Bauhinia variegata

Bauhinia variegata Linn. (Fabaceae) is a widely used medicinal plant whose bark exhibits antioxidant, anti-inflammatory, antimicrobial, hepatoprotective, and anticancer activities, largely attributed to its flavonoid content, of which quercetin is a key bioactive marker. Since no validated method previously existed for quantifying quercetin specifically in B. variegata bark, the present study aimed to develop and validate a simple, sensitive, and reproducible HPTLC method for this purpose, in accordance with ICH Q2(R1) guidelines. Chromatographic separation was achieved on silica gel 60 F₂₅₄ plates using a mobile phase of ethyl acetate: toluene: methanol: formic acid (3.5: 6: 0.5: 0.5, v/v/v/v), with densitometric detection at 383 nm, giving a well-resolved band at Rf 0.54. The method showed good linearity over 100–700 ng/band (y = 23.063x − 430.75, R² = 0.991), with an LOD of 20 ng/band and LOQ of 80 ng/band. Precision (%RSD < 2% for repeatability, intraday, and interday studies), accuracy (mean recovery 98.57–100.65%), robustness, and specificity all met acceptance criteria, confirming the reliability of the method. Application to the methanolic bark extract of B. variegata revealed a quercetin content of 0.012% w/w, with consistent results across replicate analyses. This validated HPTLC method offers a rapid, cost-effective, and reliable tool for the routine quality control, standardization, and phytochemical evaluation of B. variegata bark and its extracts. Keywords: Quercetin; HPTLC; Bauhinia variegata; method validation; ICH Q2(R1); phytochemical standardization

Bauhinia variegata (Mountain ebony) is a well known plant in traditional medicine which belongs to the family Cesalpiniaceae. It is widely used for various ailments like skin diseases, tumour, ulcers and various infections. Against this backdrop the present study attempts to scientifically evaluate the analgesic, antifungal, anti-tumour potential of this plant. The ethanolic extract of the plant was prepared by soxhlation. Acute toxicity studies carried out on the extract revealed that it was safe at the dose of 2000 mg/kg body weight. The extract was then evaluated for its analgesic activity by eddy’s hot plate method. The antifungal evaluation was carried against Candida albicans and Aspergillus fumigatus by disc diffusion method in Sabouraud agar medium. The extract was studied for short term in-vitro cytotoxicity against DLA cell lines by Trypan blue assay method. The ethanolic extract of Bauhinia variegata bark exhibited significant analgesic activity at both the doses of 200 and 400 mg/kg body weight (p