DEVELOPMENT AND VALIDATION OF HPLC METHOD FOR QUANTITATIVE ESTIMATION OF NUCIFERINE FROM LOTUS LEAVES

Nelumbo nucifera Gaertn (Family: Nelumbonaceae), commonly known as lotus, is an aquatic perennial plant. The lotus leaves are used to treat fever, hematemesis, hyperlipidaemia and obesity. The leaves are rich in alkaloids such as nuciferine, roemerine, pronuciferine and N-nornuciferine. The present study describes simple, accurate, precise, robust RP-HPLC method to detect and quantitate nuciferine as a marker from lotus leaves. The marker was resolved using Hemochrom Intsil C-18 column, with mobile phase methanol:10mm phosphate buffer (85:15), pH adjusted to 4 (using orthophosphoric acid) at a flow rate of 1 mL/min. Retention time of nuciferine was 5.60±0.2 minutes at 272 nm. Linear responses were obtained in concentration range of 1 - 9 ppm for nuciferine with correlation coefficient (r2) of 0.9996. The developed method was applied for quantitation of marker from lotus leaves. This method can be used to evaluate nuciferine containing extracts, thus conforming to the need of ensuring quality and safety of herbal drugs.