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建立以PITC法和AQC法为柱前衍生试剂测定血液和尿液中游离氨基酸含量的测定方法。采用Waters-e2695操作系统,色谱柱为Shim-vp,ODS(250mm×4.6mm,5μm)(日本,岛津公司),以甲醇/乙腈/水和醋酸钠溶液(pH 6.5)为流动相,梯度洗脱。分别采用紫外和荧光检测器对血液和尿液中游离氨基酸进行含量测定。结果显示,两种衍生化方法灵敏度好、分离度高,具有良好的线性范围(r>0.990 0),准确度高(平均回收率为75.1%～127.0%),进样精密度好(RSD为0.12%～3.42%)。PITC法在尿液中游离氨基酸含量测定中显示了良好的测试准确性;而AQC法测定尿液中组氨酸、苏氨酸、脯氨酸超出线性范围,需要对尿样的前处理进行深入研究。
A method for determination of free amino acids in blood and urine by PITC method and AQC method was established. The Waters-e2695 operating system was used. The mobile phase was Shim-vp, ODS (250 mm × 4.6 mm, 5 μm) Elution. The free amino acids in blood and urine were determined by ultraviolet and fluorescence detector respectively. The results showed that the two derivatization methods had good sensitivity, high resolution, good linearity (r> 0.990 0), high accuracy (average recovery 75.1% ~ 127.0%), good precision of injection 0.12% ~ 3.42%). PITC method in the determination of free amino acids in urine showed good test accuracy; and AQC method for determination of urine Histidine, threonine, proline beyond the linear range, the need for pre-treatment of urine samples the study.