Hetal P. Baraiya. et al. / International Journal of Biological & Pharmaceutical Research. 2015; 6(9):

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1 734 e- ISSN Print ISSN International Journal of Biological & Pharmaceutical Research Journal homepage: IJBPR PHARMACOGNOSTICAL AND PHYTOCHEMICAL STANDARDISATION OF NIMBADI GHANAVATI - AN AYURVEDIC POLYHERBOMINERAL FORMULATION Hetal P. Baraiya *, Harisha CR, Shukla VJ, Shilpa B Donga, L.P.Dei Department of Streeroga and Prasootitantra, I.P.G.T. and R.A., Gujarat Ayurved University, Gujarat, Jamnagar , India. ABSTRACT Nimbadi Yoga has been mentioned as Anubhuta Yoga by Vd. Devisharan Garga Ayurveda Upadhyaya in Nari Roganak is selected for the management of Swetapradara i.e. abnormal vaginal discharge. In present study Ghanavati Kalpana is selected as an oral formulation of this Yoga as dose maintenance is to be perfect in Vati form. Pharmacognostical, physicochemical and phyto-chemical (including High Performance Thin Layer Chromatography) observations revealed the specific characters of all active constituents used in the preparation. The present work was carried out to standardize the finished product of Nimbadi Ghanavati to confirm its identity, quality and purity. The presence of stone cells, oil globules, prismatic crystal, scleroid, starch grain, tannin content, trichome etc. were the characteristic features observed in the microscopy of the prepared drug. Identified phytochemical components (Carbohydrates, tannins, fanolic compound, glycosides, steroids and flavonoids etc.) support the intended action of the formulation in abnormal vaginal discharge. Nimbadi Ghanavati, a polyherbal formulation of six ingredients was prepared and HPTLC finger print profile was developed and it can be considered pharmacopial standard. Key Words: Abnormal vaginal discharge, HPTLC, Nimbadi Ghanavati, Pharmacognosy, Phytochemistry, Shwetapradara. INTRODUCTION In this present era of materialism, women are surrounded with various kinds of disease in which abnormal vaginal discharge is most common. It very much depends upon woman's own perception, power of observation and tolerance etc. or has great individual variation. Vaginal discharge is indicative of some white creamy, yellowish, greenish discharge from vagina as abnormal or pathological process in the genital tract. It usually shows symptoms in association with other illness and commonly caused by fungal, parasitic, bacterial and sexually transmitted disease like trichomonas vaginalis. In Ayurvedic texts, though various conditions are described in which Swetasrava occurs but the Swetapradara as a Corresponding Author Hetal P. Baraiya het.baria@gmail.com special disease is not mentioned anywhere in Brihatrayee and Laghutrayee. For bleeding per vagina Raktapradara, Pradara and Asrigdara and for white discharge Swetasrava or Yonisrava, Slesmala Yoni words have been used. The signs and symptoms of Slesmala Yoni Vyapada can be correlated with the term Swetapradara which is commonly seen in practice (Agnivesha PT. Brahmashankar Mishra, 2009). Commentator Cakrapani, Sharngadhara Samhita, Bhavaprakasa and Yogaratnakara have used the word Swetapradara for white vaginal discharge. Commentator Cakrapani (11 th centaury) has well narrated this term (Agnivesha, Vaidya Jadavji Trikamji Acharya, 2011). In Acharana Yoni there is no direct discription of vaginal discharge in classics, but due to uncleanliness and presence of microbes vaginal discharge along with severe itching is seen (Agnivesha PT. Brahmashankar Mishra, 2009). According to WHO -every year 333 million new

2 735 cases of curable vulvovaginitis infections are registered. 10% to 41% of women have had abnormal vaginal discharge at least once in a life. In the last 20 years incidence is doubled. So it is great scope of research to find out safe, potent, cost effective remedy from Ayurveda for the management of Swetapradara. Thus Nimbadi Ghanavati has been selected for the trial which has been mentioned as Nimbadi Yoga- Anubhuta Yoga by Vd. Devisharan Garga Ayurveda Upadhyaya in Nari Rogana (Devisharan Garga Ayurvedapadhyay, 1934). The selected compound in the present study i.e. each ingredient of Nimbadi Ghanavati is very easily available in sufficient quantity, non controversial and also economical. It also has properties like Tridoshashamaka especially Kapha Vata Shamaka, Krimighna, Vranashodhana, Vranaropana, Putihara, Kandughna as well as antimicrobial, antifungal effect which leads to Samprapti Vighatana & also prevention of recurrence (Anonymous 1, 2000). MATERIALS AND METHODS Collection of Raw Drugs Most of the raw drugs of Nimbadi Ghanavati were collected from Pharmacy of Gujarat Ayurved University, Jamnagar. Fresh Nimba patra had been collected from Botanical garden, Gujarat Ayurved University, Jamnagar Preparation of Nimbadi Ghanavati Nimbadi Ghanavati was prepared in Pharmacy of Gujarat Ayurved University, Jamnagar. Ingredients, part used and their required quantity for each Nimbadi Ghanavati of 500mg are given in Table-1. Pharmacognostical Evaluation As per API drugs which are used in the finished product of Nimbadi Ghanavati were identified and authenticated by the Pharmacognosy department. The identification was carried out based on the morphological, organoleptic features and microscopy of the prepared drug (Anonymous 2, 2006), (Anonymous 3, 2006).. Pharmaceutical Evaluation Physico-chemical Parameters Following parameters were analyzed for different physico-chemical parameters by today s routine methods at the pharmaceutical chemistry lab, IPGT& RA, Jamnagar (Anonymous 4, 2006). Physico-chemical Parameters for Nimbadi Ghanavati Uniformity of weight Loss on drying Total ash Methanol soluble extract PH Qualitative test for various functional groups The techniques employed to isolate active substance are termed as extraction method. Crude extracts obtained from such processes can be qualitatively tested to ascertain the presence of different types of components. Qualitative tests are used to detect the presence of functional groups, which plays very important role in the expression of biological activity (Weins C. and Hauck H., 1996), (Kokate, 1996). High Performance Thin layer Chromatography Measured volume of the sample was evaporated to dryness, the residue was extracted with methanol by maceration and the sample for spotting was prepared. HPTLC study of the samples was carried out (Kalasz H., Bathori M., 1996). RESULTS AND DISCUSSION Pharmacognostical study The initial purpose of the study was to confirm the authenticity of the drugs used in the preparation of Nimbadi Ghanavati. For that Finished product microscopy of the drugs was studied microscopically and microscopic characters of them were drowned. Results matched with the API and thus confirmed the genuineness of all the drugs used in the finished product. Organoleptic findings Nimbadi Ghanavati shows black in colour, Kashaya taste, slightly aromatic odour and hard to touch. Finished product microscopy Diagnostic characters of Ghanavati showed brown content of Haritaki, oil globule of Nimba, prismatic crystal of Nimba, rhomboidal crystal of Nimba, rosette crystal of Bibhitaki, scleroids of Haritaki, scleroid of Bibhitaki, scleroids of Amalaki, silica deposition of Amalaki, starch grain with hilum of Haritaki, starch grains of Bibhitaki, stone cells of Bibhitaki, stone cells of Haritaki, tannin content of Bibhitaki, trichome of Bibhitaki (Plate-1). Pharmaceutical Evaluation Physico-Chemical parameters of Nimbadi Ghanavati like ph, Loss on drying, water soluble extract, uniformity of weight and methanol soluble extract all were found to be within the normal range. Details are given in Table-2. Qualitative test for various functional groups reveals the presence of Carbohydrates, tannins, fanolic compound, glycosides, steroids and flavonoids in drug. Details are given in Table -3. HPTLC profile of methanolic extract of Nimbadi Ghanavati was done and details of number of spots and Rf value are given in Table-4 (Plate- 2).

3 736 Table 1. Ingredients of Nimbadi Ghanavati No. Drug Latin name Part used Ratio 1. Nimba Patra Azadirecta indica A. Juss. Fresh Leaf 4 part Triphala Dried Fruit 1. Haritaki Terminalia chebula Retz. Kwatha 2. 1 part 2. Bibhitaki Terminalia belerica Rox drugs 3. Amalaki Emblika officinalis Linn. Prakshepa 3. Madhu - 1/5 part 4. Su.Sphatika Al(so 4 )24H 2 O Powder 1/10 part Table 2. Physico-chemical parameters of Nimbadi Ghanavati Parameters Nimbadi Ghanavati 1 Uniformity of weight (gm) Maximum weight gm Minimum weight gm Average weight gm 2 Loss on drying (%) 84.91% 3 Ash value (%) 7.332% 4 Methanol soluble extract 35.73% 5 ph Value (By ph meter) 3.0 Table 3. Showing qualitative test for various functional group of Nimbadi Ghanavati S. No Component Test Result 1. Carbohydrates Molish Test +ve 2. Sugar Fehling s test - ve 3. Proteins T.S.+4%NaOH+CuSo4 - ve 4. Aminoacid Ninhydrin - ve 5. Steroids Salkowski +ve 6. Glycosides Keeler-Kiliani Test +ve 7. Saponin Foam test - ve 8. Flavonoids Lead acetate + ve 9. Alkaloids Dragendroff s test - ve 10. Tannins and Phenolic Compound Lead acetate +ve Table 4. High Performance Thin Layer Chromatography results for methanolic extract of Nimbadi Ghanavati Visualisation condition UV-366nm UV-254nm No. of Spot Rf Value No. of Spot Rf Value Fig 01. Brown content of Haritaki Fig 02. Oil globule of Nimba

4 737 Fig 03. Prismatic crystal of Nimba Fig 04. Rhomboidal crystal of Nimba Fig 05. Rosette crystal of Bibhitaki Fig 06. Sceroids of Haritaki Fig 07. Scleroid of Bibhitaki Fig 08. Scleroid of Haritaki Fig 09. Scleroids of Amalaki Fig 10. Silica deposition of Amalaki Fig 11. Starch grain with hilum of Haritaki Fig 12. Starch grains of Bibhitaki Fig 13. Stone cells of Bibhitaki Fig 14. Stone cells of Haritaki

5 738 Fig 15. Tannin content of Bibhitaki Fig 16. Trichom of Bibhitaki Fig 17. Showing HPTLC profile of Nimbadi Ghanavati Peak display 254 nm Peak display 366 nm CONCLUSION Pharmacognostical findings confirm the ingredients present in the finished product of Nimbadi Ghanavati and there is no major change in the microscopic structure of the drug during the pharmaceutical processes of preparation of Kwatha for Ghanavati. Identified phytochemical components like Tannins, steroids etc. are astringents and anti-inflammatory, thus prevent the inflammation and reduce the secretion of discharge, pain or tenderness, redness, swelling of vulva and vagina. It is inferred that the formulation meets minimum qualitative standards as prescribed by API at preliminary level. The results of this study may be used as the reference standard in further research undertakings of its kind. REFERENCES Agnivesha PT. Brahmashankar Mishra. Charaka Samhita with Vidyotini vyakhya. ed. Chaukhambha, Varanasi. Chikitsa sthana. 2009; 30(14): 842. Agnivesha PT. Brahmashankar Mishra. Charaka Samhita with Vidyotini vyakhya. ed. Chaukhambha, Varanasi. Chikitsa sthana. 2009; 30(18): 842. Agnivesha, Vaidya Jadavji Trikamji Acharya, Chakrapani. Charaka Samhita with Ayurvedadeepikavyakhya. ed. Chaukhambha, Varanasi. Chikitsa sthana. 2011; 30: 116. Anonymous 1, Govt. of India. Dept. of ISM&H, Ministry of Health & Family Welfare. Database on Medicinal plant used in Ayurveda C.C.R.A.S. New Delhi. 2000; 3: Anonymous 2, Govt. of India. The Ayurvedic Pharmacopeia of India. First edition, Part 1, New Delhi. 2006; 1(7): 8, 33, 62. Anonymous 3, Govt. of India. The Ayurvedic Pharmacopeia of India. First edition, Part 1, New Delhi. 2006; 2: 131. Anonymous 4, Govt. of India. The Ayurvedic Pharmacopoeia of India. 1st edition, part II, The Controller of publications, New Delhi. 2006; 3(03): 212. Berger GB. Flavour and fragrances chemistry, Bio-processing and sustainability, FB Chemie, institute, Hannover, Germany Devisharan Garga Ayurvedapadhyay. Anubhut. Striyo ki sampurn vishesha vyadhiyo ka vistruta sachitra krambaddha sahitya, anubhut purva chikitsa vidhi, evam sahaja prayog sangrah. 1934; 2: 3. Kalasz H and Bathori M. Present status and future perspectives of thin layer chromatography. LC-GC Int. 1996; 10: Kokate. Practical pharmacognosy. Nirali publication, Pune. 1996: Weins C and Hauck H. Advances and Developments in Thin Layer Chromatography. LC-GC Int., 1996; 9: