Development and Validation of a High-Performance Thin-Layer Chromatography Method for Simultaneous Quantification of...

A validated high-performance thin-layer chromatography (HPTLC) method was developed for the simultaneous quantification of bergenin and oleanolic acid in Pashanbhed (Bergenia ligulata), Lambadi (Celosia argentea) and Kalipath (Cissampelos pareira) for quality assessment. The developed HPTLC method employed a mobile phase of toluene, ethyl acetate, methanol and formic acid (4:5:1:2 v/v) on silica gel 60F254 plates. The plates were scanned at 254 and 366 nm using a Camag Scanner IV with vision CATS software. The method was validated for linearity, accuracy, precision, specificity, limit of detection and limit of quantification. The linearity range for bergenin and oleanolic acid was 4-20 and 10-50 μg/spot, respectively, with correlation coefficients of 0.9998. The average recoveries for bergenin and oleanolic acid were 98.75-99.94% and 98.62-100.83%, respectively. The bergenin content in the methanolic extract and crude drug of B. ligulata was 4.8% and 1.25% w/w, respectively. The oleanolic acid content in the methanolic extract of C. argentea and C. pareira was 1.98% and 1.55%, while in the crude drug, it was 0.24% and 0.16% w/w, respectively. The developed HPTLC method is simple, accurate and reproducible, making it suitable for routine analysis, standardization and quality control of bergenin and oleanolic acid in various natural herbs and Ayurvedic formulations.