Validated HPTLC method for simultaneous quantification of sennoside A, sennoside B, and kaempferol in Cassia fistula Linn

Abstract

This paper describes a simple, precise, and accurate HPTLC method for simultaneous quantification of sennoside A, sennoside B, and kaempferol in Cassia fistula whole plant extract. Chromatographic separation of the sample extract was performed on aluminium foil plates coated with silica gel 60 F 254 as stationary phase. The mobile phase was toluene-ethyl acetate-methanol-formic acid 8:10:5:2 ( υ/υ ). Densitometric evaluation of the separated bands was performed at 270 nm. Sennosides A and B and kaempferol were satisfactorily resolved at R F 0.22 ± 0.05, 0.19 ± 0.05, and 0.81 ± 0.05, respectively. Recovery of sennosides A and B and kaempferol from Cassia fistula extract was 98.03, 98.74, and 99.08%, respectively. The method was validated for specificity, accuracy, linearity (100-400 ng per band), and precision (instrument precision in the range 1.03-1.33 and method precision in the range of 1.31-1.75) in accordance with ICH guidelines.