UPLC-HRMS法快速测定血液中43种化学药物的研究

Development of UPLC-HRMS assay for simultaneous determination of 43 chemical drugs in human blood

关键词:  法医毒物分析  化学药品  超高效液相色谱-高分辨质谱  血液

Key words:  Forensic toxicological analysis  chemical drug  UPLC-HRMS  blood

摘要:

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微孔过滤膜过滤，供仪器分析；采用C18色谱柱（150mm×2.1mm，3.0μm）分离，流动相：0.1%甲酸+5mmol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/ddMS2）。结果 43种化学药物在10~1000ng/mL范围内线性良好，r2 =0.9910~0.9998；定量下限为10ng/mL；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、快捷、灵敏适用于全血中43种化学药物快速检测。

Abstract:

OBJECTIVE To establish a UPLC-HRMS assay for simultaneous determination of 43 chemical drugs in human blood and apply the method to determine the content of 43 chemical drugs in blood of 100 samples from Hangzhou. METHODS Acetonitrile was added to the sample for precipitating the protein, the mixture was centrifuged for 10min at 4℃, 14000r/min, the supernatant was filtered, and then the filtrate was analyzed with UPLC-HRMS. The separation was performed on C18 chromatography column (150mm×2.1mm，3.0μm), the mobile phase was consisted of 0.1% formic acid + 5mmol ammonium formate (phase A), Methanol + 0.1% formic acid(phase B) with gradient elution. RESULTS revealed the calibration curves for 43 chemical drugs were linear over the ranges of 10～1000ng/mL, r2 =0.9910~0.9998, the method recoveries were 80.4%~97.0% , the intraday and interday precious were 0.1%～3.2% and 0.6%～9.8% respectively. The method was applied to analyze 100 human samples stored by Center of Forensic Sciences of Hangzhou. CONCLUSION he method is sensitive and accurate for fast detection of 43 chemical drugs in human blood.