UHPLC-HRMS快速测定血液中43种化学药物

Rapid Determination of 43 Chemical Drugs in Blood with UHPLC-HRMS

目的建立血液中43种化学药物快速测定的UPLC-HRMS方法,并应用于测定100份杭州地区非正常死亡人员的血液,分析43种化学药物在血液中的含量情况。方法以乙腈沉淀蛋白,离心后取上清液,氮气流下吹干,残渣以初始流动相复溶,离心后上清液用0.22μm微孔滤膜过滤,供仪器分析;采用C18(150 mm×2.1 mm,3.0μm)色谱柱分离,流动相:0.1%甲酸+5 mmol甲酸铵(A相),甲醇+0.1%甲酸(B相),梯度洗脱;质谱检测,采用电喷雾离子源(HESI-II),正负离子同时扫描,实时二级质谱扫描(Full MS/dd MS2)。结果43种化学药物在10~1000 ng·mL^(-1)内线性良好,r^(2)=0.9910~0.9998;定量下限为10 ng·mL^(-1);基质效应为79.4%~99.7%;方法回收率为80.4%~97.0%;日内精密度RSD为0.1%~3.2%;日间精密度RSD为0.6%~9.8%。以该法测定杭州市公安司法鉴定中心全血样品100例,检出2例,均为西布曲明。结论本方法准确、灵敏,适用于全血中43种化学药物快速检测。

OBJECTIVE To establish a UPLC-HRMS method for simultaneous determination of 43 chemical drugs in blood and apply the method to determine the content of 43 chemical drugs in blood of 100 abnormal death from Hangzhou.METHODS Proteins were precipitated using acetonitrile.After centrifugation,the supernatant was collected and dried under a nitrogen stream.The residue was reconstituted with the initial mobile phase,centrifuged,and the supernatant was filtered through a 0.22μm microporous membrane for instrument analysis.Separation was performed using a C_(18)(150 mm×2.1 mm,3.0μm)chromatographic column.The mobile phase consisted of 0.1%formic acid+5 mmol ammonium formate(phase A)and methanol+0.1%formic acid(phase B)with gradient elution.Mass spectrometric detection was carried out using an electrospray ionization source(HESI-II)with simultaneous positive and negative ion scanning and real-time tandem mass spectrometry scanning(Full MS/ddMS2).RESULTS The linearity of 43 chemical drugs within the range of 10 to 1000 ng·mL^(−1) was good,with r2 ranging from 0.9910 to 0.9998;the limit of quantification was 10 ng·mL^(−1);the matrix effect was between 79.4%and 99.7%;the method recovery rate was between 80.4%and 97.0%;the intra-day precision(RSD)ranged from 0.1%to 3.2%;and the inter-day precision(RSD)ranged from 0.6%to 9.8%.Using this method,100 whole blood samples from the Hangzhou Public Security Judicial Identification Center were tested,with 2 samples detected,both of which contained sibutramine.CONCLUSION The method is sensitive and accurate for fast detection of 43 chemical drugs in human blood.