A three column-switching high-performance liquid chromatography (HPLC) using an electrochemical detector (ECD) equipped with a diamond electrode was established to determine 3-hydroxypropylmercapturic acid (3-HPMA) in urine. An extracted urine sample was consecutively fractionated using a strong anion-exchange column (first column) and a C8 column (second column) via a switching valve before application on an Octa Decyl Silyl (ODS) column (third column), followed by ECD analysis. The% recovery of 3-HPMA standard throughout the three-column process and limit of detection (LOD) were 94 ± 1% and 0.1 pmol, respectively. A solid phase extraction step is required for the sensitive analysis of 3-HPMA in urine by column-switching HPLC-ECD despite a decreased% recovery (55%) of urine sample spiked with 100 pmol of 3-HPMA. To test the utility of our column-switching HPLC-ECD method, 3-HPMA levels of 27 urine samples were determined, and the correlation between HPLC-ECD and LC-Electrospray ionization (ESI)-MS/MS method was examined. As a result, the median values of μmol 3-HPMA/g Creatinine (Cre) in urine obtained by column-switching HPLC-ECD and LC–MS/MS were 2.19 ± 2.12 μmol/g Cre and 2.13 ± 3.38 μmol/g Cre, respectively, and the calibration curve (y = 1.5171x − 1.007) exhibited good linearity within a defined range (r² = 0.907). These results indicate that the combination of column-switching HPLC and ECD is a powerful tool for the specific, reliable detection of 3-HPMA in urine.

建立了使用配备了金刚石电极的电化学检测器（ECD）的三柱切换高效液相色谱（HPLC），用于测定尿液中的3-羟丙基硫醇酸（3-HPMA）。使用强阴离子交换柱（第一根柱）和C8柱（第二根柱）通过切换阀将分出的尿液样品连续分级，然后再施加到Octa Decyl Silyl（ODS）柱（第三根柱）上，然后进行ECD分析。整个三柱过程中3-HPMA标准品的回收率％和检出限（LOD）分别为94±1％和0.1 pmol。尽管通过100 pmol 3-HPMA加标的尿液样品的回收率降低了（55％），但通过柱切换HPLC-ECD进行尿液中3-HPMA的灵敏分析仍需要固相萃取步骤。为了测试我们的柱切换HPLC-ECD方法的实用性，测定了27个尿液样品的3-HPMA水平，并检查了HPLC-ECD和LC-电喷雾电离（ESI）-MS / MS方法之间的相关性。结果，通过柱切换HPLC-ECD和LC-MS / MS获得的尿液中μmol3-HPMA / g肌酐（Cre）的中位数为2.19±2.12μmol/ g Cre和2.13±3.38μmol/ g Cre ，并且校正曲线（y = 1.5171x − 1.007）在定义的范围内显示出良好的线性度（r ²  = 0.907）。这些结果表明，色谱柱转换HPLC和ECD的结合是用于尿液中3-HPMA特异性，可靠检测的强大工具。