Quantify biomarkers in Asthimajja Pachak Kwath by High Performance Thin Layer Chromatography

Abstract High Performance Thin Layer Chromatography has capacity to run many samples simultaneously and its reduced solvent usage. Simultaneous quantification of gallic acid, ferulic acid, rutin and berberine in Asthimajja Pachak Kwath has been developed and validated using a specific, precise, accurate, and low-cost high performance thin layer chromatography technique. Toluene: ethyl acetate: chloroform: methanol: formic acid (5:2:2:2:1, v/v/v/v/v) was used as mobile phase during separation, which took place on a stationary phase of pre-coated silica gel aluminum plate 60 F 254 . At 270 nm, separated components were densitometrically measured. The method was validated for linearity and range, precision, reproducibility, specificity, robustness, accuracy, limit of detection and limit of quantification as per International Council for Harmonization Q2 (R2) guideline. Gallic acid, ferulic acid, rutin and berberine regression coefficients ( R 2 ) were found to be 0.9995, 0.9990, 0.9985 and 0.9997 accordingly. Gallic acid, ferulic acid, rutin and berberine percentage recoveries were measured to be 99.81 ± 0.833, 99.15 ± 0.776, 99.56 ± 0.582 and 99.90 ± 0.719 respectively. Having a reliable analytical method is essential for ensuring the quality, consistency, and efficacy of herbal formulations in traditional medicine. This meticulous approach is crucial for accurate quantification of the target compounds in the Asthimajja Pachak Kwath .