Abstract: OBJECTIVE To establish a reversed-phase high-performance liquid chromatographic method derivated by PITC (phenylisothiocyanate) pre-column for the determination of 17 amino acids content of fresh, heart, lung and intestines in Elaphe carinat (Cauliflower), and to develop the edible and medical value of Elaphe carinat further. METHODS Using Waters PICO·TAG^TM column(3.9 mm×150 mm, 5 μm), gradient elution with sodium acetate buffer as mobile phase A, acetonitrile-water (60∶40) as mobile phase B, detection wavelength of 254 nm. RESULTS All of 17 amino acids were well separated within 20 min; 17 amino acids had a good linear relationship in their respective concentration ranges (0.997 6<r<0.999 9). The average recoveries were in 98.3%-107.5%. CONCLUTION This method is simple, accurate and reliable with a good reproducibility and can be used to determine amino acids content of snake fresh, heart, lung and intestines in Elaphe carinat, which provides experimental evidence of the further development, usage and protection of the Elaphe carinat resource.