LC-MS/MS法测定人血清中地芬诺酯的浓度及其临床应用

LC-MS/MS determination of diphenoxylate in human blood and application in clinical study

目的:利用高效液相色谱-质谱联用仪(LC-MS/MS)建立血清中地芬诺酯的快速定性定量分析方法,为临床中毒患者的快速诊治提供依据。方法:选用Shim-pack XR-ODS色谱柱,以乙腈-10 mmol/L醋酸铵为流动相,采用梯度洗脱进行分离,流速:0.3 ml/min;柱温:35℃;进样量:5μl。样品用乙腈进行蛋白沉淀后进样,选用3200型质谱仪的多重反应监测(MRM)扫描方式进行检测。结果:地芬诺酯的线性范围为0.1 ng/ml～1000 ng/ml,定量下限和最低检测限分别为0.1 ng/ml和0.01 ng/ml。日内精密度RSD为2.7%,日间精密度RSD为3.8%。结论:本研究所建立的方法快速、灵敏,重复性好,适用于地芬诺酯中毒患者血液标本的快速定性定量检测。

Objective：To develop a LC-MS/MS method for the determination of the diphenoxylate in human serum and provide proof for the poisoners′ diagnosis.Methods：Diphenoxylate was extracted from serum by acetonitrile which was used as deproteinated solvent,and separated on a Shim-pack XR-ODS column maintained at 35℃ with acetonitrile and 10 mmol/L ammonium acetate as mobile phase by gradient elution.The flow rate was 0.30 ml/min and 5 μl aliquot of residues were injected into the LC-MS/MS system.Detection was carried out by multiple reaction monitoring on a 3200 LC-MS/MS system.Results：The assay was linear over the range 0.1 ng/ml～1000 ng/ml with a limit of quantitation of 0.1 ng/ml and a limit of detection of 0.01 ng/ml.The relative deviation of intra-and inter-day were 2.7%,3.8%.Conclusion：The method is a rapid,sensitive and reliable for the determination of diphenoxylate in human serum blood.The assay is applied to a fast qualitative and quantitative determination of diphenoxylate poisoners.