UPLC-MS/MS法测定人血清中的神经内分泌肿瘤标志物5-羟基吲哚乙酸

Determination of 5-hydroxyindoleacetic acid in serum as neuroendocrine tumor marker by UPLC-MS / MS

目的采用超高效液相色谱串联质谱法测定人血清中5-羟基吲哚乙酸（5-HIAA）的含量。方法采用Waters Acquity UPLC BEH C18色谱柱（50 mm×2.1 mm,1.7μm）,流动相为2.5 mmol·L-1乙酸铵水溶液（含0.02%甲酸）-甲醇（68∶32）,流速0.2 m L·min-1,进样量3μL,柱温35℃,样品室温度6℃;采用电喷雾正离子模式电离,5-HIAA和甲氨蝶呤内标的监测离子分别为m/z 192.2→146.0、m/z 455.3→308.2。结果 5-HIAA的线性范围为0.0100~10.02μmol·L-1,平均回收率为92.1%~96.5%,批内、批间RSD均小于15%。结论所用方法的线性范围宽、分析时间短,可用于神经内分泌肿瘤的诊断及进展监测。

OBJECTIVE To establish an Ultrahigh performance liquid chromatographytandem mass spectrometry（ UPLC -MS/ MS） method for the determination of 5 - hydroxyindole acetic acid （ 5 - HIAA） in human serum. METHODS The concentrations of 5 -HIAA were analyzed on Waters Acquity UPLC BEH C18 column（50 mmx 2. 1 ram, 1.7 p,m）with mobile phase consisting of 2.5 mmol·L^-1 ammonium acetate（containing 0.02% aqueous formic acid） - methanol（68 ： 32） at a flow rate of 0.2 mL· min^-1. The column temperature was 35 ℃. The sample room temperature was set at 6 ℃ and the injection volume was 3 μL. The eleetorspray source and positive ion scan were used. 5 - HIAA and the international standard were monitored at （m/z） 192.2→146.0 and （m/z） 455.3→308.2,respectively. RESULTS The calibration curve of 5 - HIAA was linear over the range of 0.0100 - 10.02 μ mol· L^-1 The mean recoveries were from 92.1% to 96.5% . The intra - day and inter - day RSD were both less than 15%. CONCLUSION The method is sensitive with a wide assay range and short analysis time. It is suitable for the diagnosis and follow - up of neuroendocrine tumors （NETs） patients.