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 evaporative light-scattering detector(ELSD). GSH, Phytochelatin 2 (PC2), phytochelatin 3 (PC3), phytochelatin 4 (PC4), phytochelatin 5 (PC5) and phytochelatin 6 (PC6) can be separated with baseline separation within 9 minutes using a Venusil AA column (250 mm × 4.6 mm i.d., 5 μm particle sizes). The acetonitrile (A) and water containing 0.1% trifluoroacetic acid (0.1% TFA, B) were employed as the mobile phase for the gradient elution. The drifttube temperature and flow rateofcarriergas (N2) were 50℃ and 1.5 L·min-1, respectively. Under the 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%.