生物样品中苯乙醛酸和苯乙醇酸的HPLC测定方法

目的: 测定尿液中苯乙醛酸和苯乙醇酸含量。 方法: 采用Kromasil C₁₈ 柱(250 mm×4.6 mm,5 μm);流动相为乙腈-混合酸(冰醋酸0.5 mL+磷酸0.5 mL+水1000 mL)(5∶ 95),流速为1 mL·min^-1,柱温为25 ℃,检测波长为225 nm。 结果: 本实验对190个样品进行测定,PGA含量为0~0.384 mg·mL^-1,平均含量为0.071 mg·mL^-1;MA含量为0.007~0.497 mg·mL^-1,平均含量为0.121 mg·mL^-1。本方法苯乙醛酸在0.03298~0.4122 mg·mL^-1 (r=0.9991)范围内峰面积与浓度线性关系良好,定量限为17.2 ng;样品的加标回收率为95.2%~104.3%;批内精密度RSD为4.3%~6.8%,批间精密度RSD为4.5%~8.7%。苯乙醇酸在0.08158~1.0198 mg·mL^-1 (r=0.9994)范围内峰面积与浓度线性关系良好,定量限为20.4 ng;样品的加标回收率为99.3%~104.2%,批内精密度RSD为3.0%~5.1%,批间精密度RSD为5.7%~9.8%。 结论: 本方法操作简便,适用于尿中苯乙醛酸和苯乙醇酸的含量测定。

Objectives: To determine the concentrations of phenylglyoxylic acid and mandelic acid in urine by HPLC. Methods: A Kromasil C₁₈(250 mm×4.6 mm,5 μm) column was used.The mobile phase was acetonitrile-mixed acid (0.5 mL glacial acetic asid+0.5 mL phosphoric acid +1000 mL water) (5:95) at a flow rate of 1.0 mL·min^-1.The column temperature was 25 ℃,and the detection wavelength was set at 225 nm. Results: 190 samples were measured in this experiment,and results are as follows:PGA content of 0-0.384 mg·mL^-1,with an average content of 0.071 mg·mL^-1; the MA 0.007-0.497 mg·mL^-1,and the average content of 0.121 mg·mL^-1.A good linearity of PGA was obtained in the range of 0.03298-0.4122 mg·mL^-1 (r=0.9991)and the limit of quantification was 17.2 ng.The recoveries of spiked samples were in the range of 95.2%-104.3%,the within-run precision was between 4.3% and 6.8%,and the between-run precision was between 4.5% and 8.7%.A good linearity of MA was obtained in the range of 0.08158-1.0198 mg·mL^-1 (r=0.9994)and the limit of quantification was 20.4 ng.The recoveries of spiked samples were in the range of 99.3%~104.2%,the within-run precision was between 3.0% and 5.1%,and the between-run precision was between 5.7% and 9.8%. Conclusion: The method is simple and can be used for determination of phenylglyoxylic acid and mandelic acid in urine samples with satisfactory results.