Simultaneous determination of apoglaziovin, caaverin, n-metylasimilobin in the leaves of Nelumbo nucifera (Gaertn.) (Nelumbonaceae) by HPLC-PDA

Abstract

 A reversed-phase HPLC method for simultaneous determination of apoglaziovin, caaverine, and N-methyl asimilobine in the leaves of Nelumbo nucifera (Gaertn.) (Nelumbonaceae) was successfully developed as follows: HPLC System - Phenomenex Gemini  NX; Column - RP-C18 (30 x 3.0 mm; 3 μm); Detector – UV (272 nm); Mobile phase - acetonitrile-0.1 % triethylamine solution adjusted to pH 8.5 by 5 % acid acetic solution, in gradient mode; Flow rate - 0.8 mL/min, By validation observing the ICH guideline, the method showed the linearity range of 5 – 200 ppm (R² > 0.999), high selectivity, high accuracy (94.6 - 104.7 %, RSD 1.05 - 1.97) and precision (RSD < 5 %), high sensitivity (LOD: 0.03 – 0.3 ppm, LOQ: 0.1 - 1.15 ppm). For practical tests, alcaloids in the leaves of Nelumbo nucifera collected from 12 Vietnamese provinces were simultaneously determined. Significant quantitative differences were revealed one from another growing area and season as well. Especially, the samples from Dong Thap province predominantly contained apoglaziovin and N-methyl asimilobine, whereas those from the central provinces showed a higher relative occurrence of caaverine.