Determination of Isoflavones in Radix puerariae from Different Origins by Ultra-High Performance Liquid Chromatography Based on Optimal Pretreatment Method

Abstract

A new method for simultaneous determination of puerarin, daidzin, daidzein and genistein in Radix puerariae by ultra-high performance liquid chromatography was established. The target analytes were extracted from Radix puerariae by 70% ethylene glycol with the assistance of ultrasonication, purified by the absorption of N-propyl ethylenediamine (PSA), and separated on a Supersil ODS column (4.6 mm × 250 mm × 2.5 μm). Gradient elution in 12 min was performed with the mobile phase 0.1% formic acid(A)–acetonitrile(B). The column temperature was 25 °C and the flow rate was 1 mL/min. The detection wavelength of the four target analytes was 250 nm. The limits of detection (LODs) of puerarin, daidzin, daidzein and genistein were 0.086 mg/L, 0.020 mg/L, 0.027 mg/L and 0.037 mg/L, respectively, and limits of quantitation (LOQs) were 0.29 mg/L, 0.065 mg/L, 0.090 mg/L and 0.12 mg/L, respectively. The recovery of the four substances ranged from 90.5% to 109.6%, and the relative standard deviation (n = 6) was less than 7.7%. With the established methods, puerarin, daidzin, daidzein and genistein in Radix puerariae from 11 origins were determined. The contents of the four compounds varied with the origin and variety. It provides basic data and technical means for quality control and regulation of Radix puerariae.