A A M J Anveshana Ayurveda Medical Journal

A A M J Anveshana Ayurveda Medical Journal www.aamj.in ISSN: 2395-4159 Research Article Pharmaceutico Analytical study of a herbomineral compound Yakritśūla Vināśini Vatika Anita Mali 1 Prakash Jondhale 2 Sanjay Kumar 3 K Shankar Rao 4 A b s t r a c t Yakritśūla Vināśini Vatika (YSV) is a herbo-mineral compound used in the management of Liver Diseases. Liver diseases are considered as fatal & life threatening. The formulation is mentioned in of the latest classics, Bhaishajyaratnavali. It is a combination of Navasādara and Saindhava lavana and 6 herbal ingredients with Putikambu swarasa (Chirabilva) levigation. Till date no standards are available for the above drug. Need of the hour is to revalidate the safety and efficacy of the above said formulation. Validation is defined as a procedure that demonstrates the ability of consistently producing a product with the established specifications under ideal conditions. The analytical profile of the drug by assessing its organoleptic, physico-chemical (including hardness, disintegration time, ph) as well as phytochemical parameters (flavonoids, tannins, alkaloids etc). Key words: Yakritśūla Vināśini Vatika, Herbomineral compound, Analytical profile, Antioxidant property. 1 Assistant Professor, Dept. of Agada Tantra & Vyavahara Ayurveda, 2 Assistant Professor, Dept. of Rasashastra & Bhaishajya Kalpana, AACH, Sirsa, Haryana. 3 Assistant Professor, 4 Professor & Head, Dept. of Rasashastra & Bhaishajya Kalpana, NIA, Jaipur. CORRESPONDING AUTHOR Dr. ANITA MALI Assistant Professor, Department of Agada Tantra & Vyavahara Ayurveda, AACH, Sirsa, Haryana. Email: dranita.mali@gmail.com AAMJ / Vol. 2 / Issue 3 / May June 2016

INTRODUCTION A ccording to the latest W.H.O. data published in April, 2011 death due to liver disease in India has reached 2.3% of total deaths. India stands 27th in the world. Liver disease is one of the major causes of morbidity and mortality affecting humans of all ages. Liver diseases are mainly caused by toxic chemicals, excess consumption of alcohol, viral infections, and drugs like paracetamol, antibiotics, and autoimmune disorders. Most of the hepatotoxic chemicals damage liver cells mainly by inducing lipid peroxidation and other oxidative damages. [i] In the present scenario where 80% of the world population has no access to the benefits of Western medicines due to financial constraints, it is quite necessary to emphasize the relevance of traditional remedies which constitute a major part of the health care system in the developing countries and are also entering the therapeutics in the developed countries. [ii] Till date, there is no effective medicine for hepatic disorders, such as hepatitis. Consequently, control of liver diseases has become a major goal of modern medicine. As it is the function of hepato-protective agents to interfere with these pathological processes by blocking their evolution and helping recovery, the development of new hepato-protective drugs is the need of the hour. The Ayurveda has a long tradition of treating liver diseases using herbal medicines. Herbal drugs play a major role in the management of various liver disorders most of which speed up the natural healing processes of the liver, have a combined action on the liver and the entire digestive system and also restore proper functioning of liver. [iii] Considering the above facts an attempt has been made to standardize the pharmaceutical preparation of Yakritśūla Vināśini Vatika. [iv] MATERIAL & METHODS Raw drugs were procured from NIA pharmacy, and pharmaceutical study was carried out at the Rasashastra and Bhaishajya Kalpana practical laboratory. Ingredients and their various proportions are mentioned in Table no.1. Pharmaceutical Processing Preparation of powders of crude drugs Śodhana of Navasādara Mixing of Powders Process of Bhāvana Formation of Vati Table No. 1. Ingredients of Yakritashula Vināśini Vatika S.N. Name Origin Latin Name Part used Pro-portion Quantity of Sample (gm) 1 Navasādara Mineral Ammonium chloride - 1 part 3 2 Saindhava lavana Mineral Rock salt halite / Chloride of sodium - 2 part 6 3 Chitraka Herbal Plumbago zeylanica Rt.bark 10 part 30 4 Kokilākśa Herbal Asteracantha longifolia Seed 10 part 30 5 Rohitak Herbal Tacoma undulate Bark 10 part 30 6 Yavāni Herbal Tachyspermum ammi Fruits 10 part 30 7 Putikambu (Swarasa) Liquid for levigation Holoptelia integrifolia Leaves. Q.S. Total Quantity. 129 gm S. No Drug Name Part Used Table No. 2: Observation during churna nirmana. Wt. of Raw Drugs Wt. of Powdered drugs Color of Powdered Loss (gm) 1. Saindhava - 100 98 White 2 2.04 2. Kokilākśa Seed 98 95 Brown 3 3.15 3. Rohitak Bark 98 92 Light Brick-red 6 6.52 4. Yavāni Fruit 95 89 Light yellow 6 6.74 5. Chitraka Rt.bark 97 92 Brown 5 5.43 % Loss AAMJ / Vol. 2 / Issue 3 / May June 2016 657

S. N. Drug Initial quantity Anita et.al., : Pharmaceutico Analytical study of a herbomineral compound Yakritśūla Vināśini Vatika Table No. 3: Observation during Navasādara Śodhana [v] Time required for śodhana Yield after Śodhana (gm) % Yield Loss in Śodhana process (gm) 1. Navasādara 150 gm 5 hrs 129 gm 86 % 21 2. Navasādara 150 gm 4hrs 55 min. 131.2gm 87.46% 18.8 3. Navasādara 150 gm 5hrs 20 min. 130.8gm 87.2% 19.2 Mean 150 5hrs 5 min. 130.3 86.88% 19.6 Table No. 4. Show Organo-leptic Characters of three samples of Navasādara. Table No. 6: Organoleptic characters of swarasa Navasādara Organoleptic Characters Colour Touch Odour Taste Greyish- White White Crystal Powder Before śodhana Odourless Corrosive After śodhana Odourless Corrosive Swarasa Taste Appearance Odour Cirbilva Katu (Bitter) Liquid Typical to Cirbilva Table No.5: Showing drugs using for Bhāvana in YSV. Swarasa Colour of Swarasa Material consumed Total Swarasa obtained Time in extraction process Yield (in %) Cirbilva Dark green 3.5 kg 975ml 45 min. 27.85 Table No. 7: Showing Seven Bhāvana of YSV. S. N. Drug Used Part No. of Bhāvana Quantity of Swarasa consumed for each Bhāvana Colour of Mixture after each Bhāvana of Swarasa Increase in Weight (gm) 1. I 220 ml Brown 168 2. II 190 ml Brown 162 3. III rd 160 ml Brown 160 Leaves Cirbilva 4. swarasa IV th 130 ml Brown 155 5. V th 120 ml Brown 147 6. VI th 80 ml Brown 148 7. VII th 75 ml Brown 150 Total quantity 975 ml PROCEDURE All the above mentioned ingredients (from no.1 to no.6) were taken as per mentioned quantity and mixed together homogeneously in khalva yantra and sufficient quantity of Cirbilva patra swarasa added to it and then triturated with it for 4 hrs/ day, till it get dry sufficient. Again sufficient quantity of Cirbilva patra swarasa was added and triturated till it gets dry completely. Once the sufficient consistency was obtained, pills were prepared from that mass. The Experiment was repeated with other two samples and average values were taken. AAMJ / Vol. 2 / Issue 3 / May June 2016 658

Table No.8. Showing mean of the YSV samples, during pharmaceutical procedure. S.N. Parameters Sample 1 Sample 2 Sample 3 Mean 1 Raw drug quantity 129 gm 129 gm 129 gm 129 gm 2 Equipments: Mortar Boat shaped-pashana Round shaped- Porselin Boat shaped-pashana L= 38 cm Dia.=16 cm L= 30 cm B=25.5 cm - B=22.5 cm H=15 cm H=7.5 cm H=14 cm D=8.1 cm D=6.3 cm D=7.5 cm Pestle L=23.2 cm L=14.2 cm L=21.6 cm 3 Total material consumed 3.5 kg 3.5 kg 3.5 kg 3.5 kg 4 Total swarasa 975 ml 989 ml 982 ml 982 ml 5 Bhāvana 7 7 7 7 6 Total Duration of trituration 7 day 7 day 7 day 7 day 7 Time in extraction process 45 min 48 min 50 min ~47 min 8 % yield 27.8% 28.25% 28.05% 28.03% 9 10 Weight of mass after completion of Mardana Weight of powder after completion of Mardana 178 gm 179 gm 176 gm 177.66 gm 150 gm 152 gm 151.5 gm 151.16 gm 11 Final wt. of pills 148 gm 150 gm 149.5 gm 149.16 gm 12 Colour Dark brown Dark brown Dark brown 13 Odour Typical to Ajwain Typical to Ajwain Typical to Ajwain 14 Taste 15 Precautions Tikta, Kashaya lavana Dry cotton cloth was used during swarasa extraction. Tikta, Kashaya lavana Wet cloth was used Tikta, Kashaya lavana Wet cloth was used - - Analytical study Physico-chemical constants like LOD, Acid Insoluble ash, Extraction values, Hardness of Pills, Disintegration Time, Fineness of the Particles, TLC and AAS value were carried out along with screening for phyto-chemical constituents and microbial contamination. A. Organoleptic Characters B. Physico- Chemical Parameters C. Extraction Procedure D. Thin Layer Chromatography (TLC) E. Microbial Count F. AAS value of YSV RESULTS Results of analytical studies carried out for the formulation are depicted in the following tables (Tables 9 to 15) Table No.9: The Organo-leptic characters of the sample is tabulated as below- Sample Colour Odour Taste Touch Shape Churna Brown Typical to ajwain, Madhura, Lavana, Tikta, Kashaya Mild rough Powder form Vati Brown Typical to ajwain,, Madhura, Lavana, Tikta, Kashaya, Soft, smooth Spherical AAMJ / Vol. 2 / Issue 3 / May June 2016 659

Table No.10: Showing the physicochemical analysis of YSV samples. S. No. Parameters Sample Value 1 ph Value [vi] (10% w/v )in water Vati (powder) 5.19 2 Moisture content [vii] (%w/w) / LOD Vati (powder) 8.338 3 Total Ash [viii] (%w/w) Vati (powder) 12.193 4 Acid Insoluble Ash [ix] (%w/w) Vati (powder) 0.387 5 Alcohol soluble extractive [x] (%w/w) Vati (powder) 22.145 6 Water soluble extractive [xi] (%w/w) Vati (powder) 32.738 7 Average weight Vati 0.46170 gm 8 Hardness of Pills Vati 6-7.5 kg/cm2 9 Disintegration Time [xii] Vati 2hr.45 min. to 2hr.50 min. 10 Fineness of the Particles. YSV powder All the particles pass through sieve no. 100. EXTRACTION PROCEDURE For TLC profile & extractive values of YSV were Soxhelet extracted successively in Petroleum ether, Benzene, Chloroform, Ethyl acetate, Methanol and Distilled water for 6 h. These extracts were filtered, evaporated to dryness and weighed. Each extract (10 mg) was dissolved in 10 ml to make a concentration of 1mg/ml used for further studies. Table No.11. : Extractive values of YSV S.N. Extracts Yield (% w/w) 1. Petroleum Ether 3.4 2. Benzene 3.54 3. Chloroform 3.27 4. Methanol 7.5 5. Aqueous 10.6 THIN LAYER CHROMATOGRAPHY (TLC) (13) YSV - Stationary phase : Silica gel 60F 254 Solvent system: Petroleum ether extracts - Pet ether : Benzene (1:3), Benzene extracts - Benzene : Ethyl acetate (1:2), Chloroform extracts - Chloroform: Methanol (1:2), Ethyl acetate extracts - Benzene : Ethyl acetate (2:1), Methanol extracts - Chloroform: Methanol (2:1). Solution apply : 10 microlitres (for Methanol extract) Sprayer : Vanillin-Sulphuric acid mixture. R f values (on visible light) Table No.12. : Rf values of samples (on visible light): YSV S.No. Extract Rf Value 1. Pet. Ether 0.7 2. Benzene 0.57, 0.32 3. Chloroform 0.45, 0.60 4. Ethyl acetate 0.36 5. Methanol 0.39, 0.27,0.23 6. Aqueous 0.69 Table No.13: Showing Microbial Count: (14) S.No. Microbes Sample Values 1. Escherichia coli YSV powder Absent/gm 2. Salmonella - Absent/gm 3. Pseudomonas aeruginosa 4. Staphylococcus aureus - Absent/gm - Absent/gm Table No.14: Showing: AAS value [xv] of YSV. S.N. Heavy metal Values in ppm 1. Arsenic 2.740 3.0 2. Lead 8.83 10.0 3. Mercury Not detected 1.0 Permissible limits in ppm (WHO) PHYTOCHEMICAL TEST Table No.15: Showing Qualitative Phytochemical tests for extracts of YSV. S.N. Tests Pet. Ether Benzene Chloroform Ethyl- acetate Methanol Water 1. Terpenoids ++ ++ ++ -ve ++ ++ 2. Tannins -ve -ve + + ++ ++ 3. Saponins -ve + -ve ++ + ++ 4. Flavenoids -ve -ve -ve ++ ++ ++ 5. Alkaloids -ve + + + ++ ++ 6. Glycosides -ve + + + ++ ++ AAMJ / Vol. 2 / Issue 3 / May June 2016 660

DISCUSSION In pharmaceutical process (Table no.1 to7) the powder mix (129g) was levigated by adding the juice and triturated for 4 hours. The process was repeated for 7 consecutive days and after 7 th cycles when the mass became homogenous the pills were prepared by hand moulding. In this experiment it was observed that the liquid added at each time was absorbed by the kokilākśa and become very mucilaginous material. It was observed for initial 3 days and after that the material becomes soft and homogenous mass, because of kokilākśa more liquid was consumed. The amount of extract consumed was 982 ml. The prepared pills were dried in shade and packed in air tight bottles. The pills are of brown in colour. It took overall 8 days and the weight of the mass obtained before making the pills was 177.66 gm which may be because of the addition of contents from liquid. After shade drying the weight of the pills observed was 149 gm increased by 18gm - addition of solid contents and the loss of moisture content are the reasons for this. The making of pill process was repeated for three times. The results were showed in Table No.7. The colour of the pills was brown. The herbal drugs will give the same brown colour a natural phenomenon. The odour is typical to thymol like and it might be because of the ingredient ajwain in which thymol is one of the active constituent. The taste is madhura, lavana and bitter. The saline taste is due to presence of salt and Navasādara. The bitterness might be due to ajwain. The ph value of the pills is 5.19% w/v. The reason might be presence of Navasādara and the herbal ingredients. Loss on drying was 8% w/w and this indicates the presence of moisture content in the pills and also to a little extent to the volatiles present in the formulation. The total ash content was 12% w/w and this is the indicative of the inorganic material present in the formulation. The herbal ingredients and to a little extent the minerals contribute to this. Acid insoluble ash is 0.387 % w/w. This is because of the presence of silica and the readings are very low and it means the silica matter is negligible in the preparation.the extractive values are the indicators of the extractable active ingredients present in the preparation. The readings show that a fair amount of active ingredients are present in the preparation. The average weight of the pill was 0.461mg. The pills were made by hand moulding and therefore there are likely chances for weight variation. The disintegration time taken was too long and beyond the standards. It might be because of the presence of the kokilākśa a mucilaginous material might not allowed to disintegrate easily. The TLC showed varied findings. The spots observed in different solvents showed in Table No.12.The study findings of microbial count shows that the material was sterile as no microbes were grown. The heavy metal content findings were also within the normal range. Phytochemical screening of YSV has revealed that the presence of flavonoids, glycosides, alkaloids, tannins & terpenoids, saponins. Phytoconstitutes like the flavonoids (Bohnert & Jensen, 1996), terpenoids, saponins & alkaloids are known to possess hepatoprotective activity. CONCLUSION The present effort to develop on SOP for the preparation of Yakritśūla Vināśini Vatika as well as to develop its analytical profile serve as a preliminary step towards standardization of the formulation. The phyto-chemical findings indicate the presence of various plant constituents like flavonoids, alkaloids, tannins etc. The presence of flavonoids etc. in extract may be responsible for its antioxidant and thus hepatoprotective activity. ΛΛΛΛ AAMJ / Vol. 2 / Issue 3 / May June 2016 661

REFERENCES i. Gupta LM & Raina R. Side effbcts of some medicinal plants. Curr. Sci 1998,75, 897-900. ii. Chattopadhyay MK. Herbal medicines. Curr Sci 1998,71, pg.5. iii. Shanmugasundaran P, Venkataraman S.epatoprotective and antioxidant effects of Hygrophila auriculata (K. Schum) Heine Acanthaceae root extract. J Ethnapharmacol 2006;104:124-8.4. API,Part-2nd, Vol-1st;2.2.4,pg. 234. iv. AFI,part 1st,vol.1st A-Formulation group;12/24. v. Sharma Sadananda,Rasatarangini,Motilal Banarasidas, New Delhi,11 edition, reprint 2009, 14/3-4. vi. Pharmacopia of Inadia (IP)-2010;2.2.24. vii. Ayuevedic Pharmacopia of Inadia, Part -2nd,Vol. -2st; 2.2.9, pg. 235. viii. Ayuevedic Pharmacopia of Inadia,Part-2nd, Vol.-1, 2.2.3; pg. 234. ix. Ayuevedic Pharmacopia of Inadia,Part-2nd, Vol-1st;2.2.4,pg. 234. x. Ayuevedic Pharmacopia of Inadia,Part-2nd, Vol-1st;2.2.6,pg. 235. xi. Ayuevedic Pharmacopia of Inadia,Part-2nd, Vol-1st;2.2.7,pg. 235. xii. Pharmacopia of Inadia (IP)-2010, 2.5.1; pg.187. xiii. Ayuevedic Pharmacopia of Inadia,Part-2nd, Vol- 1st;2.2.12, pg. 239. xiv. Pharmacopia of Inadia (IP)- 2010,2.2.9, pg. 40-45. xv. Pharmacopia of Inadia (IP)-2010,2.4.2 ; pg.109. Source of Support: Nil. Conflict of Interest: None declared How to cite this article: Anita et.al., : Pharmaceutico Analytical study of a herbomineral compound Yakritśūla Vināśini Vatika AAMJ 2016; 3:656 662 ΛΛΛΛ AAMJ / Vol. 2 / Issue 3 / May June 2016 662