RP-HPLC法测定人血清中天门冬氨酸、谷氨酸、甘氨酸和γ-氨基丁酸含量

Simultaneous determination of glutamate,aspartate,γ-aminobutyric acid and glycine in human serum by RP-HPLC

[目的]建立RP-HPLC法测定人体血清中天门冬氨酸、谷氨酸、甘氨酸和γ-氨基丁酸含量的方法。[方法]用邻苯二甲醛（OPA）柱前衍生;氨基酸的衍生物用Nova-Pak C18（3.9 mm×150 mm,4μm）分离;流动相A为30%甲醇、2%四氢呋喃、68%磷酸缓冲液（0.1 mol/L,pH 6.8）,流动相B为100%甲醇,用二元线性梯度程序洗脱;荧光检测器检测,激发光波长280 nm,发射光波长340 nm。[结果]在（4-400）nmol/mL范围内,各组分线性关系良好,各氨基酸的平均回收率为90.0%-93.1%。[结论]本方法操作简便、准确,适用于人体血清中天门冬氨酸、谷氨酸、甘氨酸和γ-氨基丁酸浓度的测定。

[ Objective] To develop a high performance liquid chromatography （HPLC） for simultaneous separation and rapid determination of four amino acid [ glutamate （ Glu）, aspartate （ Asp）, γ- aminobutyric acid （ GABA）, glycine （Gly） ] in human serum. [ Methods] The precolumn derivazation reagent was OPAo Derivatives were separated on a Nova - Pak C18 （3.9 mm×150 mm,4 μm）. Phosphate buffer solution, tetrahydrofuran and methanol as mobile phases, and gradient elution lasted 20 rain with fluorescence detection. [ Results] The range of calibration curve was （4 -400） nmol/mL. The calibrations were characterized by a coefficient of correlation higher than 0.997. The average extracted recovery was 90.0% - 93.1% , RSD% 〈 9.87%. [ Conclusion ] Our results showed that the method was rapid, accurate and simple.