
S. Raw Material Weight per Weight per
No. Tablet in mg Batch In kg
1 Water extract of Trikatu (60# mesh) 280.00 28.00
Super critical fluid extract (SCFE, CO2 extract)
2 20.00 2.00
of Trikatu
4 Trikatu plant powder (60 # mesh) 340.00 34.00
Purified Water Qs * Qs
Formula of Lubrication of Trikatu granules, Table-16
Table-16
1 Trikatu granules (#16mesh) Granules 640.00 64.00
2 Trikatu plant powder (60# mesh) Lubricant 50.00 5.00
Total 690.00 69.00
Example-22
Manufacturing details of Trikatu tablets
DISPENSING OF RAW MATERIAL
1. Dispense the raw materials as per Batch record.
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DRY HEAT STERILIZATION
1. Transfer Trikatu plant powder into trays and sterilize the material at 160
C for 2
hr.
2. Unload the sterilized material in to double lined poly bags separately and
keep in
airtight containers.
3. Analyse the sample for LOD, BD and Microbiological Analysis. Table- 17
Table-17
Parameter Standard values
1 LOD 1.0-4.0%w/w
2 BD 0.25 - 0.60 g/mI
3 TVAC NMT 5000 cfu/gm
4 Fungal count NMT 10 cfu/gm
SIFTING
1. Sift 5.0 kg of water extract powder of Trikatu through # 60.
2. Sift 34.0 and 5.0 kg of Trikatu plant powder through # 60.
3. Collect the above-sifted materials in separate duly labeled double lined
poly bags.
PREPARATION OF GRANULATION FLUID
1. Transfer about 10 kg of Purified water into a clean Stainless Steel Vessel.
GRANULATION
I. Charge water extract powder of Trikatu, and Trikatu plant powder into the
RMG,
mix for about 5 minutes.
2. Slowly add super critical fluid extract (C02) of Trikatu, and granulation
fluid to
the RMG containing the Sifted extract powder of Trikatu, and Trikatu plant
powder over a period of about 3 minute, with medium speed.
3. Stop the mixer and scrape off the mass from the sides and bottom.
4. Continue mixing by operating the impeller at high speed with Chopper ON for
about 3 minute.
5. Add additional quantity of granulation fluid, if required.
6. Discharge the mass from the RMG.
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WET MILLING
1. Mill the Wet Mass in Multi mill fitted with 8mm screen.
TRAY DRYING
1. Dry the Wet mass obtained in tray Drier at about 60 C to 70 C for about
60
minutes.
SIZING
1. Sift the dried granules from Sifter fitted with sieve #16.
2. Collect the sifted granules in a clean double poly lined HDPE container.
3. Mill the retains (oversize granules) obtained through a Multi Mill fitted
with mm
screen with `Knives Forward' direction
4. Pass the milled granules through a Sifter fitted with 416 sieve.
5. Weigh the granules analyse as per Table-18
Table- 18
Parameter Standard values
1 LOD 2.0- 5.0 % w/w
2 BD 0.30 - 0.60g/ml
3 TVAC NMT 5000 cfu/gm
4 Fungal count NMT 10 cfu/gm
BLENDING
1. Transfer the sized granules into the blending area.
2. Transfer the sifted trikatu herb powder lubricant into the blending area.
3. Load trikatu herb powder and the sized granules into the Double Cone
blender.
4. Blend the ingredients for 6 minutes at 10-11 RPM.
5. Record the details
6. Unload the blend in a clean Double poly-lined HDPE drums and affix duly
filled
status labels.
7. Weigh the blend and enter the details.
ay
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COMPRESSION
1. Check the Temperature and Relative Humidity of the area and record.
2. Bring the drums containing the blend into the compression area.
3. Adjust the machine.
4. Carry out the initial checks before starting the operation as specified in
below.
Table-19
Description
Punch Size 17x8 mm caplet
Upper Punch Plain
Lower Punch Plain
5. Check for Appearance, Average Weight, Individual weight, Thickness,
Hardness,
Friability & DT of 6 tablets.
6. Check appearance, thickness & hardness of tablets every 30 min.
7. Check Friability and DT every 1-hour.
8. Check Average weight of 20 tablets every 30 mins graphically.
9. Collect the tablets in a double poly-lined, tightly closed, container.
Weigh each
container and enter the details.
Example-23
Finished product specification of Trikatu per caplet. Table-20
Table-20
Parameters Standard Range
Theoretical Average weight 690 mg
Weight uniformity 690 mg 5% (655 mg to 725 mg)
Weight of 20 tablets 13.8 g 3% (13.39 gm to 14.21 gm
Tablet thickness 3.5 to 6.5 mm
Tablet hardness 2 to 8 Kg/cm2
Friability NMT 1.0% W/W
Disintegration time NMT 30 min
Gingerols 3 mg per caplet
Piperine 1.09 mg per caplet
1. Tablets should be packed immediately after compression.
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2. The granules can also be filled in capsules.
PACKING
1. Pack 60 tablets in a HDPE 120 CC container and weigh them for
appropriateness of
number of tablets.
Example-24
Liquid Chromatography -Mass Spectrometer Analysis of Trikatu water extract:
(Figure 5)
LCMSMS analysis was carried out by using an applied biosystem-Sciex API 2000
triple
quadrupole mass spectrometer equipped with an atmospheric pressure chemical
ionization
source and heated nebulizer APCI interface. The liquid chromatography was a LC-
20 AD
series binary system equipped with an autosampler. The column used was C18
phenomenex
(250 x 4.6 mm, 5 m), flow rate 1 ml/min of mobile phase water (0.1% acetic
acid) (100%),
wave length 275 nm and run time 20 min. The analytes were ionized by APCI in
positive -
ion mode (PI mode). Final ionization conditions were heated nebulizer
temperature 450 ' C,
curtain gas Nitrogen 25 psi, particulate - free and CO2 - free air was used as
nebulising gas
at a flow rate of 70 psi.
Sample preparation for Trikatu water extract:
Weigh about 500mg of sample in 50ml volumetric flask, and dissolve by
sonication in water.
Make the volume up to the mark with water and filter through 0.2 m syringe
filter.
LCMSMS chromatograms are presented in Figure-5
Liquid Chromatography -Mass Spectrometer Analysis of Trikatu Super Critical
Extract
(CO,) extract: (Figure 6)
LCMSMS analysis were carried out by using an applied biosystem-Sciex API 2000
triple
quadrupole mass spectrometer equipped with an atmospheric pressure chemical
ionization
source and heated nebulizer APCI interface. The liquid chromatography was a LC-
20 AD
series binary system equipped with an autosampler. The column used was C 18
phenomenex
(250 X 4.6mm, 5 m), flow rate Iml/min of mobile phase methanol: water (0.1%
acetic acid)
(65 : 35), wave length 275nm and run time 20 min. The analytes were ionized by
APCI in
positive -ion mode (PI mode). Final ionization conditions were heated
nebulizer temperature
450 C, curtain gas Nitrogen 25 psi, particulate - free and CO2 - free air was
used as
nebulising gas at a flow rate of 70 psi.
2 (o
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Sample preparation for Trikatu super critical extract (C02) extract:
Weigh about 50 mg of sample in 50 ml volumetric flask, and dissolved by
sonication in
methanol. Make the volume up to the mark with methanol and filter through 0.2
m syringe
filter.
LCMSMS chromatogram are presented in Figure-6
Example-25
Preparation of Tinospora cord folia (Guduchi) water extract
Approximately 100 Kg of shade dried plant material stem was subjected to
extraction with
400 Liters of purified water by percolation method at Room Temperature. The
water
extractions after 24 hours were filtered through muslin cloth and concentrated
to thick paste.
After achieving the desired total solid content, the soft extract was spray
dried to a free
flowing dry extract powder.
Example-26
Preparation of Tinospora cordifolia Guduchi) SCFE (C0? Extract
Approximately 25 Kg of the stem of Tinospora cordifolia was pulverized to fine
powder and
loaded in to extractor. Super Critical Carbon dioxide liquid was pumped into
the extractor at
a pressure of 300 bar and 39 OC temperature for 2-3 hours. Extract was
separated into the
container at pressure of 40 bar and 20 'C temperature. The CO2 super critical
liquid was
recycled from the extraction vessel.
Example-27
Formula of Granulation (Table-21 )
S. Name of the material Weight per Weight per
No. Tablet in mg Batch in Kg
1 Tinospora cordifolia stem (Water extract) (# 40 mesh) 200.00 20.00
2 Tinospora cordifolia stem (Super critical fluid (CO2) 50.00 5.00
extract)
3 Tinospora cordifolia stem (#40 mesh) powder 400.00 40.00
Granulation Quantity
4 Purified water
fluid sufficient
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Example-28
Formula of Lubrication of granules (Table-22)
S. Name of the material Weight per Weight per
No. Tablet in mg Batch in Kg
1 Tinospora cordifolia Extract Granules (# 16 mesh) 650.00 65.00
2 Tinospora cordifolia stm powder (#80) powder 50.00 5.00
Total 700.00 70.00
Example-29
DISPENSING OF RAW MATERIAL
1. Dispense the raw materials as per Batch Formula.
DRY HEAT STERILIZATION
1. Transfer 45.0 kg of stem powder of Tinospora cordifolia in trays and
sterilize the
materials at 160 'C for 2 hour.
2. Unload the sterilized materials in to double- lined poly bags separately
and keep in air
tight containers.
3. Analyse the sample for LOD, BD and microbial analysis. Table-23
Table-23
Parameter Standard values
I LOD 1.0-4.0%
2 BD 0.25 - 0.60 g/ml
3 TVAC NMT 5000 cfu/gm
4 Fungal Count NMT 10 cfu/gm
SIFTING
1. Sift 20.0 kg of water extract of stem of Tinospora cordifolia through # 40.
2. Sift 5.0 kg of water extract of stem of Tinospora cordifolia through # 40.
3. Sift 40.0 kg of stem plant powder through # 80.
4. Sift 5.00 kg of stem plant powder lubricant through # 80 separately.
5.Collect the above-sifted materials in separate duly labeled double lined
polybags.
2g
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6.Record Quantity Sifted and the sieve integrity before and after sifting.
PREPARATION OF GRANULATION FLUID
1. Transfer about 20.0 kg of Purified water into a clean Stainless Steel
Vessel
GRANULATION
1. Charge 20.00 Kg of water extract of stem of Tinospora cordifolia, 5.0 kg of
super
critical extract (C02) of stem of Tinospora cordifolia, 40.0 kg of stem plant
powder into the RMG, mix for 5 minute.
2. Slowly add the granulation fluid to the RMG containing the Sifted water
extract of
stem of Tinospora cordifolia, stem plant powder over a period of about 3
minute,
with medium speed.
3. Stop the mixer and scrape off the mass from the sides and bottom.
4. Continue mixing by operating the impeller at high speed with Chopper ON for
about 3 minute.
5. Add additional quantity of purified water, if required.
6. Discharge the mass from the RMG.
WET MILLING
2 Mill the Wet Mass obtained in Multi mill fitted with 8 mm screen.
TRAY DRYING
1. Dry the Wet mass obtained in tray Drier at about 70 C to 80 C for about 60
minutes.
2. Check the Moisture once every 30 minutes.( LOD Limit: 2.0 to 5.0%w/w) and
record
the details
SIZING
1. Sift the dried granules using a Sifter fitted with sieve #16.
2. Collect the sifted granules in a clean double poly-lined HDPE container.
3. Mill the retains (oversize granules) obtained through a Multi Mill fitted
with 1.5 mm
screen with `Knives Forward' direction.
4. Pass the milled granules obtained through a Sifter fitted with #16 sieve.
5. Collect the sized granules obtained.
6. Weigh the granules.
a4^.
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7. Analyse samples as per below Table-24
Table-24.
Parameter Standard value
1 LOD 2.0-5.0%
2 BD 0.30 - 0.60 g/ml
3 Granules: Fine ratio 20:80 - 80:20
4 TVAC NMT 5000 cfu/gm
Fungal count NMT 10 cfu/gm
BLENDING
1. Transfer the sized granules to the blending area.
2. Transfer the sifted stem plant powder of Tinospora cordifolia lubricant to
the
blending area.
3. Load 5.0 kg of stem plant powder of Tinospora cordifolia and the sized
granules into
the double cone blender.
4. Blend the ingredients for about 6 minutes at 10-11 RPM.
5. Unload the blend in a clean Double poly-lined HDPE drums and affix duly
filled
status labels.
6. Weigh the blend and enter the details.
COMPRESSION
Maintain the temperature and relative humidity of the area with in the
specified limit (Limit
temperature NMT 25 C and relative humidity NMT 40%)
1. Check the Temperature and Relative humidity of the area and record.
2. Bring the drums containing the blend into the compression area.
3. Adjust the machine.
4. Carry the initial checks before starting the operation as specified in
below table-25
Table-25
Description
Punch size 17x8 mm caplet
Upper punch Plain
Lower punch Plain
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5. Check for Appearance, Average Weight, Individual Weight, Thickness,
Hardness,
and Friability & DT of 6 tablets.
6. Check appearance, thickness & hardness of tablets for every 30 min.
7. Check friability and DT every 1-hour
8. Check for the Average weight of 20 tablets every 30 mins graphically
9. Collect the tablets in a double poly-lined, tightly closed container. Weigh
each
container and enter the details.
STANDARD PARAMETERS FOR TABLETS:
Table-26
SI.No. Parameters Standard value
I Theoretical average weight 700 mg
2 Weight uniformity 700 mg 5% (665mg to 735 mg)
3 Weight of 20 tablets 14.00 g 3% (13.58 gm to
14.42 m)
4 Tablet thickness 3.5 to 6.5 mm
Tablet hardness 2 to 8 Kg/cm2
6 Friability NMT 1.0% W/W
7 Disintegration time NMT 30 min.
8 Total Bitters containing 24.6 mg per caplet
Tinosporaside
Tablets to be packed immediately after compression.
Packing
2. Pack 60 tablets in a HDPE 120 CC container and weigh them for
appropriateness of
number of tablets.
Example-30
LCMSMS STUDIES
Liquid Chromatography - Mass Spectrometer Analysis of Tinospora cordifolia
stem
supercritical (CO2) extract and water extract (Figure 7 & 8):
LCMSMS analysis were carried out by using an applied biosystem-Sciex API 2000
triple
quadrupole mass spectrometer equipped with an ion source turbo spray and ESI
interface.
The liquid chromatography was a LC-20 AD series binary system equipped with an
31
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autosampler. The column used was C18 phenomenex (250 X 4.6mm, 5 m), flow rate
0.5
ml/min of mobile phase methanol: water (0.1% acetic acid), (10:90), wave
length 254 nm and
run time 25 min. The analytes were ionized by ESI in positive-ion mode (PI
mode). Final
ionization conditions were heated ionization temperature 435 C. Curtain gas
Nitrogen 30 psi,
particulate -free and CO2- free air was used as ion source gas 1 at a flow
rate 50 psi and ion
source gas 2 at a flow rate 60 psi.
Sample preparation for Tinospora cordifolia stem CO2 extract:
Weigh about 50 mg of sample in a 50 ml volumetric flask, and dissolve by
sonication in
methanol (HPLC grade). Make the volume up to the mark with methanol filter
through 0.2um
syringe filter.
Sample preparation for Tinospora cordifolia stem water extract:
Weigh about 500 mg of finely powdered sample in a 50 ml volumetric flask, and
dissolve by
sonication in water (HPLC grade). Make the volume up to the mark with water
filter through
0.2 um syringe filter.
Example-31
Preparation of Ocimum sanctum water extract
Approximately 100 Kg of shade dried leaves of (Ocimum sanctum) was subjected
to
extraction with 400 Liters of purified water by percolation method at Room
Temperature.
The water extractions after 24 hours were filtered through muslin cloth and
concentrated to
thick paste. After achieving the desired total solid content, the soft extract
was spray dried to
a free flowing dry extract powder.
Example-32
Preparation of Ocimum sanctum SCFE (CO, extract
Approximately 25 Kg of leaves of Ocimum sanctum was pulverized to fine powder
and
loaded in to extractor. Super Critical Carbon dioxide liquid was pumped in to
the extractor at
a pressure of 350 bar and 40'C temperature for 2-4 hours. Extract was
separated into the
container at pressure of 40 bar and 20'C temperature. The CO2 super critical
liquid was
recycled from the extraction vessel.
3~
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Example-33
Formula of Granulation for Tulsi capsules (Table-27)
S. Name of the material Weight per Weight per
No Tablet in mg Batch in Kg
1 Ocimum sanctum leaves powder (Water extract) (# 40) 160.00 16.00
2 Ocimum sanctum leaves (Super critical fluid (CO2) extract) 60.00 6.00
3 Ocimum sanctum leaves (#80 mesh) powder 500.00 50.00
4 Purified water Granulation Quantity
fluid sufficient
Example-34
Formula of Lubrication of granules for Tulsi capsules (Table-28)
S. No. Name of the material Weight per Weight per
Tablet in mg Batch in Kg
1 Ocimum sactum Extract Granules (# 16 mesh) 720.00 72.00
E VEG CAPSULES CT/CT "00" WI/OUT
2 100000
LOGO, empty veg capsule
Fill weight 720.00 72.00
Example-35
Formula of Granulation for Tulsi tablets (Table-29)
S. Name of the Material Weight per Weight per
No Tablet in mg Batch In kg
Ocimum sanctum leaves powder (Water extract) (#
1 220.00 22.00
40) Dry extract
Ocimum sanctum leaves (Super critical fluid (CO2)
2 30.00 3.00
extract)
3 Ocimum sanctum leaves (#80 mesh) powder 400.00 40.00
4 Granulation fluid Quantity sufficient
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Example-36
Formula of Lubrication of granules for Tulsi tablets (Table-30)
1 Ocimum sanctum leaves granules (# 16 mesh) Granules 650.00 65.00
2 Ocimum sanctum leaves ( # 80 mesh) powder Lubricant 50.00 5.00
Total 700.00 70.00
Example-37
Manufacturing details of Tulsi Capsules
DISPENSING OF RAW MATERIAL
1. Dispense the raw materials as per Batch Formula.
DRY HEAT STERILIZATION
1. Transfer 50.0 kg of leaves powder of Ocimum sanctum in to trays and
sterilize the
material at 160 C for 60 mins
2. Unload the sterilized materials in to double lined polybags separately and
keep in
airtight containers.
3. Analyze the sample for LOD, BD and Microbiological Analysis Table-31
Table-31
S.No Parameter Standard values
1 LOD 2.5-3.5%
2 BD 0.30 - 0.40g/ml
3 Microbes NMT 10000 cfu/gm
4 Fungal count NMT 100 cfu/gm
SIFTING
1. Sift water extract of leaves powder of Ocimum sanctum through # 40 and
leaves
powder of Ocimum sanctum through #80, weigh as per the required quantity and
kept separately in duly-labeled double lined poly bag.
2. Record Quantity Sifted and the sieve integrity before and after sifting.
PREPARATION OF GRANULATION FLUID
1. Transfer about 20.0 kg of Purified water into a clean Stainless Steel
Vessel
2. Record the observations. Record deviations if any.
3y
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GRANULATION
1. Charge 50.00 Kg of leaves powder of Ocimum sanctum, add slowly 6.0 kg of
super critical extract (CO 2) of leaves of Ocimum sanctum into the RMG mix for
minutes.
2. Add 16.00 kg of water extract of leaves of Ocimum sanctum to the above
blend.
Mix it for 5 minutes.
3. Add granulation fluid to RMG and granulate. If required add additional
quantity
of purified water, mix over a period of about 3 minutes, with medium speed.
4. Stop the mixer and scrape off the mass from the sides and bottom.
5. Continue mixing by operating the impeller at high speed with Chopper ON for
I
minute.
6. Add additional quantity of purified water, if required.
7. Discharge the mass from the RMG.
8. Record the observations. Record deviations if any.
WET MILLING
1. Mill the Wet Mass in Multi mill fitted with 8 mm screen.
DRYING
1. Dry the Wet in FBD/Tray drier at about 55-60 C for about 60-90 minutes.
2. Check the Moisture once every 30 minutes. (LOD Limit: 2 - 4%) and record
the
details.
SIZING
1. Sift the dried granules using a Sifter fitted with sieve #18.
2. Collect the sifted granules in a clean double poly - lined HDPE container.
3. Mill the retains (oversize granules) through a Multi Mill fitted with 1.5
mm
screen with `Knives Forward' direction.
4. Pass the milled granules through a Sifter fitted with #16 sieve.
5. Analyze the sample.
6. In-process parameters are in Table-32
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Table-32
Parameter Standard values
1 LOD 1-2%
2 BD 0.60 - 0.70 g/ml
3 Granules to fine ratio 60:40 to 70:30
As per Finished Product spec
4 Actives
Microbes NMT 10000 cfu/gm
6 Fungal count NMT 100 cfu/gm
CAPSULE FILLING
Note: Maintain the Temperature and Relative Humidity of the area within the
specified limit
(Temperature 25 C 2 C and Relative humidity 40% 2 %)
1. Check the Temperature and Relative Humidity of the area and record.
2. Bring the drums containing the granules into the capsule filling area.
3. Adjust the machine.
4. Check for Appearance, Average Weight, Individual weight, average locking
length of 10 capsules& DT of 6 capsules.
5. Check DT every 1-hour.
6. Check Average weight of 20 capsules every 30 min graphically.
7. Collect the capsules in a double poly-lined, tightly closed, container.
8. Weigh each container and enter the details.
9. In process specification for capsules Table-33
Table-33
Parameter Standard limit
Description Clear Transparent size 00 capsules
filled with brown colored granules
Weight of empty capsules 115-120 mg
Fill weight 720 mg
Average weight 840 5%
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Weight 20 capsules 16.8 gm 3%
Average locking length of 10 capsules 23.3 0.4 mm
Disintegration NMT 15 min
8. Collect the filled capsules in double poly-lined HDPE drums and record
their
weights.
9. Affix duly filled status labels to the containers.
PACKING
1. Pack 60 capsules in a HDPE 120 CC container and weigh them for
appropriateness of number of capsules
Example-3 8
Manufacturing Details of Tulsi Tablets
Dry Heat Sterilization
1. Transfer the leaves plant powder of Ocimum sanctum into trays and sterilize
the
material at 160 C for 60 minutes.
2. Unload the autoclaved materials in to double lined polybags separately and
keep in
airtight containers.
3. Analyse sample for LOD, BD and Microbiological Analysis as per Table-34
Table-34
Parameter Standard values
1 LOD 2.0-3.0%
2 BD 0.35 - 0.55 g/ml
3 TVAC NMT 5000 cfu
4 Fungal count NMT 10 cfu
SIFTING
1. Sift water extract of leaves of Ocimum sanctum through # 40 mesh.
2. Sift leaves plant powder of Ocimum sanctum through # 80 mesh.
3. Collect the above-sifted materials in separate duly labeled double lined
polybags.
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PREPARATION OF GRANULATION FLUID
1. Transfer purified water into a clean Stainless Steel Vessel
GRANULATION
1. Charge of water extract of leaves of Ocimum sanctum and of leaves powder of
Ocimum sanctum into the RMG, mix for 5 minutes.
2. Slowly add super critical extract (CO2 extract) of leaves of Ocimum
sanctum, and
mix for about 3 minutes.
3. Add granulation solution to the above blend and mix for about 3 minutes,
with
medium speed.
4. Stop the mixer and scrape off the mass from the sides and bottom.
5. Continue mixing by operating the impeller at high speed with Chopper ON, if
required.
6. Add additional quantity of Purified water, if required.
7. Discharge the mass from the RMG.
WET MILLING
1. Mill the Wet Mass obtained in Multi mill fitted with 8mm screen.
TRAY DRYING
1. Dry the Wet mass in tray Drier at about 70 C to 80 C for about 60 minutes.
SIZING
1. Sift the dried granules using a Sifter fitted with sieve #16.
2. Collect the sifted granules in a clean double poly-lined HDPE container.
3. Mill the retains (oversize granules) through a Multi Mill fitted with 1.5
mm
screen with `Knives Forward' direction.
4. Pass the milled granules obtained through a Sifter fitted with #16 sieve.
5. Collect the sized granules obtained and add them to the sifted granules.
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BLENDING
Granules were analysed as per table-35
Table-35
Parameter Standard values
1 LOD 3.0-4.0%w/w
2 BD 0.35 - 0.55 g/ml
3 TVAC NMT 5000 cfu
4 Fungal count NMT 100 cfu
Maintain the Temperature and Relative Humidity of the area within the
specified limit
(Limit Temperature NMT 25'C and relative humidity NMT 40%)
1. Transfer the sized granules into the blending area.
2. Transfer the sifted leaves powder of Ocimum sanctum lubricant into the
blending
area.
3. Load 5.0 kg of leaves powder of Ocimum sanctum and the sized granules into
the
Double Cone blender.
4. Blend the ingredients for 5 minutes at 10-15 RPM.
5. Unload the blend in a clean Double poly-lined HDPE drums and affix duly
filled
status labels.
6. Weigh the blend and enter the details.
COMPRESSION
Maintain the Temperature and Relative Humidity of the area with in the
specified limit (Limit
0
Temperature NMT 25 C and relative humidity NMT 40%)
5. Check the Temperature and Relative Humidity of the area and record.
6. Bring the drums containing the blend into the compression area.
7. Adjust the machine as per the parameters.
8. Carry out the initial checks before starting the operation as specified in
below
Table-36
Table-36
Description
Punch Size 17x 8 mm caplet
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Upper Punch Plain
Lower Punch Plain
10. Check for Appearance, Average weight, Individual weight, Thickness,
Hardness, Friability & DT of 6 tablets.
11. Check appearance, thickness & hardness of tablets every 30 min.
12. Check Friability and DT every 1-hour.
13. Check Average weight of 20 tablets every 30 mins graphically.
14. Collect the tablets in a double poly-lined, tightly closed, container.
Weigh each
container and enter the details.
STANDARD PARAMETERS OF TABLETS
Standard parameters are mentioned in Table-37
Table-37
S. No. Parameters Standard value
1 Theoretical average weight 700 mg
2 Weight uniformity 700 mg 5% (705mg to 695mg)
3 Tablet thickness 4.0 to 5.0 mm
4 Tablet hardness 4 to 6 Kg/cm
Friability NMT 1.0% W/W
6 Disintegration time NMT 30 min.
7 Total Oleonolic acids 3.5 mg per caplet
12. Collect the Compressed tablets in Double poly-lined HDPE drums and record
their weights.
13. Affix duly filled status labels to the containers.
PACKING
14. Pack 60 tablets in a HDPE 120 CC container and weigh them for
appropriateness of number of tablets.
Example-39
Liquid Chromatography -Mass Spectrometer Analysis of Ocimum sanctum leaves,
supercritical (CO?) extract and water extract (Figure 9 and 10)
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LCMSMS analysis were carried out by using an applied biosystem-Sciex API 2000
triple
quadrupole mass spectrometer equipped with an ion source turbo spray and ESI
interface.
The liquid chromatography was a LC-20 AD series binary system equipped with an
autosampler. The column used was C18 phenomenex (250 X 4.6mm, 5 m), flow rate
0. 5
ml/min of mobile phase methanol: water (0.1% acetic acid), (10:90), wave
length 254 nm and
run time 25 min. The analytes were ionized by ESI in positive-ion mode (PI
mode). Final
ionization conditions were heated ionization temperature 435 C. Curtain gas
Nitrogen 30 psi,
particulate -free and C02- free air was used as ion source gas I at a flow
rate 50 psi and ion
source gas 2 at a flow rate 60 psi.
Sample preparation for Ocimum sanctum leaves CO2 extract:
Weigh about 50 mg of sample in a 50 ml volumetric flask, and dissolve by
sonication in
methanol (HPLC grade). Make the volume up to the mark with methanol filter
through 0.2
um syringe filter.
Sample preparation for Ocimum sanctum leaves water extract:
Weigh about 500 mg of finely powdered sample in a 50 ml volumetric flask, and
dissolve by
sonication in water (HPLC grade). Make the volume up to the mark with water
filter through
0.2 um syringe filter.
While this invention has been described in detail with reference to certain
preferred
embodiments, it should be appreciated that the present invention is not
limited to those
precise embodiments. Rather, in view of the present disclosure, which
describes the current
best mode for practicing the invention, many modifications and variations
would present
themselves to those skilled in the art without departing from the scope and
spirit of this
invention.
Abbreviated terms for following.
1) LOD Loss on drying
2) BD Bulk density
3) RMG Rapid Mixer Granulator
4) FBD Fluid Bed Drier
5) TVAC Total Viable Aerobic Count
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6) NMT Not More Than
7) DT Disintegration Time
4a
