生物样品中苯乙醛酸和苯乙醇酸的HPLC测定方法

HPLC determination of phenylglyoxylic acid and mandelic acid in biological samples

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,5μm);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 Cls （250 mm ×4.6mm,5μm） 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 sam- ples were measured in this experiment, and results are as follows ：PGA content of 0 -0. 384 mg·mL^-1,with an av- erage 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. 0198mg·mL^-1 （ r = 0. 9994 ） and the lim- it of quantification was 20. 4 ng. The recoveries of spiked samples were in the range of 99.3% - 104.2% , the with- in -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.