生物检材中34种常见毒品及代谢物的液相色谱轨道阱质谱筛查方法研究

The Screening Approaches for 34 Common Drugs and Metabolites in Biological Samples by Liquid Chromatography Orbital Trap Mass Spectrometry

目的建立高效液相色谱轨道阱质谱对生物检材中34种常见毒品及代谢物的筛查方法。方法尿样和血样经乙酸乙酯提取后,氮吹浓缩和复溶;毛发样品依次用水和丙酮振荡洗涤、晾干后剪成约1 mm,置冷冻研磨仪中粉碎,甲醇提取,过滤后,滤液供仪器分析。色谱分析采用Hypersil Gold PFP色谱柱(2.1 mm×100 mm,3μm)分离,甲醇-5 mmol/L乙酸铵溶液作为流动相,梯度洗脱,流速为400μL/min。质谱分析采用电喷雾正负离子交替模式,使用一级母离子全扫描和数据依赖的二级子离子扫描(Full MS/dd-MS^(2))模式,采用Xcalibur4.0软件分别对仪器和数据进行控制和采集,利用TraceFinder3.3软件进行筛查识别。结果血样、尿样和发样的34种常见毒品及其代谢物方法检出限分别为3.30~10700 ng/L、4.43~5440 ng/L和0.0350~4.21μg/kg。在5.0、10、20μg/L加标水平下,日内精密度为3.50%~6.00%,日间精密度为4.18%~9.90%。采集了尿样、血样和发样共1125份生物检材,采用该方法进行筛查分析,阳性样本中有87.9%为男性,96.7%的吸毒者吸食单一毒品,3.3%吸毒者吸食混合毒品,吸食毒品种类从高到低依次为甲基苯丙胺(75.8%)、海洛因(18.5%)、氯胺酮(2.4%)、其他毒品(3.3%)。与高效液相色谱三重四极杆质谱分析结果比较,该方法能同时识别出更多毒品、实现回顾性分析。结论建立的方法简便、灵敏度良好,适用于生物检材中常见毒品及代谢物的筛查分析。

Objective To establish a high-performance liquid chromatography orbital trap mass spectrometry(HPLC-Obitrap MS)method for screening 34 common drugs and metabolites in biological samples.Methods The target analytes in urine and blood samples were extracted with ethyl acetate,concentrated by nitrogen blowing and redissolved.The hair samples were washed with water and acetone,dried and cut into bits of about 1 mm,and then crushed in a freezing grinder.The analytes were extracted with methanol,and after filtration,the filtrate was used for instrumental analysis.Hypersil Gold PFP(2.1 mm×100 mm,3μm)column was used for chromatographic separation.Methanol and 5 mmol/L ammonium acetate solution were used as mobile phase with gradient elution at a flow rate of 400μL/min.Mass spectrometry was done by electrospray positive and negative ion alternation mode.The data were collected using Full MS and Full MS/dd-MS^(2)mode.Xcalibur 4.0 software was used to control instruments and to collect data,and TraceFinder 3.3 was used for screening and identification.Results The method’s detection limits for 34 drugs and their metabolites in blood,urine and hair samples were 3.30-10700 ng/L,4.43-5440 ng/L,0.0350-4.21μg/kg,respectively.The intra-day and inter-day precisions of the spiked samples at the levels of 5.0,10,and 20μg/L were 3.50%-6.00%and 4.18%-9.90%,respectively.A total of 1125 biological samples of urine,blood and hair were collected and screened.The results showed that 96.7%of the drug users were taking a single drug,while 3.3%were mixed drug users.The main types of drug of abuse were methamphetamine(75.8%),heroin(18.5%),ketamine(2.4%)and other drugs(3.3%),and 87.9%of the positive samples were from male users.Compared with the results of high-performance liquid chromatography triple quadrupole mass spectrometry,this method can be used to identify more types of drugs in one run and to conduct retrospective analysis.Conclusion The method established in the study is simple and sensitive and is well suited for the screening of common drugs and metabolites in biological samples.