高效液相串联质谱法监测女性风湿病患者备孕前血浆中特立氟胺浓度

目的:运用高效液相色谱串联质谱建立测定风湿病患者血浆中特立氟胺浓度的分析方法。方法:50μL血浆加入内标工作液200μL沉淀蛋白后稀释，再采用色谱柱Welch XB-C₁₈(2.1 mm×50 mm, 5μm)进行液相分离，流动相由10 mmol·L^-1甲酸铵溶液(A)和甲醇∶乙腈∶异丙醇=7∶1.5∶1.5(0.2%甲酸)(B)组成，梯度洗脱，流速0.7 mL·min^-1,分析时间2.5 min,进样量3μL,采用多反应监测，负离子模式，电喷雾离子化，监测离子对为m/z 268.4→82.0(特立氟胺)和m/z 272.9→82.0([2H4]-特立氟胺)。结果:特立氟胺在2～1 000 ng·mL^-1线性范围良好，各质控水平准确度为97.2%～105.5%,日内和日间相对标准偏差为4.4%～9.3%,回收率为93.4%～96.4%。结论:本方法专属性强、灵敏度和准确度高、耗时短、前处理简单，可应用于监测女性风湿病患者备孕前血浆中特立氟胺浓度。

Therapeutic monitoring of plasma teriflunomide concentration in female patients with rheumatic disease before pregnancy planning by LC-MS/MS

OBJECTIVE To establish a liquid chromatography-tandem mass spectrometry(LC-MS/MS)method for determining concentration of teriflunomide in plasma with rheumatic disease.METHODS 50 μL plasma sample was directly precipitated with internal standard,dissolved in acetonitrile.The sample was separated with Welch XB-C₁₈(2.1 mm×50 mm,5 μm),the mobile phase consisted of 10 mol·L^-1 ammonium formate solution(A)-methoanol∶acetonitrile∶ isopropanol(7∶1.5∶1.5,0.1% formic acid)(B)with gradient elution at the flow rate of 0.7 mL·min^-1,and the injection volume was 3 μL and analytical time was 2.5 min.Electrospray ion source,negative ion scanning,and multiple reaction monitoring mode was applied with ion pairs of m/z 268.4→82.0(teriflunomide)and m/z 272.9→82.0([2H4]-teriflunomide).RESULTS The linear range of teriflunomide was well accepted within 2-1000 ng·mL^-1,the relative standard deviation of intra-or inter-day precision was 4.4%-9.3% with a accuracy of 97.2%-105.5% among each quality control level,and the recovery was 93.4%-96.4%.CONCLUSION This method is specific,sensitive,accurate,short in time and simple in pretreatment.It can be used to monitor the concentration of terifluoramine in the plasma of female patients with rheumatism before pregnancy preparation.