Simultaneous determination of lupeol and β-sitosterol by high-performance thin-layer chromatographic method inC rataeva nurvala Buch-Ham. stem bark

Crataeva nurvala Buch-Ham. (Capparidaceae) is an important medicinal plant, the bark of which is extensively used in Ayurveda as a remedy for kidney stones, urinary tract infections, prostate-related disorders, and benign prostatic hypertrophy. In this study, a sensitive and reliable high-performance thin-layer chromatography (HPTLC) method has been developed for the simultaneous quantification of two bioactive markers, lupeol and beta-sitosterol, in the stem bark ofC. nurvalausing precoated silica gel G 60 F-254 HPTLC plates as the stationary phase and toluene-methanol (9.6:0.4,v/v) as the mobile phase. Detection and quantification were carried out by densitometric scanning at 545 nm after derivatization using 1% anisaldehyde-sulfuric acid reagent. The method was validated for linearity, inter-day and intra-day precision, repeatability, accuracy, specificity, limit of detection, and limit of quantification. The linear regression analysis data for the calibration plots showed a good linear relationship (r(2) = 0.987-0.990) in the concentration range of 2 to 6 mu g for lupeol and beta-sitosterol, both with respect to areas. The proposed method provides good resolution of lupeol and beta-sitosterol from other constituents present in the stem bark of C. nurvala. It was found to be simple, accurate, precise, and reproducible and would potentially be a useful tool for routine analysis of C. nurvala.