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 Table of Contents  
ORIGINAL ARTICLE
Year : 2022  |  Volume : 16  |  Issue : 2  |  Page : 126-133

Physico-chemical standardization and nutritional assessment of Devdarvadyarishta


1 Department of Rasa Shastra and Bhaishjya Kalpana, Saraswati Ayurved Hospital and Medical College, Gharuan, Mohali, India
2 Department of Rasa Shastra and Bhaishjya Kalpana, Faculty of IMS, BHU, Varanasi, Uttar Pradesh, India
3 Department of Rasa Shastra and Bhaishjya Kalpana, Rishikul Campus, UAU, Haridwar, Uttarakhand, India

Date of Submission14-Jan-2021
Date of Decision24-Nov-2021
Date of Acceptance11-Feb-2022
Date of Web Publication4-Jul-2022

Correspondence Address:
Chinky Goyal
Saraswati Ayurved Hospital and Medical College, Gharuan, Mohali - 140413, Punjab
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/joa.joa_14_21

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  Abstract 


Introduction: Devdarvadyarishta is a honey based anti-diabetic formulation enlisted in Ayurvedic lexicon. Sandhana Kalpana is mentioned under both Ahara and Aushadha Varga, and Asava-arishta were used as food supplements since Vedic period. Devdarvadyarishta was prepared as per conventional method and was subjected to various analytical parameters and nutritional parameters to develop a standard. Methods: Three samples of Devdarvadyarishta were prepared in accordance with Bhaishjya Ratnavali and compared with the market sample on various physico-chemical and phytochemical parameters, high-performance thin-layer chromatography (HPTLC) profiling as well as nutritional value. Results: Respective mean values of prepared samples and market sample were found to be pH – 3.86 and 3.80, specific gravity – 1.0953 and 1.0934, refractive index − 1.41943 and 1.41910, total solid content % w/w − 22.05 and 22.92, reducing sugars 18.18 and 18.50, nonreducing sugars 0.84 and 0.91 and total phenolic content – 0.180% and 0.185%, alkaloids – 0.037% and 0.042%, tannins – 0.11% and 0.10%, saponins − 0.082% and 0.080%, flavonoids – 0.011% and 0.013%, terpenoids − 0.046% and 0.055% and Glycosides – 0.11% and 0.17%. Nutritive value showed mean carbohydrate % of 21.45, fat − 0.21%, protein − 0.07%, and total energy being 86.82 K cal in prepared samples whereas market sample showed 22.97% carbohydrate, 0.25% fat, 0.10% protein, and 88.35 Kcal energy. HPTLC analysis showed two major peaks at Rf 0.87 and 0.90 with Chloroform: methanol solvent system. Conclusion: Physicochemical data for standard of Devdarvadyarishta was set in as a Quality specification for the same. Nutritional value of Devdarvadyarishta, i.e., 86.82 kcal energy supports the ancient views of Sandhana products being used as food adjuncts.

Keywords: Devdarvadyarishta, nutritive value, physico-chemical evaluation, standardization


How to cite this article:
Goyal C, Joshi N, Sharma KC. Physico-chemical standardization and nutritional assessment of Devdarvadyarishta. J Ayurveda 2022;16:126-33

How to cite this URL:
Goyal C, Joshi N, Sharma KC. Physico-chemical standardization and nutritional assessment of Devdarvadyarishta. J Ayurveda [serial online] 2022 [cited 2022 Aug 10];16:126-33. Available from: http://www.journayu.in/text.asp?2022/16/2/126/349760




  Introduction Top


Asava-arishta can be envisaged as the most refined type of dosage form on account of their superiority in having longer shelf life.[1] Sandhana Kalpana involves hydroalcoholic extraction of active compounds and is considered superior over tinctures for absorption in gut, former being digested partly only.[2] The primary fundamentals behind fermentation remained almost similar from ancient times to modern era with basic difference being noticed in raw drugs, type of equipment used, methodology adopted for preparation, sterilization, and standardization. To cope up with emerging needs of public health, pharmaceutical industries and market worldwide, standardization and optimization of Ayurvedic preparations are required. Several Asava-arishta has been described in Ayurvedic text but with indication in prameha are very less. One such formulation is Devdarvadyarishta prescribed in AFI part 1 with indication in Prameha. Considering its anti-diabetic indication, it becomes very essential to evaluate its margin of safety and to establish data for its standardization. Also, Sandhana Kalpana is having both medicinal and nutritive values, i.e., it comes under both Aushadha and Ahara Varga, respectively.[3],[4] As Ahara is said to be responsible for the anabolism of all Dhatus in the body by providing nutrition to them, Sandhana Kalpana may also be said to offer the same benefits being included under the Ahara Varga.[5] In the present research, the preparation process optimization with conventional technique of Devdarvadyarishta, its standardization by comparing with market formulation to establish biomarker by high-performance thin layer chromatography (HPTLC). As no standard parameters are available in API or anywhere else for Devdarvadyarishta, so market sample was considered as standard for this particular study. Asava-arishta are very well known for their nutritional value since Vedic period as these were used as food supplements. Also, Ayurvedic products are marketed globally as food supplements and not as medicine, so nutritional value evaluation has also been carried out.


  Materials and Methods Top


Plant materials and formulations

The herbs used in Devdarvadyarishta (three samples-DVA1, DVA2, DVA3) preparation were procured from local market in Haridwar after authentication by subject experts. The marketed formulation of Devdarvadyarishta (DVA4-Vaidya Ratnam Oushadhshala Pvt. Ltd) was purchased from Ayurvedic shop in Haridwar.

Preparation of devdarvadyarishta

The present study was conducted at Department of Rasa Shastra and Bhaishjya Kalpana, Rishikul Campus, Haridwar, Uttarakahand Ayurved University. The in-house Devdarvadyarishta was prepared as per Bhaishjya Ratnavali.[6] The composition is given in [Table 1] which was the same for all the three samples. Decoction was prepared after overnight soaking of drugs in required quantity of water which was further filtered with muslin cloth. Afterward honey, Prakshep dravyas (after powdering and sieving), Woodfordia fruticosa flowers were added and stirred properly. The mixture was kept in porcelain jar, sealed, and kept for fermentation. After completion of fermentation, it was filtered, kept for maturation and packed [Figure 1].
Table 1: Composition of different samples of Devdarvadyarishta

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Figure 1: Steps involved in pharmaceutical processing of Devdarvadyarishta

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Physico-chemical characterization-Analytical tests [Figure 2] were carried out at International Testing Centre, Panchkula (Haryana).
Figure 2: Analytical Testing Parameters of Devdarvadyarishta

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Organoleptic characters

Determination of organoleptic characteristics, namely odor, taste, and color of all the samples was carried out.

pH

The pH of the formulation was determined by using Digital pH meter.[7]

Specific gravity

The weight difference method was used for specific gravity determination. Ten mL of water (at 25°C) was taken into a clean, dry pycnometer and weighed. It was emptied, dried and further; 10 mL of each sample was filled in the same pycnometer and weighed at the same constant temperature. Specific gravity of the sample was determined by dividing the weight of the sample by the equal weight of water (at 25°C).[8]

Refractive index

The refractive index was conveniently measured using the Abbe refractometer at 25°C employing the wavelength of the D line of sodium (λ = 589.3 nm), after calibrating the apparatus against distilled water whose nD20 at 25°C was 1.3225.[9]

Determination of total solid content

The total solid means the residue obtained when the specific amount of the preparation is dried to constant weight under specified conditions. Transfer accurately 50 ml of sample in an evaporating dish and evaporate to a thick extract on water bath. Extract the residue with 4 quantities, each of 10 ml, of dehydrated ethanol with stirring and filter it. Combine the filtrates to another evaporating dish which have been dried to constant weight and evaporate nearly to dryness on water bath. Add accurately 1 g of diatomite, stir thoroughly and dry at 105°C for 3 h. Cool the dish in a desiccator for 30 min and weigh immediately. Deduct the weight of diatomite added, the weight of residue is noted as total solid content of the sample.[10]

Determination of alcohol content and test for presence of methanol

A gas chromatograph was used to determine the alcohol content of samples as well as to detect the presence of methanol.[11]

Determination of reducing sugars

Twenty ml of sample was neutralized with sodium hydroxide and was then evaporated to half volume on water bath to remove alcohol. After cooling 10 mL of 21.9 g zinc acetate, 3 mL glacial acetic acid followed by 10.6 g potassium ferrocyanide was added and volume was made up to 100 mL with distilled water. To the Fehling Solution (10 mL), burette solution was added drop wise. It was boiled till blue color. Further, two drops of methylene blue were added and titration was carried out till the appearance of brick red color.[12]

Determination of nonreducing sugars

Twenty mL of sample was mixed with equal amount of distilled water and further boiled for 30 min. The solution was cooled and adjusted to pH 7. The volume was further made up to100 mL by addition of distilled water. To the Fehling Solution (10 mL), solution from burette was added drop wise. It was heated to boiling till blue color appeared, over the hot plate. Then, two drops of methylene blue were added and the titration was carried out till the appearance of brick red color.[12]

Total phenolic content

The samples were prepared and equally diluted with water. Then, 0.25 mL Folin Ciocalteu reagent (1N) and1.25 mL sodium carbonate solution (20%) was mixed into it and vortexed. The mixture was kept in incubator at 45°C for 45 min and futher analyzed at 725 nm spectro-photometrically. The samples were prepared in triplicate for analysis and the mean value of absorbance was obtained. The standard solution of gallic acid was prepared similarly and the calibration curve was constructed. The total phenolic content of extracts was expressed in terms of gallic acid equivalent (mg of GA/g of extract).[13]

Qualitative analysis

Qualitative tests for alkaloids and glycosides were performed as per method adopted by Suman Kumar et al.[14] Tannins, flavonoids, and terpenoids by Edeoga et al.[15] and saponins estimation was done as prescribed by Kokate.[16]

Quantitative analysis

Alkaloids

Five g of the sample was weighed into a 250 ml beaker and 200 ml of 10% acetic acid in ethanol was added and covered and allowed to stand for 4 h. This was filtered concentrated on a water bath to one-quarter of the original volume. Concentrated ammonium hydroxide was added drop wise to the extract until the precipitation was complete. The whole solution was allowed to settle, precipitate was collected and washed with dilute ammonium hydroxide and then filtered. The residue is the alkaloid, which was dried and weighed.[15]

Tannins

Two g of sample was defatted with 25 ml petroleum ether for 12 h and the marc was boiled for 2 h with 300 ml of double-distilled water. It was cooled, diluted up to 500 ml, and filtered. 25 ml of this infusion in 2 liter porcelain dish was mixed with 20 ml Indigo solution and 750 ml double distilled water. Then, it was titrated with 0.1N Potassium permanganate solution, 1 ml at a time, until blue solution changes to green. Thereafter, drops wise solution was added until the solution becomes golden yellow. Similarly, the mixture of 20 ml Indigo solution and 750 ml of double-distilled water was titrated and the difference between two titrations in ml was calculated. Each ml of 0.1N potassium permanganate solution is equivalent to 0.004157 g of total tannins.[17]

Flavonoids

Ten g of the sample was extracted repeatedly with 100 ml of 80% aqueous methanol at room temperature. The whole solution was filtered through Whatman filter paper No 42 (125 mm). The filtrate was later transferred into a crucible and evaporated into dryness over a water bath and weighed to a constant weight.[15]

Saponins

Twenty g of sample was added with 100 cm3 of 20% aqueous ethanol was added and heated over a hot water bath for 4 h with continuous stirring at about 55°C. The mixture was filtered and the residue re-extracted with another 200 ml 20% ethanol. The combined extracts were reduced to 40 ml over water bath and the concentrate was mixed with 20 ml of diethyl ether with vigorous shaking. The aqueous layer was recovered while the ether layer was discarded. The purification process was repeated. Sixty ml of n-butanol was added. The combined n-butanol extracts were washed twice with 10 ml of 5% aqueous sodium chloride. The remaining solution was heated on a water bath. After evaporation, the samples were dried in the oven to a constant weight and the saponin content was calculated as percentage.[16]

Glycosides

One g of the sample was soaked in 10 ml of 70% alcohol for 2 h and then filtered. The extract obtained was then purified using lead acetate and Na2HPO4 solution before addition of freshly prepared Buljet's reagent (containing 95 ml aqueous picric acid + 5 ml 10% aqueous NaOH). The difference between the intensity of colors of the experimental and blank (distilled water and Buljet's reagent) samples gives the absorbance and is proportional to the concentration of the glycosides.[18]

Terpenoids

One g of sample was soaked in alcohol for 24 h and filtered. The filtrate was extracted with petroleum ether and ether extract was treated as total terpenoid.[19]

Heavy metal analysis

Heavy metals were determined by atomic absorption spectrophotometry (AAS)-vapor generation analysis.[20]

Test for aflatoxins

Aflatoxins content was determined by HPLC.[21]

Test for specific pathogens

Test for the presence of Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, and Salmonella species was conducted as per methods given in API.[22]

Microbial load

Total bacterial and fungal count was estimated as per API.[23]

Nutritive value

Determination of crude fat using soxhlet apparatus,[24] proteins by Lane and Eynon's Method,[25] carbohydrates by Kjeldahl Method,[26] and energy by calculation method was done.[27]

High performance thin layer chromatography analysis

0.5 g of Standard/Sample of Devdarvadyarishta was taken in a 250 ml flask and 25 ml ethanol was added to it and refluxed for 15 min. It was then cooled and filtered. Test and reference solution 10 ul in two different spots was applied on a precoated silica gel plate (5 cm × 10 cm) of uniform thickness which was then developed in solvent system Chloroform: Methanol (9:1) to a distance of 9.0 cm. Scanning was done at 254 nm and 366 nm and identification was done on the basis of Spot and Rf value.[28]


  Results Top


Organoleptic characters

All the samples were found similar in color (Reddish brown), taste (Sweet and Astringent), odor (alcoholic smell), and consistency (thick).

Physico-chemical parameters

Three prepared samples of Devdarvyadrishta have shown approximately identical results of various physico-chemical parameters as compared to the market sample [Table 2]. Respective mean values of prepared samples and market sample were found to be pH – 3.86 and 3.80, specific gravity − 1.0953 and 1.0934, refractive index – 1.41943 and 1.41910, total solid content % w/w – 22.05 and 22.92, reducing sugars 18.18 and 18.50, nonreducing sugars 0.84 and 0.91, and total phenolic content − 0.180% and 0.185%.
Table 2: Physico-chemical parameters

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Quantitative phytochemical parameters

Results revealed the presence of alkaloids – 0.037% and 0.042%, tannins – 0.11% and 0.10%, saponins − 0.082% and 0.080%, flavonoids − 0.011% and 0.013%, terpenoids − 0.046% and 0.055%, and glycosides – 0.11% and 0.17% [Table 3].
Table 3: Quantitative phytochemical parameters

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Test for heavy metals

All the four heavy metals were found to be in permissible limits [Table 4].
Table 4: Test for heavy metals

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Test for aflatoxins

None of the aflatoxins, namely B1, B2, G1, G2 was detected in any of the sample.

Test for specific pathogen

All the four pathogens were found absent in all the samples.

Total microbial load

No fungus was detected in the samples whereas total bacterial count was found to be in permissible limit.

Nutritive value/100 ml of Devdarvadyarishta

Nutritive value showed mean carbohydrate % of 21.45, fat − 0.21%, protein − 0.07%, and total energy being 86.82 K cal in prepared samples whereas market sample showed 22.97% carbohydrate, 0.25% fat, 0.10% protein, and 88.35 Kcal energy [Table 5].
Table 5: Nutritive value/100 ml of Devdarvadyarishta

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HPTLC analysis showed two major peaks at Rf 0.87 and 0.90 with Chloroform: Methanol solvent system.


  Discussion Top


The main aim of the analysis is to check the quality for obtaining desired therapeutic effect. Hence, it is necessary to control batch to batch variation, which is possible only through standardization protocols. That is why the samples were analyzed for their organoleptic properties, physicochemical studies, and phytochemical parameters [Table 2] and [Table 3]. Asava-arishta are very well known for their nutritional value since Vedic period as these were used as food supplements. Furthermore, Ayurvedic products are marketed globally as food supplements and not as medicine, so nutritional value was evaluated in the present study.

nDevdarvadyarishta being composed of 28 herbs excluding water and honey, and there is no available reference standard of the formulation, it was not possible to carry out HPTLC profiling using all individual constituent markers. Hence, HPTLC of the most suited product both pharmaceutically and analytically, i.e., DVA1 was done considering DVA4 as standard in this particular case as almost comparable results of market sample are found for various analytical parameters tested in the present study. HPTLC using Chloroform: Methanol (95:5) at 254 nm shows 4 peaks in sample at Rf 0.50, 0.65, 0.81, 0.84 and 3 peaks in standard at Rf 0.62, 0.80, 0.83 whereas at 366 nm, two identical major area peaks corresponding to Rf value 0.87 and 0.90 were found in both standard and sample as shown in [Figure 3], [Figure 4], [Figure 5].
Figure 3: HPTLC Chromatogram of DVA1 and DVA4 at 254nm

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Figure 4: HPTLC Chromatogram of DVA1 and DVA4 at 366nm

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Figure 5: HPTLC profiling of DVA1 and DVA4 in Chloroform and Methanol

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Heavy Metals analysis by AAS detected the presence of Lead in all the samples but within normal limits whereas Cd, As and Hg was not detected in any of the sample. Test for Aflatoxins by HPLC method showed complete absence of any of the aflatoxins in each sample. Microbiological tests revealed none of the Pathogenic bacteria, namely E. coli, S. aureus, Salmonella spp, P. aeruginosa to be present in any of the samples. Total Bacterial count was found to be within the permissible limits and no fungus was detected in all samples.

Results revealed the presence of active principles, namely glycosides, alkaloids, terpenoids, flavonoids, tannins, saponins in Devdarvadyarishta which are already known to possess antioxidant and antidiabetic effects. Due to the presence of all these functional groups, Devdarvadyarishta having good nutritional value, i.e., 86.82 kcal can be very well used in diabetic patients.


  Conclusion Top


The present research work was based on standardization and optimization of Devdarvadyarishta preparation. Physicochemical data and process for standard of Devdarvadyarishta were set in as a quality specification for the same. It was found to possess good nutritional value, i.e., 86.82 kcal energy supporting the ancient views of Sandhana products being used as food adjuncts. HPTLC analysis showing two major peaks at Rf 0.87 and 0.90 with Chloroform: Methanol solvent system can be considered as the marker for the same. It provides scope for further validation of established monograph of Devdarvadyarishta and evaluation of its safety and efficacy as antidiabetic formulation.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.





 
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    Figures

  [Figure 1], [Figure 2], [Figure 3], [Figure 4], [Figure 5]
 
 
    Tables

  [Table 1], [Table 2], [Table 3], [Table 4], [Table 5]



 

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