INTRODUCTION

The history of Indian Pharmaceutics is as old as history of Ayurveda.

Historically Ayurveda might have been brought on the earth along with

mankind. Ayurveda gives highest importance to maintenance of health

and to promote positive health rather than to cure disease. Due to

various reasons healthy process of life is disturbed and diseases are

produced and cause suffering to mankind.

At this stage, drugs play an important role along with other healthy

principles in removing the suffering of mankind. But drugs in their

original form could not be used internally either for maintenance of

health or for prevention and cure of diseases. These have to be

processed and convert in to some suitable pharmaceutical forms, which

on internal use may absorb and assimilated and produce their desired

effect without causing any toxic manifestations. The knowledge

regarding drugs, their processing techniques, their pharmacological

and therapeutic properties and method of their uses comes under the

preview of pharmaceutical science.

Bhaisajya Kalpana is the branch of Ayurveda, where in standard raw

drugs is transferred in to a potent medicine. Acharya Caraka and

Vagbhata have given importance to Dravya after Bhisak.

Ancient sage and scholars of Ayurveda have always tried to make the

preparations more palatable and potent. Keeping the prepared medicine

for longer duration they yielded a fermentating procedure called

SandhanaKalpan6. There are many fermentated preparations found

mentioned in Ayurvedic Pharmaceutics since ancient times.

Asava and Arista are popularly used by Ayurvedic practitioners since

many centuries. These preparations occupy prime position in Indian

Pharmacopoeia on account of their superiority to other preparation.

They are fast acting and can be stored for long time without losing

its potency.

Pindasava is a classical Asava-arista product of Ayurveda mentioned

to treat Grahani, which is a common clinical problem in every place.

The Pindasava has proven its action in many cases, its action is

evaluated clinically but the Pharmaceutical and standardization of

this drug not yet evaluated.

Standardization assures that those products are reliable in terms of

quality, efficacy, performance and safety. It helps to avoid

adulteration and improper substitution. The aim of this study is to

fix the physico-chemical standards of Pindasava and the drugs used in

the formulation. Each drug is characterized on the basis of their

intrinsic chemical constituents these determination in the formulation

and the variation in batch to batch preparations.

The standardization of herbal drugs and their formulation will be

effective when the classical and modern mentioned methods used in

combination, each methods playing a complementary role. There is a

growing need to apply these methods to standardize the herbs and their

formulations to find global acceptability.

Pindasava is a special type of Asava preparation which is described in

Caraka Samhita in the context of Grahanicikista. This product is

special in its physical characteristic has the name indicated

Pindasava is in Pinda or semisolid form, unlike other asava arista

which are in liquid form. This preparation even though these claim to

be very efficacy in managing on important disease Grahani, is not

being prepared regularly or used in clinical practice. This may be

because of the non-availability of standard operative procedure of

Pindasava hence difficulty in preparation. Keeping these in mind it

was decided to practically prepare Pindasava as per textual reference

its batches along with pharmaceutical modification in the intention to

develop standard operative procedure and also in-house quality control

standards.

CONCEPTUAL STUDY

This included the review of literature and drug review.

While reviewing the literature an attempt was made to look for the

textual reference of Pindasava, it was found that Pindasava is

referred in Carakacikitsa 15th chapter while explaining Grahanicikitsa,

here elaborated procedure of preparation is not given. Here the

ingredient are mentioned and quantity of pippali and jala is specified

but not of the entire ingredient. Later in Asava arista Vijnanam,

Vaidya Paksadhara Jha explained the ingredients, quantity and

procedure of preparation Pindasava. Vaidya Kru. Su. Bhatta in Kannada

book Asava arista Vijnanam also has elaborated the procedure of

Pindasava.

Considering all these references, present study was planned. According

to all the available references, Pippali and Vibhitaki mentioned are

being chief ingredient of Pindasva in addition Guda and Jala are used.

As per the reference Madhyama Vibhitaki mentioned Carakacikitsa stana,

Vibhitakabeeja Majja which forms the central portion of fruit was

taken. Pindasava is the formulation considered under Asava arista

Kalpana which are the outcome of Sandhana process hence in literary

review concept of sandhana Kalpana with special reference to Asava

arista was compiled elaborated and analysed.

HISTORICAL REVIEW

The knowledge of Sandhana Kalpana starts from Vedic period. The

process of fermentation (Sandhana),

SAMHITA PERIOD

Charaka Samhita :

Theory along with practical applications (pharmaceutically and

clinically) and detailed information regarding Sandhana Kalpana51 are

mentioned in this text. In Sutrasthana 15th chapter Siddhu, Sura,

Sauviraka etc. are mentioned as fermented preparations.

9 Asava Yoni (sources), 84 Asava drugs along with the properties of

Asava are mentioned in 25th chapter of Sutrasthana.

Specific type of pots : Regarding place : Regarding duration :

Use of metal :

Use of metal in Arista formulation is also found.

e.g. Triphaladhyarista

Sushruta Samhita :

Madya Varga of Sutrasthana in 45th chapter deals with the 27 types of

fermented preparations.

Astanga Hridaya :

In this text, Madya Varga is described in the 5th chapter of

Sutrasthana

MEDIEVAL PERIOD

Chakradatta :

Total 5 Asava Arista preparations are mentioned with method, in one of

the formulations the author has used Dhatki for Lepana e.g.

Dantyarista.

Gada Nigraha : mentioned 60 Asava Arista

Sharangadhara Samhita :

Acharya Sharangadhara (13th century) has established the fundamental

principles of every Kalpana (formulations).

Difference and definition of Asava Arista is given in the text which

is strictly followed in the formulation of Asava Arista. Total

thirteen Asava Arista are mentioned, among them four are Asava and

nine are Arista.

Other

Other nighantus like Bhaisajya Ratnavali, Yogaratnakara , Rasa tantra

sara Shodhala Nighantus, Raja Nighantu, Nghantu Ratnakara,

Dhanwantari Nighantu etc have also mentioned about sandhana Kalpana.

Sandhana is classified on various parameter on the basis of sourcematerial used i.e. Asava yoni- 84 Asavadravya mentioned across 9

Asava yoni. On the basis of natures of final product obtained

Madyavarga and Sukthavarga are the classification mentioned. The

product where Alcohol is generated is categorized in Madyavarga and

products forming organic acid like Acetic acid are classified under

Sukthavarga. These categories have been further classified by

AcharyaYadavji.

Shukta Varga :

The products which attains Amlata (sourness), after fermentation

process is known as Shukta. Amlata (sourness) is desirable in Shukta

Sandhana. Ex; tusambu, kanjika, Dhanyamla.

 

Shukta (Amlata) is achieved by two preparations.

Direct acidic fermentation

Through alcoholic fermentation

Fermentation carried out by Acid bacilli.

By acidification process, ethyl alcohol converts into acetic acid

which is an oxidative process. Sourness in Asava Arista (Madya) is a

consequence of contamination with bacteria (mainly acetobactor) which

is capable to convert alcohol to acetic acid.

Madya Sandhana (Alcoholic fermentation)

Asava- usually with sita virya drugs, preffered to pittika vyadi,

preference to pediatrics

Arista – usna veerya drugs, preferred to Vatha-kaphaja vyadi,

preferred in adult and old age.

In case of Madyavarga sugar or carbohydrate converted into Alcohol by

the process of anaerobic fermentation along with the production of

carbon dioxide. This process is accelerated by the action of enzymes

like (present in sugar yeast) Saccharomyces cervisiae.

Fermentation in Modern view :

Drug review .

The mode of preparation, its uses and its therapeutic dose explained

in the classical text, Charakasamhithachikitsasthana, 15th Chapter

Grahanidhosacikitsa adhyaya.

One prastha each of pippali(Piper longum, Linn.) jaggery and seed-pulp of

bibhithaki (Terminaliabelerica, Roxb) should be made to a paste and added

with one prastha of water. This should then be kept in a jar and

covered with the grains of barley till it is well-fermented. This is

called Pindasava. One pala of this medicated alcoholic drink should be

mixed with one anjali of water and taken. This recipe cures all the

diseases. Even a healthy person is advised to take this proportion for

one month in order to prevent the occurrence of these diseases.

Pippali

Vibhitaki beeja majja

Guda

Dhataki puspha.

Pharmaceutical Study

This is the key part of present research project, Here Pindasava was

prepared as per textual guide line and also with practical

modification to develop Standard Practical Operative Procedure for

Pindasava. The task was highly challenging for following reasons.

Formulation is unique with extremely high solid content making

the wort semisolid,

No reference of previous work carried out is available.

Market samples prepared by pharmaceutical company are also not

available.

Considering these difficulties and uncertainty of getting the product,

properly prepared a Pilot study was planned initially.

Pilot Study:

Pindasava was prepared in 3 batches.

First batch was exactly as per classical reference.

In second batch the quantity of water was doubled.

Third batch triple the quantity of water was taken.

The process was followed methodically after Sandhibandana the changes

where clearly observed.

In a classical method as the quantity of water too less when compared

to the other, the granules were produced. No since of Fermentation

like Effervescence, Alcoholic odour could be seen. A thick layer of

white mold with black powder covered the sample on 15th day. Sample

emitted bad odour and sour taste.

The product as considered spoiled and discarded.

In a sample where double the quantity of water used the product was

like thick paste, here also fungus developed on 15th day with the

development of bad odour and taste.

The product had to be discarded.

Third sample with the triple quantity of water was thick liquid it

shown signs of fermentation after 30 days. Where effervescence could

be seen at the end of two month mild alcoholic odour could be

appreciated but it shows white fungul growth the taste was not

palatable and the product had to be discarded.

By the pilot study it could be learnt that product get contaminated

quickly and fungus was developed. Quantity of water in classics

reference is too less for yeast growth and fermentation. A delay in

Initiation of fermentation looks to be the prime reason for the

spoiling of the product.

On the basis of this experience standardized preparation was planned

with suitable precaution as much as possible aseptic precautions taken

to prevent contamination.

Samples were heated for about 15 minutes to remove or to destroy any

microbial contamination. Dhatakipuspha was used in 3 of the sample to

enhance fermentation process. To regulate the temperature

sandhanapaatra were kept inside grains and husk.

For this study 6 sample of Pindavasa were prepared. 3 sample same as

pilot study but with adequate precaution and three other samples were

the same but with the addition of Dhataki. Dhataki was found to be

source of contamination in a number of studies to avoid this

dhatakijala was prepared as per the experience of Vaidya Dhamantkar

and used.

Another modification made in this study was in the quantity of

ingredients used. As the liquid quantity is less in the classical

reference and difficulty carried out fermentation process let to take

the individual ingredient by volume instead of by weight.

After mixing the ingredient the pH of water that was 7 changed to 5.8

after the process of boiling.

In the first sample i.e. B1 as per the classic, a sample was brown

thick like avaleha, initially on 7th day slight effervescence could be

seen on the surface, slight darkening of the sample. At the end of 30th

day mild alcoholic odour was appreciable with positive lime water

test. It takes 144 days for the completion of fermentation.

In second sample B2 where same classical formulation was added with

Dhataki puspa jala shows almost similar observation but the process

looks to be slightly faster and fermentation got completed in 130

days.

In sample B3 where double quantity of water used than classical used

fermentation initiated 7 day which was more vigorously at end of 30th

day. Here also fermentation got completed at 130th day.

Sample B4 was same as B3 except of addition of Dhatakipuspa jala ,

here comparatively fermentation process was farter and got completed

in 124 days.

Similarly Pindasava was prepared by using triple quantity of water

than classical reference with and without addition of Dhatakipuspa

jala , it was observed that Pindasava B5 where only triple quantity of

water used fermentation was completed in 110 days.

Least time is taken in the fermentation in the sample where triple

quantity of water and Dhatakipuspa jala was used; here fermentation

was completed in 96 days.

Above observation made it clear that minimum quantity of water is

required for proper fermentation. Dhataki places definite role in the

fermentation process. Preheating helps to destroy the microorganisms.

Previous study has all ready confirmed the role of Dhatakipuspa in

fermentation. Further Das; showed that the flower of Dhataki contains

substantially high concentration of Tannins to the extents of 22% and

such polyphenolic compound susceptible to enzymatic conversion to

simple Phenols and alcohols during anaerobic fermentation of arista

preparation. Perhaps just justifies the extensive uses of Woodifordia

Fruticosa (Dhataki) in arista preparation in the main purpose of which

is produce alcohol128.

There is also opinion that an endogenous invertise

(Fructofuranosidase) found in Dhataki flower, helps in sucrose

hydrogen to alcohols. The alcohol production helps in promoting the

extraction of biological active component including Gallic acid which

is otherwise traces if at all, as well as Ayurvedic process was ‘self

generated alcohol’.

Insinuates a conjecture here the researchers have tried to proved that

Dhatakipuspa is effective component not only for initiate the

fermentation but also enhancing clinical efficacy129.

A lesser duration required for fermentation, smooth completion of

fermentation observed in present study in sample with Dhatakijala.

Further support the earlier report.

Analytical Study

Analysis of pharmaceutical product by following classical and advanced

modern methods of analysis is very important for the standardization

and quality control. Hence this forms an important part of study in

present research.

Here an attempt was made to analysis all the six sample of Pindasava

with the help of Organoleptic and physico- chemical method of

analysis. Followed by advanced HPTLC technique to get an idea about

their Physico chemical character phytoconstituents and quality

standard for Pindasava could be developed.

Organoleptic Analaysis: All the samples Keenly observed for

organolepthic characters both before and after completion of

fermentation.

Pindasava sample B1 (as per classic) was semisolid with brownish black

colour and strongly odour of pippali. Sample B3 with double quantity

of water was thick paste with moderate odour of pippali & dark brown

colour. Sample B5 with triple quantity of water was dark brown in

colour but lighter than other 2 sample. It had the odour of pippli but

comparatively lesser extends than other sample. Constitency was thick

liquid due to the higher amount of water.

Similar observations were seen in samples B2, B4, B6 containing

Dhataki Jala.

After the completion of fermentation Pindasava sample of B2 with that

is classical had same semisolid constituency & dark brownish dark

colour, odour was significantly changed, intensity of aroma of Pippali

was reduced, faint odour of vibithaki could be appreciable & odour was

present. In addition to Madhura & Katurasa, Kasaya Rasa is

appreciated.

In the sample pindasava B3 along with odour of pippali, vibithaki seed

mild Alcoholic odour could be appreciated slight acidic taste appeared

in product. In sample B5 Alcholico dour were more prominent this

product had Kashaya and Amla Rasa in equal intensity in addition to

Madhura & Katu.

Sample Pindasava B2 in classical ratio with Dhataki Puspa Jala was

pleasant odour along with aroma of Pippali and Vibithaki Bheeja. Here

also Kashaya Rasa appeared.

The product was cold to touch where asall the sample without dhataki

puspa jala that is B1 B3 B5 where warm. Cold touch is probably with

generation of higher amount of Alcohol. Pindas of sample B4 had Amla

Rasa &Kashaya Rasa in addition to Madhura and Katu. It was colder to

touch. Pindasava sample B6 had more intense of Alcoholic order and

coldest touch among the entire sample.

When compiled to the alcoholic percentage generated it’s clear that

coldest of the product and alcoholic order are proportional to the

amount of Alcohol generated.

PH – All the samples were slightly acidic i.e. between 5.5 to 6

before fermentation. A progressive reduction of pH is observed at the

end of the 30th day, 60th day and at the end of completion respective

days.

This reduction in pH is because of development of alcohol and sour

taste. Higher the alcohol in the sample lower was the pH.

Specific gravity :

The sample were semisolid to thick liquid in consistency hence weight

of 25 ml of sample was taken and mass was recorded on this basis of

specific gravity was calculated. Specific gravity is the ratio of

density of the substance to the density (Mass of the same unit

volume) of the reference substance.

Apparent specific gravity is the ratio of the weight of the volume of

the substance to the weight of equal volume of reference substance. In a presence study specific gravity was found to decrease

progressively as a volume of water added was increased. There was no

difference in pattern among the sample with or without dhataki pushpa

jala. It is also found that Alcohol percentage is increased specific

gravity reduced gradually.

Refractive Index:

The ratio of a substance with reference to air is the ratio sine of

angle of incidence to the sine of to the angle of refraction of beam

passing from air to the substance. Previous studies have shown that

determination of refractive index. That is one of the best suiting

standardization processes for liquid formulation with reference to

air130.

Hence in the presence study refractive index was calculated for all

the samples. A slight reduction was observed in refractive Index. As

the dilution increased in the sample hence ratio increases of samples

5 & 6 where lowest at 1.4285 & 1.4446 respectively.

Ref. Index is directly related to brix value & hence a similar pattern

also seen in brix value of sample also.

Brix value – brix value indicates sugar concentration in a sample

hence the brix can be used for soluble sugar evaluation hence it

represent amount of soluble sugar in the product pindasava. When the

result where observed maximum brix value was observed for the sample

prepared as per the classics which is followed by the samples with

classical ratio as per the additional of dhatakipushpajala. In other

sample as dilution increases birx value formed reduced naturally.

Total solids : In the presence total solid was calculated in 50 ML

sample in grams usually total solid in a liquid formulation given the

amount of solute that is soluble material in a unit volume. But here

as filtration not done the entire mass is considered for estimation.

Hence the naturally as the dilution of the sample increases total

solid content as found to be reduced.

Alcohol content: Alcohol was calculated by the process of distillation

specific gravity of distillate were compared with a table to get the

alcoholic content in percentage.

Estimation of Alcohol content is very important parameter for

Asava Arista preparation it indicates several factors like adequacy

of Fermentation yeast activity, sugar conversation, preservation etc.

Alcoholic generated acts as preservative and also help for

Facilitation of active principle from the ingredients. It was found

that a minimum quantity of Alcohol was generated in all the

formulation, lowest (4.5) being in classical sample concentration of

alcohol are found to be increased along with dilution. Hence with the

double quantity of water (6.5) and with triple quantity (7.5) of

alcohol were observed.

Amount of Alcohol was marginally higher when Dhatakipushpa jala was

used. This was uniformly observed in all the sample (5,7, 7.5%

against 4.5, 6, 7.5 %)

Extractive value :

Extractive value indicates amount of the material soluble in a

specific solvent. This is specific for specific type of drug or

formulation. Hence it is used as parameter for standardization of

herbal product.

Water soluble extract also followed similar pattern as produced

reduction in value was observed as dilution increased.

Quantitative test for Flavonoids, Steroids and Saponins:

Quantitative test for Phytoconstituents is a important parameter for

standardization. It depend upon the chief constituent present in the

product. Qualitative test were conducted as standard protocol all the

samples showed the present of flavoniods and steroids.

Saponins could not be detected.

Previous studies have indicated that the chief constituent of the

product Pindaasava that is Vibhitaki131 and Pippali132 both contents

flavonoids and steroids. This may be the reason for the presence of

flavonoids and steroids in all the samples of Pindaasava.

Reducing and non-reducing sugar:

Sugar is the source of alcohol in asava arista preparations during the

process of anaerobic fermentation. Sugar gets converted in to alcohol

due to action of yeast along with the production of carbon dioxide.

Hence estimation of sugar is important for quality detecting

parameter. Initially sugar contain of guda the sweetening agent of

Pindasava were estimated. It had total sugar of 51.9% of which

reducing sugar is 33.6% and non-reducing sugar is 18.19%.

Reducing sugar mainly indicate soluble carbohydrates. Non-reducing

sugar mainly includes sucrose 133. During the process of fermentation

total sugar get reduced as alcohol is generated which is clearly seen

in a present study.

In the present study fermentation reduction of non-reducing sugar

observed as alcohol content is increased. The fast reducing sugar is

espected because it includes sucrose, which is not fermentable by

yeast, but after enzymatic transformation on reducing sugar, (glucose

and fructose) by yeast invertase can be converted in to ethanol. This

is because alcoholic fermentation is bio chemical reaction here yeast

transfer the sugar (glucose and fructose) from the wort in to ethanol,

carbon dioxide and energy storage as ATP134.

Similar pattern of conversion of sugar observed earlier study135 also.

Hence in a present study Pindasava sample B5 and B6 which add maximum

alcoholic percentage as 7.5% shown a list amount of non-reducing sugar

that is 4.85 and 3.91% respectively.

Test for Methanol

Methanol is simple alcohol but highly toxic if induced. It is usually

produced by process of oxidation, it may be formed from ethanol when

it is called denatured spirit. Its consumption can damage optic nerve

even at a small quantity and higher dose proved to be fetal. Hence

test for methanol is important to assure quality and safe of asava

arista preparation. When tested none of the sample of Pindasava shown

the presence of methanol.

Chromatography:

Chromatography is technique of separation of individual component from

a mixer of compounds. It forms a very important tool for

standardization of herbal product.

Thin layer chromatography is simple at very useful technique for the

quality assessment of herbal products. It is useful for qualitative

and semi quantitative estimation of phyto constituent in an herbal

products. It also forms a base for further advance technique like

HPTLC, HPLC analysis.

For the present study methanol extract of sample where spoted along

with methanol extract of chief ingredient Pippali churna and Vibhitaki

bija Majja churna. Chromatogram was developed by using toluene; ethyl

acetate are 9:1 as per the TLC method for Pippali explained in API.

After developing the Chromatogram when visualized under visible light

shows two greenish spot with R.F value 0.046 and 0.36.

Neither Vibhitaki nor any of the the sample showed any spot.

When visualized under long wave UV (366nm) Pippali shows three pinkish

spotes with RF value 0.04,0.16,0.26.

Two flurosent pink spotes with RF value 0.34,0.63.

Vibhitaki did not show any spotes.

All the sample showed spot with RF value 0.63 corresponding to the

component present in pippali.

Under short UV light Pippali shown three pinkish spot with RF value

0.266,0.34 and 0.45.

Vibhitaki did not show any spot.

Pindasava prepared as per the classical ref. showed pinkish spot with

RF value 0.21,0.28 and 0.34 other sample shows spot with RF value

0.23,0.33,0.39.

Majority of the spot have been mentioned in standard TLC protocol of

Pippali136using the same solvent system in API vol.4.

The present study indicate that TLC is very useful parameter for

quality standardization of Pindasava.

HPTLC study:

HPTLC is a very useful for standardization of herbal product it

consumes only small amount of solvent system and needs only short

period for development and can evaluate multiple sample very quickly.

It generates finger prints which help for batch to batch quality

control. Here quantitative estimation of compound is possible when

compound is known and pure marker is available.

HPTLC analysis of Pindasava for Gallic acid content Vibhitaki137, as an

important marker Gallic acid which also process immune’s therapeutic

value.

As Pindasava contain Vibhitaki as chief ingredient and attempt was

made to quantitatively estimate gallic acid content in Pindasava

sample. Even after following standard protocol none of the sample

showed the presence of gallic acid this is most probably the reason

that gallic acid is reachly present in Vibhitaaki fruit pulp not in

seed. In present study Vibhitakibijamajja was used for preparation of

Pindasava.

HPTLC analyses of Pindasava sample for piperine content.

Another important ingredient of Pindasava is Pippali which were well

known marker piperine. Piperine was observed in TLC study also. Mobile

phase containing tolene: Ethyl acetate: methanol(8.5:1.5:0.25) was

used .RF value for piperine for this solvent system was 0.4 to ± 0.01

When evaluated. All the representative of the asava sample showed the

presence of piperine contents which valued between 0.2 to 0.38% w/w.

Interestingly Pindasava preparations showed maximum percentage piper

0.38%.

Quantitative estimation of alkaloid and tannin:

Alkaloids are present in both Pippali and Vibhitaki138 used in chief

ingredient of Pindasava.Tannin are present in Vibhitaki

By considering this report and estimation of alkaloid (chiefly

piperine) and also tannin quantitatively was planned. All the samples

tested showed the presence of alkaloid and tannin. Total alkaloid

present was almost similar in all the sample varying between 0.050 and

0.558.

Tannin contain are also similar among the sample varying between

0.0057 to 0.0069%, the maximum being in the classical sample.

Estimation of total fat in Pindasava sample

In the present study Vibhitakibijamajja was chiefly ingredient of the

Pindasava sample. Seed kernel Terminalia bellarica139containe about 43%

of oil. This oil contains 32.8% palmitic acid and 31.3% oleic acid ,

28.8% linooleic acid.

Hence an attempt was made to estimation of total fat contains of

Pindasava sample. All the test sample showed the presence of fat

varying between 1.72 to 2.28%.

Hence this can be quality parameter for standardization of Pindasava

sample.

Microbial analysis of Pindasava sample

According to WHO guide lines and CCRAS guide line for microbial

contamination in herbal product is mandatory. All the samples taken

passed the standard limit of all the type of microorganism

contamination tested. This ensures that Pindasava prepared by all the

experiments methods are safe and devoid of harmful microbial

contamination.

Conclusion:The sample with no fermentation initiator, indicates lower alcohol

content proved the importance of Dhataki puspa in fermentation..

Observation made it clear that minimum quantity of water is required

for proper fermentation.

Preheating helps to destroy the microorganisms

By the pilot study it could be learnt that product get contaminated

quickly and fungus was developed. Quantity of water in classics

reference is too less for yeast growth and fermentation.

As the liquid quantity is less in the classical reference and

difficulty carried out fermentation process let to take the individual

ingredient by volume instead of by weight.  

Organoleptic Character of the sample of Pindasava are slightly

different in each batch. Which may be due to role of water quantity,

fermentation initiators and alcohol contents.

Pindasava prepared with triple quantity of water found to be faster

fermentation with more alcoholic content.

Pindasava Prepared with Dhataki puspa jala shows early fermentation

with pleasant odour this observation confirm that the role of

Dhatakipuspa in fermentation.

TLC and HPTLC suggests the presence of alkaloid of Pippali.

HPTLC for Gallic acid content shows, none of the sample showed the

presence of gallic acid this is most probably the reason that gallic

acid is reachly present in Vibhitaaki fruit pulp not in seed. In

present study Vibhitaki beeja majja was used for preparation of

Pindasava.

Al the samples taken passed the standard limit of all the type of

microorganism contamination tested. This ensure that Pindasava

prepared by all the experiments methods are safe and devode of harmful

microbial contamination.