Design expert-supported development and validation of HPTLC method: An application in simultaneous estimation of quercetin and rutin in Punica granatum, Tamarindus indica and Prunus domestica
Aim: A simple, precise and accurate high performance thin layer chromatographic method has been developed for the simultaneous estimation of quercetin and rutin in Punica granatum, Tamarindus indica and Prunus domestica. Methods: The separation was carried out on Merck TLC aluminium sheets of silica gel 60 F254, (20 10 cm) with 250 mm thickness using toluene:ethyl acetate:formic acid (5:4:0.5, v/v/v) as a mobile phase. Results: The detection of quercetin and rutin was carried out at 297 nm. The drugs were satisfactorily resolved with Rf values of 0.44 0.02 and 0.59 0.02 for quercetin and rutin, respectively. The accuracy and reliability of the method was assessed by evaluation of linearity (100e1000 ng/spot for quercetin and 500e3000 ng/spot for rutin). Intra- and inter-day precision measured as coefficient of variation were less than 2% for both analytes. The limits of detection and quantification were found to be 33.54 and 101.66 ng/spot, respectively for quercetin and 367.21 and 1112.75 ng/spot, respectively for rutin. Conclusion: The proposed method was found to be accurate, precise, reproducible and specific and can be applicable for the simultaneous determination of quercetin and rutin in samples. Key words: HPTLC, Punica granatum, Tamarindus indica, Prunus domestica, Simultaneous.