STANDARDIZATION OF THE HYDRO-ALCOHOLIC EXTRACT OF AYURVEDIC VAGINAL FORMULATION (NA) BY USING CHROMATOGRAPHY (HPTLC, HPLC) AND SPECTROSCOPY (UV-VIS & FTIR & GC-MS) METHODS

The research herbal formulation prepared by mixing dried stem barks of Azadirachta indica A. Juss. and Saraca asoca Roxb. in equal amounts is standardized through pharmacognostic and phytochemical studies for assessing its efficacy in the treatment of leucorrhoea/vaginitis/ excessive white discharge. Pharmacognostical analysis revealed total ash value of 9.43% having 1.89% acid insoluble ash, 4.62 pH and 7.70% moisture content and presence of nitrogen and halogens. Flavonoids, tannins, carbohydrates, glycosides, saponins and fixed oils were found present in the hydro-alcoholic extracts. The results also showed a high concentration of flavonoids compounds (43.37 µg quercetin equivalent/mg) and high phenolic content (101.22 µg gallic acid equivalent/mg). UV-Visible spectroscopy scanning showed a prominent peak at 277 nm in the hydro-alcoholic extract. HPTLC analysis at 254 nm, 277 nm, and 330 nm indicated the presence of tannic acid, gallic acid, quercetin and catechol. Similarly, HPLC analysis at 277 nm suggested the presence of gallic acid, tannic acid and catechol. FTIR analysis exhibited the presence of N-H stretching, C-H stretching, N-H bending, N-O asymmetric stretching, C-C stretching, C-H rocking, C-N stretching, C-O stretching, C-H wagging, C-N stretching, C-Cl stretching, C-H bending and C-Br stretching functional groups suggesting the presence of amines/amides, alkenes, nitro group, aromatics, alcohol/ carboxylic acids/esters/ethers, alkyl halides, and aliphatic/ aromatic amines. Eluted chemicals during GC-MS revealed the presence of nimbiol; 13-Methoxypodocarpa-8, 11,13-trien-3-one; Podocarpa-8, 11, 13-triene, 14-isopropyl-13-methoxy; 8-Chloro-5-quinolinecarboxylic acid; and 7-Chloro-2-cyclohexyl-4[3H]quinazolinone which has antimicrobial, antibacterial, anti-inflammatory, antifungal and analgesic properties validating the efficacy of the research formulation.