Pharmaceutico Analytical Study of Kanakbindvarishta prepared by two different...

[Full title: Pharmaceutico Analytical Study of Kanakbindvarishta prepared by two different methods]

ORIGINAL ARTICLE July-Aug 2019 Journal of Ayurveda and Integrated Medical Sciences | July - Aug 2019 | Vol. 4 | Issue 4 182 Pharmaceutico Analytical Study of Kanakbindvarishta prepared by two different methods Dr. Shital S. Yadav, 1 Dr. Arati Dubewar, 2 Dr. Ashwin Shete 3 1 Post Graduate Scholar, 2 Professor & HOD, 3 Assistant Professor, Dept. of Rasashastra & Bhaishajya Kalpana, Dr. D. Y. Patil College of Ayurved & Research Center, Pimpri, Pune, Maharastra, INDIA. I NTRODUCTION Sandhana Kalpana has been divided into two types Madhya Janak Sandhan (alcoholic fermentation) and Amlajanak Sandhan (acidic fermentation). In Acidic preparation there is no sweetening agent and Sandhan Pravartak Dravyas (fermentative agent) is used and in Alcoholic preparation sweetening agent (e.g. Jaggery, Honey and Sugar) and Sandhan Pravartak Dravyas (fermentative agent) is used, e.g. Dhataki Pushpa ( woodfordia fructicosa flower). Kanakbindvarishta [1] is an Ayurvedic formulation Address for correspondence: Dr. Shital S. Yadav Post Graduate Scholar, Dept. of Rasashastra & Bhaishajya Kalpana, Dr. D. Y. Patil College of Ayurved & Research Center, Pimpri, Pune, Maharastra, INDIA. E-mail: yadavshital 308@gmail.com Submission Date: 13/07/2019 Accepted Date: 15/08/2019 Access this article online Quick Response Code Website: www.jaims.in DOI: 10.21760/jaims.4.4.25 described in Charaka Samhita in the context of Kushtaroga . In the reference the ratio to prepare the decoction of Khadira and which sweetening agent should be used is not mentioned, so it is taken from Sharangdhara Samhita . Kanakbindvarishta is an alcoholic type of fermentation procedure. This formulation has longer shelf life, quick absorption, action and excellent therapeutic efficacy. It is mainly used in Kilasa Kushta , Asthama, Cough, Fistula and Diabetes. Kanakbindvarishta was prepared by two different methods. One is a classical preparation and another one is modified preparation. In classical preparation Dhataki Pushpa Kalka was used and in another second modified preparation instead of Dhataki Pushpa Kalka , Dhataki Pushpa Phanta was used, Prakshepa Dravyas was added after seven days, Sandhi Bandhana was done after 7 days. Raw material testing was carried out like LOD, Ash, pH, extractive value and TLC. Analytical testing of finished product was carried out as pH, specific gravity, refractive index, total solid, viscosity, reducing sugar, alcohol content and density. Need of this study A B S T R A C T Kanakbindvarishta is a self generated alcoholic preparation of Ayurveda. It is described in Charak Samhita in the context of Kushta Roga (skin disorders). Khadir (Acacia catechu) is a main ingredient in this formulation. It is used in condition like Kushta (mainly used in Kaphaja Kushta ), Asthma, Cough, Fistula and Diabetes. In present study Kanakbindvarishta was prepared by two different methods, Pharmaceutical study was done along with analytical study. Analytical test of finished product like pH, specific gravity, refractive index, total solid content, viscosity, density, reducing sugar and alcohol content were done, LOD%, total Ash, pH and extractive value (water extractive, alcohol extractive) of raw material were also done. Two different type of Kanakbindvarishta compared with pharmaceutical as well as analytical aspect. As a result, slight changes were observed in both the preparations. Key words: Kanak, Acacia Catechu, Arishta, Sandhana, Fermentation, Kushta.

Dr. Shital S. Yadav et.al. Pharmaceutico Analytical Study of Kanakbindvarishta. ISSN: 2456-3110 ORIGINAL ARTICLE July-Aug 2019 Journal of Ayurveda and Integrated Medical Sciences | July - Aug 2019 | Vol. 4 | Issue 4 183 was to compare both preparation analytically and pharmaceutically. A IM OF THE STUDY Pharmaceutico Analytical study of Kankbindwarishta prepared by two different methods. O BJECTIVE OF THE STUDY 1 Classical preparation of Kanakbindvarishta . Ref. Charaka Chikitsa ( Kushtarogadhikar ) 7/76-79 2 Modified preparation of Kanakbindvarishta . 3 Pharmaceutico analytical study of both the preparations. M ATERIALS AND M ETHODS 1. Materials Table 1: Details of ingredients of Kanakbindvarishta Dravya Type Drug name Latin name [7] Part used [7] Quantity Samp le A Samp le B Adhara Dravya Khadira Acacia Catecha Khadirsa ra, Twak 1280 gm 1280 gm Prakshepa Dravya (Adheya) Amalaki Emblica Officinalis Fruit 40 gm 40 gm Haritaki Terminalia Chebula Fruit 40 gm 40 gm Bibhitaki Terminalia Bellirica Fruit 40 gm 40 gm Sunthi Zinziber Officinalis Kanda 40 gm 40 gm Maricha Piper Nigrum Fruit 40 gm 40 gm Pippali Piper Longum Fruit 40 gm 40 gm Vidanga Embelia Ribes Fruit 120 gm 120 gm Haridra Curcuma Longa Fruit 120 gm 120 gm Musta Cyperus Rotundus Root 120 gm 120 gm Vasa Adhatoda Vasica Leaf 120 gm 120 gm Indrayav a Holarrhen a Antidycent rica Seeds 120 gm 120 gm Daruhari dra Berberis Aristata Twak 120 gm 120 gm Amruta Tinospora Cordifolia Kanda 120 gm 120 gm Fermenta tive agents Dhataki Woodfordi a Fructicosa Flower 50 gm 50 gm Sweeteni ng agents [2] Jaggery 2.5 Kg 2.5 Kg Honey 1.25 Kg 1.25 Kg Others Katak Beeja Strychnous potatorum seeds Seeds 5 gm 5 gm Picture 1: Raw drugs of Kanakbindvarishta 2. Methods ▪ Standardization of ingredients ▪ Method of preparation ▪ Analytical test of finished product 2.1 Standardization of ingredients Drug standardization was done in research laboratory of Dr. D. Y. Patil College of Ayurveda and Research

Dr. Shital S. Yadav et.al. Pharmaceutico Analytical Study of Kanakbindvarishta. ISSN: 2456-3110 ORIGINAL ARTICLE July-Aug 2019 Journal of Ayurveda and Integrated Medical Sciences | July - Aug 2019 | Vol. 4 | Issue 4 184 Center, Pimpri, Pune and their values were compared with API. Table 2: Standardization of ingredients Name of drug LOD% Total Ash% pH % Extractive Value TLC Alcohol % Water % Solvent System Alcohol - Rf Water -Rf Khadira [8] 6.50 2 5.25 6 12 Toluene : Ethylacetate, 9:1 0.76 0.08, 0.77 Amalaki [8] 10.15 0.082 3.44 54 57.49 Toluene : Ethylacetate, 9:1 0.018, 0.84, 0.66 0.43, 0.056 Haritaki [8] 6.2 5 3.78 41.2 58.6 Toluene : Acetone : Glacial Acetic Acid : Formic Acid, 3.5:5:1.5:0.5 0.67, 0.77 0.22,0.51, 0.77 Bibhitaki [8] 4.6 5 4.58 8 38 Chloroform : Acetic Acid : Water, 5:4.5:0.5 0.45, 0.52, 0.67 0.46, 0.6 Sunthi [8] 10.75 5.5 5.05 3.2 12.8 Toluene : Ethylacetate, 7:3 0.35, 0.47, 0.54 0.28, 0.35, 0.42, 0.48, 0.57 Maricha [10] 5.4 5 7.03 7.6 11.4 Toluene : Ethylacetate, 7:3 0.44, 0.51, 0.61 0.25, 0.36 Pippali [11] 6.8 5.92 5.86 28.64 29.92 Toluene : Ethylacetate, 9:1 0.17, 0.68, 0.86 0.19 Vidanga [8] 8.16 3.5 5.07 10.4 11.2 Toluene : Acetone : Acetic Acid, 9:1:0.5 0.22, 0.56 0.37 Haridra [8] 7 7.5 5.88 8.6 13 n-Hexane : Ethyl Acetate, 7:3 0.05, 0.14, 0.30, 0.41, 0.53, 0.58 0.1, 0.15, 0.31 Musta [10] 6.6 7 6.20 7.6 11.6 Toluene : Ethylacetate, 9:1 0.24, 0.58 0.38 Vasa [8] 4.6 17 5.64 5 27.2 Methanol : Sulphuric Acid, 5:5 0.33, 0.40, 0.78 Indrayava [10] 4.40 7.5 6.48 12.4 16.8 Chloroform : Methanol, 5:5 0.83 0.55 Daruharidra [9] 4.1 2 6.28 6.2 9.4 Toluene : Water : Glacial Acetic Acid, 6:3:1 0.22, 0.73 0.43

Dr. Shital S. Yadav et.al. Pharmaceutico Analytical Study of Kanakbindvarishta. ISSN: 2456-3110 ORIGINAL ARTICLE July-Aug 2019 Journal of Ayurveda and Integrated Medical Sciences | July - Aug 2019 | Vol. 4 | Issue 4 185 Amruta [8] 5.9 0.169 4.43 3.92 11.4 Toluene : Ethylacetate, 9:1 0.08, 0.83, 0.33, 0.66 0.28, 0.67 Jaggery 7.5 2.5 Honey 4.30 Glacial Acetic Acid: Chloroform : Ethanol, 2.1:2.3:1.5 0.39, 0.55, 0.94 Dhataki Pushpa 9.2 1.7289 4.65 8.4 11.54 Table 3: Sweetening Agents Honey [12] Parameter Test observation Color in Aqueous solution Light yellow pH 4.13 Specific gravity (g/ml) 1.41 g/ml Refractive index 1.4892 Viscosity 2928.74 cp TLC (Glacial Acetic Acid : Chloroform : Ethanol, 2.1:2.3:1.5) 0.39, 0.55, 0.94 Jaggery [12] Parameter Test observation Color in Aqueous solution Light yellow pH 4.13 Specific gravity (g/ml) 1.41 g/ml Refractive index 1.4892 Viscosity 2928.74 cp TLC (Glacial Acetic Acid : Chloroform : Ethanol, 2.1:2.3:1.5) 0.39, 0.55, 0.94 2.2 Method of preparation Following are the important elements in preparation of Kanakbindvarishta; 1 Adhara Dravya - Water, Decoction of Khadira Twak 2 Prakshepa Dravya ( Adhey ) - All Prakshepa Drvayas mentioned in above chart. 3 Sweetening agent - Honey and Jaggery 4 Fermentative agent - Dhataki Pushpa 2.2.1 Sample A [4][5] 1 Coarse Khadira bark was mixed with 16 parts of water and soaked overnight. 2 Decoction was prepared by classical method on mild flame. 3 Decoction was filtered through cloth and when it got cool jaggery and honey mixture was stirred till jaggery get dissolved. 4 Ceramic jar was sterilized by Dhoopana method. 5 The decoction was poured in sterilized ceramic jar and Prakshepa Dravyas with Dhataki Pushpa Kalka were added. 6 The container was sealed with mud smeared cloth and kept in clean and dry room for fermentation. 7 After one month fermentation was completed. 8 Sandhana Pariksha were carried out (Crackling sound and candle flame test) 9 Fermented material was filtered through cloth and was collected in sterilized jar and Kataka Beeja ( strychnous potatorum linn seeds) were added for Nirmalikarana. [5][6] 10 Analytical tests were carried out.

Dr. Shital S. Yadav et.al. Pharmaceutico Analytical Study of Kanakbindvarishta. ISSN: 2456-3110 ORIGINAL ARTICLE July-Aug 2019 Journal of Ayurveda and Integrated Medical Sciences | July - Aug 2019 | Vol. 4 | Issue 4 186 Picture 2: Preparation of Khadira Kwatha Picture 3: Filtration of Kwath a Picture 4: Addition of Prakshepa Dravyas Picture 5: Dhoopana of Jar Picture 6: Pouring the mixture in Jar Picture 7: Sandhi Bandhana

Dr. Shital S. Yadav et.al. Pharmaceutico Analytical Study of Kanakbindvarishta. ISSN: 2456-3110 ORIGINAL ARTICLE July-Aug 2019 Journal of Ayurveda and Integrated Medical Sciences | July - Aug 2019 | Vol. 4 | Issue 4 187 Picture 8: Galana of Sandhana 2.2.2 Sample B 1 All procedure, ingredient and their quantity were same as mentioned for sample A. 2 But instead of Dhataki Pushpa Kalka 200 ml Dhataki Pushpa Phanta were added and Prakshepa Dravyas were added after 7 days. Picture 9: Preparation of Dhataki Phanta 2.2.3 Arishta Siddhi Lakshana Following Lakshanas were found in sample A and B [5] ▪ The preparation had the characteristics as typical aromatic and alcoholic odor. ▪ There was no crackling sound heard in ceramic jar. ▪ Burning candle burns brightly when placed in just above Sandhana Patra . ▪ Powdered drugs settled down completely ▪ Arishta was clear, no froth was there on the top Picture 10: Candle Flame Test Picture 11: Clearity Test 2.3 Analytical test of finished product Instrument used for analysis of Arishta are as following. 1 pH meter 2 Abbe’s refractometer 3 Pyknometer 4 Oswald viscometer

Dr. Shital S. Yadav et.al. Pharmaceutico Analytical Study of Kanakbindvarishta. ISSN: 2456-3110 ORIGINAL ARTICLE July-Aug 2019 Journal of Ayurveda and Integrated Medical Sciences | July - Aug 2019 | Vol. 4 | Issue 4 188 5 Water bath 6 Weighing balance 1. pH The pH of solution provides a useful practical means for the indication of the acidity or alkalinity of a solution. The pH value of Kanakbindvarishta preparation was determined by using 1.021% w/v solution of potassium hydrogen phthalate as a primary standard and then pH of Kanakbindvarishta was determined. 2. Refractive Index The refractive index of formulation was found out by using Abbe’s refractometer 3. Viscosity The viscosity is the internal resistance to the flow of fluid. The viscosity of Kanakbindvarishta was determined by using Oswald viscometer. The Ostwald viscometer cleaned, dried and clamped in a vertical position. Both bulbs A and B was immersed in a constant temperature bath. Then taken specified volume of distilled water into the bulb A and sucked the liquid into the bulb B just above the mark M, about half of the bulb A still contain the liquid, the time of flow of the liquid level to fall from the mark M to the mark X was determined. A stopwatch was used to determine the time. 4. Specific gravity A specific gravity is the ratio of specific weight of the material to the specific weight of the distilled water. A specific gravity bottle of 10 ml capacity was cleaned, dried and weighed, it is filled upto the mark with water at the required temperature and weighed. The specific gravity bottle was next filled upto the mark with the sample. The specific gravity was determined by dividing weight of the sample expressed in grams by the weight of the water, expressed in grams. 5. Total solid content 25 ml of formulation was taken in evaporated dish which was previously weighted and allowed to evaporate so that only solid content remains in the dish and rest of the fluid gets evaporated, then it gain weight and the solid content of formulation calculated. Analytical test Sample A Sample B pH 4.27 4.31 Specific gravity 1.085 g/ml 1.1588 g/ml Refractive Index 1.3745 1.3789 Total Solid 74.36 gm 71.96 Viscosity 1.855 c.p. 1.8640 c.p. Density 1.1676 g/ml 1.1588 g/ml Reducing Sugar Present Present Alcohol Content 6.22 6.82 O BSERVATION AND R ESULTS Sample A and B of Kanakbindvarishta were subjected to organoleptic and physico-chemical studies in order to develop analytical profile. The following parameters were carried out in this phase. Parameters Sample A Sample B Rupa (Color) Light reddish Dark reddish Rasa (Taste) Tikta, Kashaya, Madhura Tikta, Kashaya, Madhura Gandha (Odor) Slightly Alcoholic Alcoholic Sparsha (liquid consistency) Thin Thin Determination of pH, specific gravity, refractive index, total solid content, viscosity, density, reducing sugar and alcohol content of sample A and B is as below.

Dr. Shital S. Yadav et.al. Pharmaceutico Analytical Study of Kanakbindvarishta. ISSN: 2456-3110 ORIGINAL ARTICLE July-Aug 2019 Journal of Ayurveda and Integrated Medical Sciences | July - Aug 2019 | Vol. 4 | Issue 4 189 Analytical test Sample A Sample B pH 4.27 4.31 Specific gravity 1.085 g/ml 1.1588 g/ml Refractive Index 1.3745 1.3789 Total Solid 74.36 gm 71.96 Viscosity 1.855 c.p. 1.8640 c.p. Density 1.1676 g/ml 1.1588 g/ml Reducing Sugar Present Present Alcohol Content 6.22 6.82 Fehling’s Test Benedict’s Test Finished Product DISCUSSION Kanakbindvarishta was prepared by two different methods. All precautions were carried out for making sample A and B. Standardization of raw drug was done carefully. Decoction was prepared as mentioned in classical test. Mixture was heated on mild flame as the quantity get reduced 1/4 th of initial volume. All used equipment was sterilized. Dhoopana of jar was carried out with Haridra , Jatamansi , Guggula , Sarjarasa and Vacha as it was necessary to clean jar properly to avoid growth of fungi which might be promoted due to organisms present in the water used to clean the vessel. The filtered Arishta was clear without froth at the top. The preparation had the characteristics as typical aromatic and alcoholic odor. It was stored in air tight jar. Dhataki Pushpa flower was used in both sample A and B preparation, the inoculum of yeast comes from Dhataki Pushpa ( woodfordia fructicosa flowers), which contains the wild species of the yeast, which is necessary for fermentation process [5] Fermentation is incomplete oxidation of sugar into ethanol and CO 2 in the absence of O 2 , brought about by enzymes , invertase and zymase secreted by yeast cells.

Dr. Shital S. Yadav et.al. Pharmaceutico Analytical Study of Kanakbindvarishta. ISSN: 2456-3110 ORIGINAL ARTICLE July-Aug 2019 Journal of Ayurveda and Integrated Medical Sciences | July - Aug 2019 | Vol. 4 | Issue 4 190 Honey and Jaggery were also used for fermentation, Jaggery contains sucrose, glucose and phosphate. Due to phosphate fermentation process increases. Honey contains nitrogenous elements which helps in fermentation. Jaggery has Saraka and Mutrala properties and honey has Raktadoshaghna , Vranashodhana and Vranaropana property, that’s why both are effective in Kushta. [5] Even after completing Sandhana in one month turbidity was there, so to remove turbidity 5 gm Kataka Beeja ( stychnous potatorum linn) in both sample was added for Nirmalikarana process. Sychnous potatorum linn contains natural polyelectrolytes which can be used as coagulants to clarify turbidity [6] Sandhana examination were carried out, there was no crackling sound and candle flame also didn’t go off [3] CONCLUSION Kanakbindvarishta prepared by two different methods. Sample A prepared by classical method and sample B prepared by modified method. In sample B Dhataki Pushpa Phanta was used and Prakshepa drug added after 7 days. And in sample A Dhataki Pushpa Kalka was used. After fermentation process finished product obtained of sample A was 4400 ml and of sample B it was 4500 ml. Odor of sample A was slightly alcoholic and of sample B was alcoholic. This study has future scope in antimicrobial and clinical study (Skin disorders). Analytical tests were carried out on both these two samples. There is difference in total solid content (residue). Total solid content of sample A was 74.36 gm and of sample B it was 71.96 gm. pH of sample A was 4.27 and of sample B it was 4.31. Refractive index of sample A was 1.3745 and of sample B was 1.3789. Viscosity of sample A was 1.855 c.p. and of sample B it was 1.8640 c.p. Density of sample A was 1.1676 g/ml and of sample B was 1.1588 g/ml. Reducing sugar was present in both sample. Alcohol content in sample A was 6.22 and in sample B it was 8.82. REFERENCES 1 Vd. Harish Chandra Singh Kushwaha, Charak Samhita Uttarardha, Edition 2009,Chaukhamba Orientalia,Varanasi, 2009, Pg. no. 206. 2 Dr. Brahmananda Tripathi, Sharangdhar Samhita, Edition 2015, Chaukhamba Surbharati Prakashan, Varanasi, Pg. no 164-165 3 Siddhinandan Mishra, Bhaishajya Kalpana, Edition 2004, Chaukhamba Surbharati Prakashan, Varanasi, Pg. no 249-256 4 Kaviraj Devisingh Vitthal, Bruhadasawarishta Sangraha, Edition 1990, Chaukhambha Sanskrut Pratishthan, Delhi, Pg. no 27 5 Vd. Gangadhar Vishnushastri Puranik, Dhootpapeshwar Prakashan, 2 nd Edition, Mumbai, Pg. no 186-229 6 N. Packialakshmi, C. Suganya, V. Guru, Studies on – Strychnous Potatorum seed and screening the water quality assessment of drinking water, 2014 IJRPKJ 3(5)380-396. 7 Dr. D. R. Lohar, Protocol for testing of Ayurved, Siddha, Unani medicines. Department of Ayush, Ministry of Health and Family welfare, Pg. no 49,50,54,124 8 The Ayurvedic Pharmacopia of India. Ministry of Health and Family welfare, Department of Ayush, Part-1, Vol – 1, Pg. no 9, 44, 55, 70, 77, 80, 122, 177, 207, 209 9 The Ayurvedic Pharmacopia of India. Ministry of Health and Family welfare, Department of Ayush, Part-1, Vol – 2, Pg. no 66, 10 The Ayurvedic Pharmacopia of India. Ministry of Health and Family welfare, Department of Ayush, Part-1, Vol – 3, Pg. no 105, 172, 192 11 The Ayurvedic Pharmacopia of India. Ministry of Health and Family welfare, Department of Ayush, Part-1, Vol – 5, Pg. no 51 12 The Ayurvedic Pharmacopia of India. Ministry of Health and Family welfare, Department of Ayush, Part-1, Vol – 6, CCXL ******************************* How to cite this article: Dr. Shital S. Yadav, Dr. Arati Dubewar, Dr. Ashwin Shete. Pharmaceutico Analytical Study of Kanakbindvarishta prepared by two different methods. J Ayurveda Integr Med Sci 2019;4:182-190. http://dx.doi.org/10.21760/jaims.4.4.25 Source of Support: Nil, Conflict of Interest: None declared.

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Arishta, Khadira, Saraka, Kalka, Phanta, Mutrala, Charakasamhita, Vranashodhana, Samdhibandhana, Jar, Vranaropana, Fermentation, Fermentation process, Decoction of khadira, Decoction, Specific gravity, Ayurvedic formulation, Turbidity, Refractive index, Prakshepa Dravya, Viscosity, Classical preparation, Dhataki Pushpa, Total solid content, Alcohol content, Ayurvedic Pharmacopia, Sharangdhara Samhita, Modified preparation, Reducing sugar, Kushtaroga, PH meter, Ethanol, Benedict's test, Fehling's test, Physico-chemical studies, Sweetening agent, Invertase, Analytical testing, Alcoholic fermentation, CO2, Organoleptic, Honey and Jaggery, Inoculum.