Development of the HPLC-ELSD method for the determination of phytochelatins and glutathione in Perilla frutescens under cadmium stress conditions

Abstract

A rapid, accurate and simple method was developed for the simultaneous determination of glutathione (GSH) and phytochelatins (PCs) by high-performance liquid chromatography (HPLC) with an evaporative light-scattering detector. GSH, phytochelatin 2 (PC2), PC3, PC4, PC5 and PC6 can be separated with baseline separation within 9 min using a Venusil AA column (250 mm × 4.6 mm i.d., 5 µm particle sizes). Acetonitrile and water containing 0.1% trifluoroacetic acid (0.1%) were employed as the mobile phase for the gradient elution. The drift tube temperature and flow rate of the carrier gas (N2) were 50°C and 1.5 l min−1, respectively. Under optimum conditions, good linear regression equations of six analytes were obtained with the detection limits ranging from 0.2 to 0.5 µg ml−1. The proposed method has been applied successfully for the quantification of GSH and PCs in Perilla frutescens (a cadmium hyperaccumulator) under cadmium stress. The recoveries were between 82.9% and 115.3%.