Development and Validation of HPTLC Method for Quantitative Estimation of Nuciferine from Lotus leaves
Department of Quality Assurance, BharatiVidyapeeth’s College of Pharmacy, C.B.D. Belapur, Navi Mumbai-400 614, India
Department of Quality Assurance, BharatiVidyapeeth’s College of Pharmacy, C.B.D. Belapur, Navi Mumbai-400 614, India, E-mail: [email protected]
Nelumbo nucifera (Nelumbonaceae), commonly known as lotus, is an aquatic perennial plant. A simple, precise and accurate high-performance thin-layer chromatography method for quantitative estimation of an alkaloid, nuciferine from methanol extract of lotus leaf was developed. The method employed high-performance thin-layer chromatography aluminium plate precoated with silica gel 60F254 (10x10 cm) as stationary phase and separation was achieved by using a suitable mobile phase chloroform:ethylacetate:methanol:glacial acetic acid [6.5:1:2.5:0.025 v/v/v/v] and dragendorff’s reagent was used for detection. Quantitation of the method was achieved by densitometric method at 272 nm using Camag TLC scanner 3. This method showed good separation of nuciferine in the extract with Rf value 0.44±0.02. The regression analysis data indicated good linear relationship for the calibration plots for nuciferine in the range of 100-450 ng/spot and regression coefficient was 0.998. The accuracy and reliability of the proposed method was ascertained by evaluating various validation parameters like linearity, precision, accuracy, robustness, specificity, limit of quantification and limit of detection according to the ICH guideline. Statistical analysis of the data showed that the method is reproducible and selective for the estimation of nuciferine. The developed high-performance thin-layer chromatography method can be used for quantification of nuciferine in different species of lotus and formulations containing nuciferine.