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

摘要

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%.