尿中苯乙醛酸和苯乙醇酸测定的超高效液相色谱串联质谱法

Determination of phenylglyoxylic acid and mandelic acid in urine by ultra high performance liquid chromatography tandem mass spectrometry

目的建立尿中苯乙醛酸(PGA)和苯乙醇酸(MA)的超高效液相色谱串联质谱分析方法。方法于2022年4月开展研究,尿样经初始流动相直接稀释,过膜后以Waters HSS T3色谱柱分离,在负电离模式(ESI-)、多反应监测(MRM)条件下分析,外标法定量测定人尿中PGA、MA的含量。结果PGA和MA的测定在10~1000 ng/ml的范围内线性关系良好,相关系数r为0.9999。PGA的线性回归方程为y=1141.4x+2157.3,方法检出限和定量下限为0.081 ng/ml和0.269 ng/ml,加标回收率为90.47%~99.83%;MA的线性回归方程为y=62.8x+140.3,方法检出限和定量下限为0.551 ng/ml和1.836 ng/ml,加标回收率为92.75%~101.09%;PGA和MA的批内精密度和批间精密度均<5%。结论建立了尿中PGA和MA测定的超高效液相色谱串联质谱分析法,样品前处理操作简单,方法准确度和精密度均满足标准要求,可用于普通人群和职业接触人群尿中PGA和MA的监测评估。

Objective To develop a method for the analysis of phenylglyoxylic acid(PGA)and mandelic acid(MA)in urine by ultra-high performance liquid chromatography tandem mass spectrometry.Methods The study was conducted in April 2022.Urine samples were directly diluted with the initial mobile phase,separated by Waters HSS T3 column after passing through the membrane,and analyzed under negative ionization mode(ESI-)and multiple reaction monitoring(MRM)conditions,the contents of PGA and MA in human urine were quantitatively determined by external standard method.Results The determination of PGA and MA showed a good linear relationship within the range of 10-1000 ng/ml,with a correlation coefficient of 0.9999.The linear regression equation of PGA was y=1141.4x+2157.3,the detection limit and lower limit of quantification of the method were 0.081 ng/ml and 0.269 ng/ml,and the recovery rate was 90.47%-99.83%.The linear regression equation of MA was y=62.8x+140.3,the detection limit and lower limit of quantification of the method were 0.551 ng/ml and 1.836 ng/ml,and the recovery rate was 92.75%-101.09%.The intra and inter batch precision of PGA and MA were both<5%.Conclusion An ultra-high performance liquid chromatography tandem mass spectrometry method for the analysis of PGA and MA in urine was established.The sample pretreatment operation is simple,and the accuracy and precision of the method meet the standard requirements.It can be used for monitoring and evaluating PGA and MA in urine of the general population and occupational contact population.