HPLC-DAD separation and determination of major active constituents in an important Tibetan medicine Meconopsis quintuplinervia from different regions of Qinghai-Tibet Plateau

Abstract

This study describes a simple and sensitive method for the simultaneous determination of major flavonoids active constituents in a traditional Tibetan medicine Meconopsis quintuplinervia collecting from 29 different localities of Qinghai-Tibet Plateau by high-performance liquid chromatography coupled with diode array detection (HPLC-DAD). The optimal conditions of separation and detection were achieved on a Zorbax Eclipse XDB-C18 column (4.6 × 250 mm, 5 μm) with a gradient of methanol and 0.05% aqueous acetic acid (v/v), at a flow rate of 1.0 mL min-1, detected at 370 nm. The complete separation was obtained within 40 min for the four analytes. The recovery of the method was >100%, and all the flavonoids showed good linearity (r≥0.9990) in a relatively wide concentration range. The limits of detection (LOD) are (injection volume 10 μL, at a signal-to-noise ratio of 3, S/N = 3:1) between 0.09 and 0.36 μg mL-1. Furthermore, the main characteristics of flavonoids in M. quintuplinervia were also analyzed by principle component analysis (PCA).   Key words: HPLC-DAD, flavonoids, Meconopsis quintuplinervia, Tibetan medicine.