Formulation and Standardization of Asava from Carica papaya

 

Rohile V. Y.1*, Patil V. M.2, Patil S. S.2, Desai A. V.1, Inamdar N. R.3

1Ashokrao Mane Institute of Pharmacy, Ambap, Kolhapur, Maharashtra, India, 416112.

2Ashokrao Mane College of Pharmacy, Peth-Vadgaon, Kolhapur, Maharashtra, India, 416112.

3Annasaheb Dange College of B Pharmacy, Ashta, Sangli, Maharashtra, India, 416301.

*Corresponding Author E-mail: rohilevinayak@gmail.com

 

ABSTRACT:

Asava and Arishta are alcoholic medicaments prepared by allowing the herbal juices or their decoctions to undergo fermentation with the addition of sugar. Standardization of ayurvedic formulation is important so as to assess the standard of medication. Within the present study standardization of asava from fruit tree Carica papaya, known to be effective in Dengue fever, Cancer cell growth inhibition, Antimalarial has been performed. Asava formulation was prepared by the normal method of Ayurveda. The formulation has been standardized by modern scientific control procedures for the finished products. Standardization of asava was achieved by organoleptic study, physicochemical parameters like PH, denseness, total solid content, amount, alcohol content, index of refraction, total reducing sugars, and stability study. The results have revealed that the physicochemical parameters were within the bounds and also the values may be wont to establish and formulate procedures for standardization and quality controlling of those ayurvedic formulations.

 

KEYWORDS: Asava, Standardization, Carica papaya, Dengue fever.

 

 


INTRODUCTION:

Ayurveda is an ancient Indian medical system dating back to the Vedic period about 3000-1500 BC. The word Ayurveda is composed of two parts ayur and veda, Ayur means life and veda means knowledge. India is having a rich heritage of traditional medicine constituting with its different components like Ayurveda, Siddha and Unani. Botanical constituents are the major part of these traditional medicines. The development of these traditional systems of medicine with the perspectives of safety, efficacy, and quality will help not only to preserve the traditional heritage but also to rationalize the use of natural products in healthcare. Ayurveda is considered as oldest healing science. In Sanskrit, Ayurveda means “The Science of Life”.1 Asava and Arishta are alcoholic medicaments prepared by allowing the herbal juices or their decoctions to undergo fermentation with the addition of sugars.2

 

Formulation of Asava and Arishta has done by soaking the drugs, either in coarse powder form or in the form of decoction, in a solution of sugar or jaggery, for a specified period of time, during which it undergoes a process of fermentation.3,4 Asava it is liquid preparations containing self genareted alcohol, thus contain water soluble as well as alcohol soluble substances of the drugs. Due to their medicinal value, sweet taste, and easy availability people are prone to consume higher doses of these drugs for longer periods. As per Samhita Asava is Madya which is prepared with apakwa Aushada. The compound which is prepared by ‘asutaprakriya’ is called as Asava.5,6 Asava and arishta i.e. sadhanakalpna are considered to be the unique and best dosage form discovered by Ayurveda. Asavas are prepared by the fermentation of herbal juices and arishtas are prepared by the fermentation of the decoction of plants.9 The preparation of asava involves the mixing of the powdered drugs, jaggery or honey or both in plant juices or water whereas in arishtas a decoction of some plant ingredients is prepared, filtered, cooled and mixed with powdered drugs, honey or jaggery or both.7 Vagbhatahas also defined Asava as Madya prepared using fresh tubers, roots, fruits etc. and Madya with medicinal properties is called as Asava.8,9 The aim of present study was formulation and standardization of the Asava an Ayurvedic formulation to establish typical quality control parameters of such formulations.

 

MATERIALS AND METHODS:

Materials:

All powdered ingredients were collected from Unique chemicals, Kolhapur, Maharashtra, India. All Chemicals used were of analytical grade.

 

Methods:

Approximately 250ml of water are taken in to the cleaned fermentation flask (glass; 350ml), with hot water and Sugar/Jaggary/Wheat floor, was dissolved in to it. This mixture of water and sugar source was boiled for half an hour, cooled to room temperature to have a final mixture. All ingredients as given in table 1 were weighed accurately and passed through sieve no 44 and ingredients from 1 to 14 were added to the final mixture. Dhatakipusha (Woodfordiafructicosa) was then added to the mixture. The vessels was closed with a cleaned lid followed by wrapping around the lid with clay smeared cloth. The vessel was kept at dark place for 40 days.11

 

Table 1: Formulation Table

S. No

Ingredients

Formulation code

A

B

C

1.

Papaya powder

750 mg

750 mg

750 mg

2.

Neem powder

225 mg

225 mg

225 mg

3.

Karela powder

225 mg

225 mg

225 mg

4.

Gudmar powder

300 mg

300 mg

300 mg

5.

Triphala powder

1950 mg

1950 mg

1950 mg

6.

Belpatra powder

225 mg

225 mg

225 mg

7.

Haridra powder

225 mg

225 mg

225 mg

8.

Chirayta powder

225 mg

225 mg

225 mg

9.

Lodhra powder

150 mg

150 mg

150 mg

10.

Shatawari powder

225 mg

225 mg

225 mg

11.

Dalchini powder

150 mg

150 mg

150 mg

12.

Tulsi powder

150 mg

150 mg

150 mg

13.

Babul powder

150 mg

150 mg

150 mg

14.

Water

150ml

150ml

150ml

Prakeshpa Dravya

 

 

1.

Dhatakipushpa

2000 mg

2000 mg

2000 mg

2.

Sunth powder

150 mg

150 mg

150 mg

3.

Sugar

40 g

-

-

4.

Jaggery

-

40g

-

5.

Wheat flour

-

-

40g

 

After the stipulated period, the vessel was withdrawn. Preparation showed dark brownish fluid, pleasant smell and alcoholic taste. The fluid was filtered through muslin cloth, prepared asava was kept in a container.

 

Standardization of Asava:

Preliminary Evaluation of Asava Formulations:

Organoleptic characteristics odour, taste, colour and clarity of prepared asava formulation was determined.

 

Ph:

Digital pH meter was used to check the pH of formulations. Before the experiment the machine was calibrated by using standard buffer solution of Ph 4.0, 7.0 and 9.2.

 

Determination of Density and Specific gravity:

The density and specific gravity were determined by using density bottle method.10

 

Determination of viscosity:

The viscosity was determined by Ostwald viscometer. The viscosity was determined by formula.10

 

n2= (ρ2t2 / ρ1t1) x n1

Where   

ρ2 =    Density of water

ρ1 =  Density of sample

t1 =  Time required for water

t2  =  Time required for sample

n1 =  Viscosity of water.

n2 =  Viscosity of sample.

 

Determination refractive index:

Abbe’s Refractometer was cleaned and prisms were wipe out with alcohol and then with acetone. Instrument was kept near to light source, and placed water in prisms with help of capillary tube. After that eye piece was adjusted such a that, boundary at the center of the cross wire, and reading was taken as refractive index of water. Same procedure fallowed for the test sample that is asava formulation.

 

Total solid content determination:

In evaporating dish 20ml of the Asava formulation was taken and evaporated to dryness on a water bath and then dried at 1050C for 3 hours. The obtained residue was kept in a desiccator for 30 min, and weighted. The total solid content was determined by following formula.5

 

Total solid content = Weight of dish after drying - weight of empty dish

 

Ethanol content by distillation and specific gravity:

Preparation being examined was transferred about 25ml, to the distillation flask. It was diluted with 150ml of water and to it added a little pumice powder. Till not less than 90ml of the distillate was collected it was distilled. The distillate was diluted with distilled water. The relative density and alcohol content was determined as per Ayurvedic pharmacopeia.5

 

Determination of total reducing sugars:

5ml of fermented broth was pipette with clean and sterilized pipette and diluted with 40ml of distilled water to which, 3ml of concentrated HCL was added. The mixture was shaked well and heated to boil. After cooling to room temperature neutralized with anhydrous sodium carbonate. It was further diluted with distilled water up to 100ml and titrated against 10ml of Fehling’s solution (5ml Fehling’s ‘A’ and 5ml Fehling’s ‘B’). Methylene blue was used as indicator and end point was change in color from deep blue to the red of the cuprous oxide. The calculation was done by following formula.5

 

% TRS = 5.128 / B.R × F.F × D.F

TRS – Total reducing sugar

B.R- Burette reading

 

Stability study P:

Stability study was carried out at 400C and 75% Rh for 30, 60 and 90 days for observing changes in physicochemical nature in all prepared formulations of asava. The stability chamber used was REMI SC-6 PLUS.10

 

RESULTS AND DISCUSSION:

Preliminary Evaluation and organoleptic properties:

The preliminary evaluation of Asava is as given in table 2

 

 


Table 2: Preliminary evaluation of Asava

Days of Fermentation

Colour

Odour

Touch

Taste

0

Light brown

Slight alcoholic

Watery

Sweet, sliight bitter

5

Light brown

Slight alcoholic

Watery

Astringent bitter

10

Brown

Alcoholic

Watery

Astringent bitter

15

Brown

Alcoholic

Watery

Astringent bitter

20

Dark brown

Alcoholic

Watery

Astringent

25

Dark brown

Alcoholic

Watery

Astringent

30

Dark brown

Fragrant

Watery

Astringent

35

Dark brown

Fragrant

Watery

Astringent

 


Asava was observed dark brownish in colour, fragment aromatic odour, astringent taste and watery touch after minutes in texture between fingers. At initial stage of fermentation color was observed light brown and taste was sweet but slightly bitter. There was no frothing or release of gases. Asava formulations were with characteristics bitter taste. No sediment was found.

 

The organoleptic evaluation of asava is as given in table 3


 

Table 3: Organoleptic evaluation of Asava

S. No

Parameters

Batch A

Batch B

Batch C

1.

PH

3.44 ± 0.03

3.62 ± 0.04

4.82 ± 0.04

2.

Specific gravity

1.077 ± 0.002

1.092 ± 0.004

1.306 ± 0.004

3.

Density

1.023 ± 0.002 gm/ml

1.031 ± 0.004 gm/ml

2.042 ± 0.41 gm/ml

4.

Viscosity

1.043 ± 0.004 cp

1.042 ± 0.002 cp

1.0182 ± 0.005 cp

5.

Refractive Index

1.344±0.002

1.349±0.014

1.375±0.002

6.

Total solid

20.494 ±1.322 % w/v

29.126 ±0.992 % w/v

31.698 ±1.157 % w/v

7.

Alcohol content

5.331           ± 0.577 % v/v

5.669± 0.577 % v/v

1.331±0.577 % v/v

8.

Acid value

1.627±0.115

1.353±0.147

1.33±0.148

9.

Total reducing sugar

8.11 ± 0.52 %

7.91 ± 0.679 %

10.331 ± 0.218 %

10.

Rf Value

0.74

0.70

0.71

 


 

PH, Specific gravity, Density, and Viscosity:

The PH of the formulations was analyzed to evaluate acidity of the formulations. The formulations were found acidic in nature. The pH of the formulations A, B and C after the fermentations were found to be 3.44±0.03; 3.62±0.04 and 4.82±0.04 respectively. Generally, formulations having less alcohol content have higher pH. Hence from the result it has been revealed that A and B were well fermented as compared to C. It has indicated that sugar and jiggery should preffered to be better for fermentation than wheat flour.

 

Specific gravity and density is the most important evaluation test for checking completion of the fermentation. The specific gravity and density of the formulation were found to be A: 1.077±0.002, 1.023 ±0.002gm/ml; B: 1.092±0.004, 1.031±0.004g/ml; C: 1.306±0.004, 2.042±0.41g/ml. respectively, values indicated that fermentation is complete.

The viscosity of the Asava formulation was found to be 1.043±0.004; 1.042±0.002 and 1.018±0.005cp, respectively of batches A, B and C.

 

Refractive index, Total solid content, Alcohol content:

The Refractive Index of prepared Asava formulation A, B and C was found to be 1.344±0.002, 1.349±0.014 and 1.375±0.002 respectively. Total solid content have moderate impact over viscosity. The total solid content of the formulation A, B and C was found to be 20.494± 1.322 % w/v; 29.126±0.992% w/v and 31.698±1.157% w/v. The alcohol content (average) was found to be 5.331±0.577% v/v; 5.669±0.577 % v/v and 1.331±0.577 % v/v of batches A, B and C respectively.

 

Acid value, Total reducing and Rf Value:

The acid value was found to be 1.627±0.115, 1.353±0.147 and 1.33±0.148 of batch A, B and C respectively, Total reducing sugar content was determined by fehlings method. The total reducing sugar from the formulation A, B and C was found to be 8.11±0.52 %; 7.91±0.679 %; 10.331±0.218%. Thus, it has indicated that for better reproducibility and to have optimized quality above parameters should be used as comparative tool.

 

Stability Study:

The stability parameters study of Asava are as given in table 5. Stability studies were performed at accelerated Condition 450C Temperature and relative 75% humidity (RH) for 90 days. No change was noticed in colour, odor and taste of Asava in all batches for 90 days. Slight changes were noticed in pH of both batches at accelerated temperature during storage.

 

Table 4: Stability study of asava

Parameter

Time

Observations

A

B

C

Colour

0

Dark brown

Dark brown

Dark brown

30

Dark brown

Dark brown

Dark brown

60

Dark brown

Dark brown

Dark brown

90

Dark brown

Dark brown

Dark brown

Inference

NC

NC

NC

Odour

0

Alcoholic Fragrant

Alcoholic Fragrant

Alcoholic Fragrant

30

Alcoholic Fragrant

Alcoholic Fragrant

Alcoholic Fragrant

60

Alcoholic Fragrant

Alcoholic Fragrant

Alcoholic Fragrant

90

Alcoholic Fragrant

Alcoholic Fragrant

Alcoholic Fragrant

Inference

NC

NC

NC

Taste

0

Astringent

Astringent

Astringent

30

Astringent

Astringent

Astringent

60

Astringent

Astringent

Astringent

90

Astringent

Astringent

Astringent

Inference

NC

NC

NC

pH

0

3.44

3.62

4.82

30

3.41

3.63

4.80

60

3.40

3.63

4.80

90

3.39

3.62

4.79

Inference

SC

SC

SC

Sugar Content

0

8.11%

7.91%

10.331%

30

8.10%

7.88%

10.330%

60

8.08%

7.87%

10.329%

90

8.08%

7.88%

10.329%

Inference

SC

SC

SC

NC: Not changed      SC: Slightly changed

 

Changes during Fermentation:

Changes observed during fermentation as shown in figure 1, 2 and 3 for pH, specific gravity and total solid content respectively.

 

Figure 1: Change in pH during fermentation

 

Figure 2: Change in specific gravity during fermentation.

 

Figure 3: Change in Total solid content during fermentastion.

 

Evaluation were caried out after every 5 days. In case of pH initialy was slight neutral which turns to acidic after fermentation in all batches. Specific gravity was 1.02, 1.386 and 1.411 which was reduced 1.077, 1.107 and 1.311; also total solid content was reduced after fermentation.

 

CONCLUSION:

The current study of formulation and it’s standardization of asava by physicochemical methods is a new approach and an attempt to add on the number of analytical methods by which an ayurvedic formulation such asava can be conveniently subjected for it’s quality control, unlike adopting few process such as determination of alcohol and sugar content alone. All evaluation parameters should be used as quality control tool for assuming quality and for batch-to-batch uniformity. From the comparative analysis of laboratory and marketed product, it can be confirmed that there is uniformity in preparation of formulations. The results obtained from the study could be utilized as a reference for setting limits for the reference standards for the quality control and quality assurance of such formulation. Hence, we concluded that this study will be a referential tool for attempting various methods of analysis to standardize Ayurvedic formulation (specifically for Asava) which may be taken as quality control parameter for future study.

 

ACKNOWLEDGEMENT:

The authors are thankful Principal, Ashokrao Mane college of pharmacy, Peth-Vadgaon for providing the necessary facilities for carrying out the experimental work.

 

CONFLICT OF INTEREST:

No.

 

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Received on 24.04.2020            Modified on 30.05.2020

Accepted on 26.07.2020         © RJPT All right reserved

Research J. Pharm. and Tech. 2021; 14(4):2211-2215.

DOI: 10.52711/0974-360X.2021.00392