Separation and Quantification of Tannic Acid in Bryophyllum pinnatum (Lam.) Kurz. by High Performance Thin Layer Chromatography

Abstract

A sensitive, selective and robust quantitative densitometric high-performance thin layer chromatographic method was developed and validated for the determination of tannic acid in the different parts of Bryophyllum pinnatum (Lam.) Kurz. Tannic acid was used as a chemical marker for the standardization of Bryophyllum pinnatum (Lam.) Kurz. plant extracts. The separation was performed on silica gel 60 F₂₅₄ TLC plates using toluene-ethyl acetate-formic acid (5:4:1 v/v/v) as mobile phase. The quantitation of tannic acid was carried out using the densitometric reflection/absorption mode at 270 nm. A precise and accurate quantification can be performed in the linear working concentration range of 20-100 μg mL^-1 with good correlation (r² = 0.997). The method was validated for, precision, recovery, robustness, limit of detection (LOD) and quantitation (LOQ) as per ICH guidelines. Specificity of quantitation was confirmed using retention factor (R_f), UV spectral correlation of marker compound (tannic acid) in sample track.