Objective To investigate the change of plasma L-threonine (L-Thr) ,D-threonine (D-Thr) ,L-alanine (L-Ala) and D-alanine (D-Ala) in the patients with primary hepatocellular carcinoma (HCC) by using the reversed-phase high-performance liquid chromatography (RP-HPLC) .Methods The o-phthaldialdehyde (OPA) and N-acetyl-L-cysteine (NAC) column with pre-column derivatization was adopted to perform the separation .The mobile phase was 20 mmol/L sodium dihydrogen phosphate solution and acetonitrile with a programmed gradient elution and the flow rate of 1 .0 mL/min .The excitation wavelength was set at 350 nm and the emission wavelength was set at 450 nm .Results The threonine and alanine enantiomers could be simultaneously determined by this method .The regression equation showed the good linear correlation The limits of detection (LOD) for L-Thr ,D-Thr ,L-Ala and D-Ala were 11 .4 pg/mL ,9 .77 pg/mL ,4 .11 pg/mL and 3 .50 pg/mL respectively .The recovery rate was 90 .22% -105 .7% .With-in-day and between-day RSD was 1 .04% -4 .11% .Conclusion The threonine and alanine enantiomers could be simultaneously de-termined by this method .The regression equation showed the good linear correlation The limits of detection (LOD) for L-Thr ,D-Thr ,L-Ala and D-Ala were 11 .4 pg/mL ,9 .77 pg/mL ,4 .11 pg/mL and 3 .50 pg/mL respectively .The recovery rate was 90 .22%-105 .7% .Within-day and between-day RSD was 1 .04% -4 .11% .%目的：用反相高效液相色谱法探讨原发性肝癌患者L-苏氨酸（L-threonine ，L-Thr）、D-苏氨酸（D-threonine ，D-Thr）、L-丙氨酸（L-alanine ，L-Ala）和D-丙氨酸（D-alanine ，D-Ala）水平的变化。方法采用邻苯二甲醛和N-乙酰-L-半胱氨酸柱前衍生用反相色谱柱进行分离，流动相为20 mmol/L磷酸二氢钠溶液、乙腈梯度洗脱，流速1．0 mL/min ，激发波长350 nm ，发射波长450 nm。结果该方法可以同时分离测定血液中苏氨酸、丙氨酸氨基酸对映体，回归方程线性相关性良好，检测线分别为11．4 pg/mL、9．77 pg/mL、4．11 pg/mL和3．50 pg/mL ，回收率为90．22％～105．7％，日间、日内相对标准偏差（RSD）为1．04％～4．11％。结论该方法准确、灵敏、适用于临床和科研。
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