A simple HPLC procedure for the identification and quantitation of phytochelatins (PCs) in plant tissue extracts is described. The method, which does not require a derivatization step utilizes only 20 µL of sample volume. Linear quantitative response curve was generated for phytochelatin 3 (PC3) over a concentration range of 1.33 µmol/L-6.66 mmol/L. Linear regression analysis of the standard curve exhibited correlation coefficient of 0.996. Limit of detection (LOD) and limit of quantitation (LOQ) values were 0.1 and 0.5 µmol, respectively. Phytochelatin 3 recovery using this method was relatively high (above 85%). Intra-assay and inter-assay precision studies reflected a high level of reliability and reproducibility of the method. The applicability of the method for the quantitation of a wide range of PC concentrations in plant tissue extracts was demonstrated successfully.