Journal of Pharmacognosy and Phytochemistry 2022; 11(3): 250-259

E-ISSN: 2278-4136
P-ISSN: 2349-8234
www.phytojournal.com Formulation and evaluation of polyherbal
JPP 2022; 11(3): 250-259
Received: 16-03-2022 capsules containing combination of Terminalia
Accepted: 24-04-2022
arjuna, Chrysanthemum indicum and Moringa
Shubhangi Bhide
Department of Pharmacognosy,
oleifera
Career Point University, Kota,
Rajasthan, India
Shubhangi Bhide and Dr. Vikas Jain
Dr. Vikas Jain
Department of Pharmacognosy, Abstract
Career Point University, Kota,
Natural medicine, especially from herbs, is the source for the research of various novel medicinal
Rajasthan, India
compounds. Drugs from herbal origin must be ensured as safe before used as medicine.
Objective: The present work focused on the formulation, development and evaluation of polyherbal
capsule containing ethanolic extracts of Bark of Terminalia arjuna, flowers of Chrysanthemum indicum,
leaves of Moringa oleifera.
Methods: The ethanolic extract of all three plants were taken for the formulation and combination of all
three extract was selected for the formulation of the capsule. The combination of three ethanolic extracts
(CTEE) which include Bark of Terminalia arjuna, flowers of Chrysanthemum indicum and leaves of
Moringa oleifera was selected as a sample material. The material was weighed, sampled, authenticated
and analyzed for their compliance to quality standards. Evaluation of the capsules was done based on
different parameters.
Results: Preliminary phytochemical screening of CTEE revealed the presence of major phytochemical
groups such as alkaloids, carbohydrates, tannins, steroids and sterols, triterpenoids, saponins and
flavonoids. As per the standards, the flow property of the blend to be filled in the capsules should be in
good range and was confirmed by the above parameters. Trial batch- 3 showed excellent flow characters
and that batch was taken for capsule filling. Physical parameters Moisture content-3.6%±0.22,
Uniformity of weight-268 mg ± 4.5mg, Disintegration time-3.32 (min) ± 0.34, pH(1% aqueous solution)-
7.33 ± 0.21.
Conclusion: To enhance the acceptability of the herbal medicine by consumers, many of the products
have been formulated into conventional dosage forms such as tablets, capsules, suspensions, and
powders. CTEE were used in this study to formulate a unit solid dosage form (capsule) to increase the
compliances, acceptability and adaptation of the consumers. As it is also very important to estimate the
pharmaceutical quality of the Herbal products irrespective of their medicinal content and therapeutic
states; so in the present study, the pre-formulation and formulation studies of the formulated capsules.
Absorption of drug in the blood is controlled by the availability of drug from solid dosage into the GI
fluid. Hence the rate of absorption and availability may be improved by improving the disintegration and
the rate of dissolution of drug. Capsule has been successfully formulated.

Keywords: Terminalia arjuna, Chrysanthemum indicum, Moringa oleifera, capsules etc.

Introduction
Hypercholestermia is a life-threatening disorder that develops through elevated lipids content
in the blood circulation. Lipids play a vital role in the body’s muscle growth, but an abnormal
level of fats in the blood highly increases the risk factor for developing coronary heart
diseases. Nowadays, cardiovascular diseases are a serious life-threatening epidemic disorder in
India [1]. Cardiovascular diseases are responsible for one-third of the total deaths worldwide,
and it is believed that cardiovascular diseases will prove to be a leading cause of morbidity and
mortality in forthcoming years [2].
Hyperlipidemia is caused by the elevation of total cholesterol, triglycerides, very low density
lipoprotein, and low density lipoprotein in plasma. Hyperlipidemia is also caused by a
decreased level of high density lipoprotein in blood. Hyperlipidemia with an elevated level of
lipoproteins is measured by the initiation and progression of plaque formation in arteries which
may causes thrombosis and myocardial infraction [3]. Control and reduction the lipid level is
Corresponding Author:
necessary for freedom from coronary artery diseases. However, the drug therapies using
Shubhangi Bhide niacin, clofibrate, gemfibrozil, atorvastatin, cholestyramine, cholestipol and probucol
Department of Pharmacognosy, administered for the treatment of hyperlipidemia may produce an unexpected toxic effect [4].
Career Point University, Kota, Probucol especially was withdrawn due to its undesired side-effect of lowering HDL levels
Rajasthan, India
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and QT interval prolongation in patients with a previous United States contain at least one active ingredient derived
history of heart diseases. Consequently, herbal rutin and its from plant material. Some are made from plant extracts;
compound used for the treatment of hyperlipidemia have been others are synthesized to mimic a natural plant compound [9].
approved since it has no undesirable side-effects, its use is
economic and easily available [5]. Experimental work
Hyperlipidemia is a disease of lipid metabolism produced by Chemicals and reagents
elevation of plasma concentration of the diverse lipid and Ethanol 99.9% was procured from LOBA Chemicals,
lipoprotein fractions, which are the source of cardiac disease. Mumbai. Ethylene di-aminetetraacetic acid (EDTA) was
It is define as increase serum TC, TG, VLDL, LDL and HDL procured from Thermo Fisher Sci- entific India Pvt. Ltd.,
which are responsible for different complications like: heart (Mumbai, India). All the sol- vents used were of high purity
attack, coronary artery syndrome, stroke, atherosclerosis, and HPLC grade. All other chemicals and reagents used in the
myocardial infarction and pancreatitis. Hyperlipidemia can be whole study were of analytical grade.
either primary or secondary type, the primary syndrome may
be treated by hypolipidemic drugs, but secondary induced by Pharmacognostic evaluation of plant materials
diabetes, hypothyroidism or renal lipid nephrosis which Collection and authentication of plant material
treated by treating the original disease moderately than The Bark of Terminalia arjuna, flowers of Chrysanthemum
hyperlipidemia. Genetic disorders and lifestyle diet rich in indicum and leaves of Moringa oleifera was collected locally
calories, fat, and cholesterol play a vital role to cause in the Thane district. The plant materials were then
dyslipidemia around the world [6]. The main factor which are authenticated from Ideal College of Pharmacy and Research,
responsible for hyperlipidemia includes changes in life style Kalyan.
habits in which risk factor is mainly poor diet i.e. fat intake
greater than 40 percent of total calories, saturated fat ingestion
more than 10 percent of total calories; and cholesterol
ingestion larger than 300 milligrams per day [7]. For
hyperlipidemia large number of synthetic drugs available, not
a bit is helpful for all lipoprotein disorders, and each drugs are
linked with a number of adverse effects.

Classification of hyperlipidemia Preparation of extracts

Hyperlipidemia may be classified as either familial (also The collected plant material (Bark of Terminalia arjuna,
called primary) caused by definite genetic abnormalities, or flowers of Chrysanthemum indicum and leaves of Moringa
acquired (also called secondary) that leads to change in oleifera) (500 g) each was gently washed by using distilled
plasma lipid and lipoprotein metabolism. water to remove the impurities. The collected materials were
Familial (primary): -Familial hyperlipidemia is classified as: shade dried in the laboratory under room temperature (24 ± 2
Type I: Raised cholesterol with high triglyceride °C) for 3–4 weeks. After complete drying, the dried plant
Type II: High cholesterol with normal level of triglyceride material was and pulverized by using a mechanical grinder
Type III: High cholesterol and triglycerides followed by sieving to obtain a coarse powder. The powdered
Type IV: Raised triglycerides, and raised uric acid plant material was then extracted with distilled water and
Type V: Raised triglycerides ethanol (99.9%) using reflux technique separately. Extracts
were concentrated by vacuum distillation and then dried in
Herbal medicine open air to produce the respective extracts. The crude aqueous
Herbal Medicine sometimes referred to as Herbalism or and ethanol extracts obtained was stored at 4 °C before
Botanical Medicine, is the use of herbs for their therapeutic or analysis. The percentage yield of the extract was calculated by
medicinal value. An herb is a plant or plant part valued for its the following formula.
medicinal, aromatic or savory qualities. Herb plants produce Percentage yield = (Weight of dry crude extract
and contain a variety of chemical substances that act upon the obtained/weight of plant material before extraction) X 100
body [8]. The weight in gram was used to calculate the percentage
Herbal medicine is the oldest form of healthcare known to yield. The calculated percentage yield was as follows;
mankind. Herbs had been used by all cultures throughout
history. It was an integral part of the development of modern Percentage
No. Extract
yield
civilization. Much of the medicinal use of plants seems to
1 Aqueous extract of Bark of Terminalia arjuna 7.24%
have been developed through observations of wild animals, 2 Aqueous extract flowers of Chrysanthemum indicum 6.34%
and by trial and error. As time went on, each tribe added the 3 Aqueous extract leaves of Moringa oleifera 4.48%
medicinal power of herbs in their area to its knowledgebase. 4 Ethanol extract of Bark of Terminalia arjuna 8.54%
They methodically collected information on herbs and 5 Ethanol extract flowers of Chrysanthemum indicum 5.35%
developed well-defined herbal pharmacopoeias. Indeed, well 6 Ethanol extract leaves of Moringa oleifera 7.78%
into the 20th century much of the pharmacopoeia of scientific
medicine was derived from the herbal lore of native peoples. Phytochemical screening
Many drugs commonly used today are of herbal origin. The Phytochemical screening was done by the standard
Indeed, about 25 % of the prescription drugs dispensed in the procedure as depicted in Table:

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Table 1: Preliminary Phytochemical Screening

Chemical constituents Chemical test
Proteins Biuret test
Molish test
Carbohydrates
Fehling’s test
Dragendorff’s test
Alkaloids
Mayer’s test
Salkowaski test
Steroids
Liebermann-burchard test
Vanillin-sulphuric
Triterpene
acid test
Ferric chloride test
Tannins
Dilute nitric acid test
Glycosides Keller-killani test
Shinoda test
Flavonoids
Lead acetate test
Saponins Foam formation test
Amino acids Ninhydrin test

Preformulation studies Preservatives

To formulate any dosage forms, it is essential that The preservatives are added to herbal formulation to prevent
fundamental physical and chemical properties of the drug contamination, deterioration and spoilage by bacteria, fungal
powder are to be determined [66, 67]. and other microorganisms. The most effective preservatives
are the sodium methyl paraben, sodium propyl paraben,
Definition sodium benzoate and bronopol.
Preformulation involves the application of biopharmaceutical Selection of excipients in the formulation are given below
principles to the physicochemical parameters of drug  Talc - Glident/Lubricant
substance are characterized with the goal of designing  Microcrystalline cellulose - Diluent/Disintegerant
Optimum drug delivery system. Before beginning the  Starch - Binder/Disintegerant
Preformulation programs the preformulation scientist must  Colloidal sillicon dioxide - Glident
consider the following factors  Magnesium stearate - Lubricant
 The amount of drug available.  Bronopol - Preservative
 The physicochemical properties of the drug already  sodium methyl Paraben - Preservative
known.
 Therapeutic category and anticipated dose of compound. Preparation of formulation
 The nature of information, a formulation should have or  The dry CTEE (Combination of Three Ethanolic
would like to have Extracts) were dried in tray drier at 60 oc for 20 minutes.
All excipients used in this formulation except
Selection of excipients preservatives were dried separately in tray drier at 100oc
For the formulation of capsules in addition to the active for 30 minutes. All active ingredients were weighed
ingredients, excipients like diluents (filler), binder, according to the formula, mixed and lubricated with
disintegrating agent, lubricant and preservatives are required. magnesium stearate followed by diluents and
The choice of excipients was made keeping in mind the preservatives were mixed well. The mixture was blended
current Food and Drugs Administration (FDA) regulations. thoroughly for 30 minutes. Then the powder was
[68]
. transferred to the polythene bags and labelled for further
studies [115].
Diluents
Diluents/Fillers are added where the quantity of active Development of formulation-trial batches
ingredient is less (or) difficult to filling. Common Three trial batches were formulated by varying the
tablet/capsule filler include Lactose, Dicalcium phosphate, composition of the excipients proportions for excellent flow
microcrystalline cellulose, etc. properties.

Lubricants Table 2: Development of formulation

They reduce friction during the filling process. In addition,
they aid in preventing adherence of capsule material. No. Materials Trial-1 (g) Trial-2 (g) Trial-3 (g)
1 CTEE 20 20 20
Magnesium Stearate, Stearic acid, Hydrogenised vegetable
2 Talc 1.7 1.8 2.5
oils and talc are commonly used lubricants.
3 Mcc 0.8 0.9 1
4 Starch 2 2.5 2.6
Glidants 5 Magnesium stearate 0.8 1 1.5
It is used to improve flow of the powder materials by 6 Colloidal sillicon dioxide 0.25 0.28 0.3
reducing the friction between the particles. The most effective 7 Bronopol 0.15 0.15 0.15
glidants are the Colloidal silicon dioxide, Talc and Starch. 8 Sodium methyl paraben 0.15 0.15 0.15

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Evaluation of blended powder Capsules are small dosage form in which one or more
The blended powder of all trial batches were analysed for its medicinal and inert ingredients are enclosed in a small shell
flow characteristics like bulk density, tap density, usually made of gelatin.
compressibility index, Hausner’s ratio and angle of repose.
Capsule Size and Selection of Filling Method
Bulk density and tap density and Carr’s index  The formulated granules were filled in “1” size capsules
A weighed quantity w of (15g) powdered material was taken to an average net content t weight of 270 mg.
in a 50 ml measuring cylinder. And recorded the initial  The capsules were then de dusted, transferred into
volume (vo) tapped the contents and recorded the powdered polybags, labelled and the Samples were evaluated as per
volumes after 50 taps (v50). the testing requirements.
The formula for fluff density and Tapped density is as follow  A hand operated gelatin capsule filling machine was used
Fluff density = w/vo g/cc in this study for encapsulation of capsules.
Tapped density = w/vο 50 g/cc  From the final trial, samples were taken for accelerated
The formula for Carr’s index is as follow stability studies as per the testing requirements.

Carr’s index = {Tapped density- Fluff density/ Tapped

density} X 100
Value for Carr’s index below 15 indicate excellent flowing
material and value over 20-30 suggested poor flowing
material.

Angle of repose
A funnel was fixed at a particular height (1.5, 2.5, 3.5 cm) on
a burette stand. A white paper was placed below the funnel on
the table. The powdered drug passed slowly through the
funnel until it forms a pile. The radius of the pile was noted
down. Angle of repose of the powder material was calculated
by using the formula:

Tan θ = h/r,θ = tan (h/r) where, h = height of the pile, r =

radius. Fig 2: Herbal capsule

Values for angle of repose < 30ο usually indicate a free Standardisation of herbal capsules [70-75]
flowing material and angle > 400 suggest a poor flowing The developed herbal capsules were standardized for its
material. description, uniformity of weight, disintegration time,
moisture content, physicochemical parameters, phytochemical
Hauser’s ratio studies, fluorescence analysis. Standardization were carried
The basic procedure is to measure the unsettled apparent out as per Indian pharmacopoeia procedures.
volume, V0 and the final tap volume Vf , of the powder Following Quality control parameters were evaluated
tapping the material until no further volume changes occur.
The Hausner’s ratio was calculated as follows: 1. Description
Hausner’s ratio = V0/Vf The general appearance of a capsule, its visual identity and
overall “elegance” is essential for consumer acceptance. The
Hausner’s ratio between 1.00 to 1.11 shows excellent flow color, shape, odor and surface texture are all noted for the
and value more than1.60 shows very poor flow. capsules prepared.

Formulation of capsules [77-82] 2. Uniformity of weight

From the 3 trial batches one optimized batch is selected for 20 individual units were selected at random and their content
formulation based on above results. Trial batch 3 was found was weighed and their Average weight was calculated. Not
to be the perfect batch and it was selected for the more than two of the individual weights deviate from the
consideration of further evaluation. average weight by more than the percentage shown in the
table
Table 3: Final batch composition -250 mg/capsule
Table 4: Acceptance Criteria I.P Limit
No Ingredients Quantity in mg
1. CTEE 200 Dosage form Average limit Deviation
2. Micro crystalline cellulose 12.5 < 300 mg 10%
3. Starch 7.4 -10 capsules
>300mg 7.5%
4. Sodium methyl paraben 0.25
5. Talc 25 3. Disintegration test
6. Colloidal sillicon dioxide 3.7 Disintegration test was performed using the digital
7. Magnesium stearate 0.85 microprocessor based disintegration test apparatus. One
8. X-Bronopol 0.25
capsule was introduced into each tube and added a disc to

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each tube. The assembly was suspended in the water in a 1000 effect at non acceleration and to evaluate the effect of short –
ml beaker. The volume of water was such that the wire mesh term excursions outside the label store conditions such a
at its highest point is at least 25 mm below the surface of might occurs during shipping. Result from accelerated testing
water, and at its lower point was at least 25 mm above the studies are not always predictive of physical changes.
bottom of the beaker. The apparatus was operated and
maintained the temperature at 37±2 °C. (Indian Conditions of Stability studies
Pharmacopoeia, 2010).  Accelerated condition of 40 °C ± 2 °C/75% RH ± 5% RH
 Long term condition of 25 °C ± 2 °C/60% RH ± 5 % RH
4. Determination of moisture content  Long term / intermediate condition of 30 °C ± 2 °C/75 %
The loss on drying test is important when the herbal substance RH ± 5% RH
is known to be Hygroscopic. An excess of water in medicinal
plant materials will encourage Microbial growth, the presence The ICH Harmonized Tripartite Guideline provides a general
of fungi, insects deterioration. In modern Pharmaceutical indication on the requirements for stability testing of new
technology, the water content provides information drug substances and products. The main thrust of the stability
concerning the Shelf life and quality of the drugs. guideline centers on criteria for setting up stability protocols.
Moisture content (%) = {Final weight of the sample/Initial
weight of the sample} ×100 6. Climatic zones
The four zones in the world that are distinguished by their
5. pH characteristic prevalent annual climatic.
1 g of capsule powder was taken and dissolved in 100 ml
demineralized water. The pH value of the solution was Table 5: Climatic zones and derived storage conditions
determined by means of a digital pH meter. The pH meter was Zone Condition Temperature Humidity
calibrated using buffers of 4, 9 and 7 pH. The electrodes were Zone 1 Temperature 21ºc 45% RH
immersed in the test solution and pH was measured. Zone 2 Sub-tropical with possible Humidity 25ºc 60% RH
Zone 3 Hot/ dry 30ºc 35% RH
Accelerated stability studies of the capsules Zone 4 Hot / humid 40ºc 70% RH
Stability is defined as the extent to which a product retains,
within specified limits and throughout its period of storage Accelerated stability condition : Accelerated stability study
and use (i.e., its shelf-life) the same properties and were carried out of storage condition at 40 °C ± 2 °C of
characteristics that it possessed at the time of its manufacture. humidity 70% RH for 3 month(time period covered).

6. Accelerated testing Result and Discussion

Studies designed to increase the rate of chemical degradation Preliminary phytochemical screening
or physical change of a drug substance or drug product by Preliminary phytochemical screening of CTEE revealed the
using exaggerated storage conditions as per of the formula presence of major phytochemical groups such as alkaloids,
stability studies. Date from the studies, in addition to long tern carbohydrates, tannins, steroids and sterols, triterpenoids,
stability studies, can be used to assess longer term chemical saponins and flavonoids as shown in Table.

Table 6: Preliminary phytochemical screening of three selected plant

Chemical Bark of Terminalia arjuna Flowers of Chrysanthemum indicum Leaves of Moringa oleifera
Chemical test
constituent Ethanolic extract Ethanolic extract Ethanolic extract
Proteins Biuret test + + +
Molish test + + +
Carbohydrate
Fehling’s test + + +
Dragendorff’s test - - +
Alkaloid
Mayer’s test - - +
Salkowaski test + - -
Steroids
Liebermann-burchard test + - +
Vanillin-sulphuric
Triterpene + - +
acid test
Ferric chloride test - + +
Tannin
Dilute nitric acid test - + +
Glycoside Keller-killani test + + -
Shinoda test - + +
Flavonoid
Lead acetate test - + +
Saponins Foam formation test - - +
Amino acids Ninhydrin test + - -

Phytochemical study reveals the presence of some organic Result of polyherbal formulation capsule
compounds which provide definite physiological action on the The combination of three ethanolic extracts (CTEE) which
human body and these bioactive substances include tannins, include Bark of Terminalia arjuna, flowers of
alkaloids, carbohydrates, terpenoids, steroids, flavonoids, and Chrysanthemum indicum and leaves of Moringa oleifera was
saponins. selected as a sample material. The material was weighed,

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sampled, authenticated and analyzed for their compliance to Results (n-=3) are reported as mean ± standard deviation.
quality standards as established by WHO guidelines,  1% aqueous solution of herbal formulation showed acidic
pharmacopoeial and other standard reference books. pH.
 The average weight of the capsules was calculated as per
Preformulation Studies I.P and the obtained value was with in the limit (±10%).
Three trial batches of the herbal formulation were prepared  Sample were taken randomly (3times) to specify
and tested for Preformulation parameters like bulk density, quantity, the moisture content was calculated as per trail
tap density, Carr’s index, Hausner’s ratio and Angle of and error by KFR titration method. The result were given
repose. The results observed is shown in table. in the above table.
 Disintegration time of the herbal capsule was performed
Table 7: Final batch composition -250 mg/capsule
as per I.P and the obtained value showed that the capsule
No Ingredients Quantity in mg will be disintegrated within the prescribed time for the
1. CTEE 200 absorption.
2. Micro crystalline cellulose 12.5  The uniformity of weight of the capsules was calculated
3. Starch 7.4 -10
as per the I.P and obtained value was within limit (±7.5).
4. Sodium methyl paraben 0.25
5. Talc 25  The formulated herbal capsule weight were the lower
6. Colloidal sillicon dioxide 3.7 limit is noted as 248 mg and the upper limit is noted as
7. Magnesium stearate 0.85 287mg.

Table 8: Evaluation of in process parameters The rate and extent of absorption of a drug into the
bloodstream is an important quality characteristic of a dosage
Parameters Trial-1 Trial-2 Trial-3
form. In vivo bioavailability and in vitro dissolution studies
Bulk density (g/cm) 0.42±0.01 0.38±0.05 0.35±0.04
Tap density (g/cm) 0.45±0.03 0.47±0.01 0.50±0.04 are important in the development and ultimately in the quality
Compressibility index(%w/w) 26.83±0.66 23.26±2.54 13.06±1.12 control of a dosage form. Formulation studies involve
Hausner ratio 1.35±0.15 1.22±0.02 1.13±0.01 developing a preparation of the drug which is both stable and
Angle of repose 40.42±2.57 39.36±2.67 34.66±0.18 acceptable to the patient. For orally taken drugs, this usually
involves incorporating the drug into a tablet or a capsule. The
All values are expressed as standard mean deviation ±, where n=3 medicine derived from plants can be used more conveniently
and safely in various diseased conditions, if used in proper
Table 9: Evaluation of in process parameters portions and combination. To enhance the acceptability of the
Parameters Trial-1 Trial-2 Trial-3 herbal medicine by consumers, many of the products have
Flow property Normal Fair Perfect been formulated into conventional dosage forms such as
Filling Uniform Uniform Uniform tablets, capsules, suspensions, and powders. CTEE were used
Weight Not uniform Not Uniform Uniform in this study to formulate a unit solid dosage form (capsule) to
Moisture content Satisfied Satisfied Perfect increase the compliances, acceptability and adaptation of the
Disintegration time Within the limit Within the limit Perfect consumers.

As per the standards, the flow property of the blend to be Conclusion

filled in the capsules should be in good range and was As it is also very important to estimate the pharmaceutical
confirmed by the above parameters. Trial batch- 3 showed quality of the Herbal products irrespective of their medicinal
excellent flow characters and that batch was taken for capsule content and therapeutic states; so in the present study, the pre-
filling. formulation and formulation studies of the formulated
The trial 3 flow properties were Excellent and all parameter capsules. Absorption of drug in the blood is controlled by the
were within the Specified limits. So, third trial was chosen for availability of drug from solid dosage into the GI fluid. Hence
further studies. the rate of absorption and availability may be improved by
improving the disintegration and the rate of dissolution of
Standardisation of finished formulation drug. Capsule has been successfully formulated. The
The final batch was tested for organoleptic characters, formulated Dosage forms met the pharmacopeia criteria for
physical and physico chemical parameters. The results quality Assessment and can be used as suitable alternatives in
observed are shown in table the Management and treatment of hyperlipidemia.
Table 10: Organoleptic characters
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