Karinen Ritva, Andersen Jannike Mørch, Ripel Ase, Hasvold Inger, Hopen Anita Braute, Mørland Jørg, Christophersen Asbjørg S
Division of Forensic Toxicology and Drug Abuse, Norwegian Institute of Public Health, P.O. Box 4404, Nydalen, 0403 Oslo, Norway.
J Anal Toxicol. 2009 Sep;33(7):345-50. doi: 10.1093/jat/33.7.345.
A high-performance liquid chromatography-tandem mass spectrometry (LC-MS-MS) method has been developed for the quantitative analysis of heroin and its major metabolites 6-acetylmorphine, morphine, morphine-3-glucuronide and morphine-6-glucuronide in blood and brain tissue, using 0.1-mL samples. We evaluated this method for analysis of heroin and its metabolites in samples from heroin treated mice. Ice-cold acidic buffer containing sodium fluoride was immediately added to blood and brain homogenate samples. Sample preparation was achieved by protein precipitation on ice-bath, using a mixture of ice-cold acetonitrile and methanol. The supernatant was evaporated to dryness, reconstituted with mobile phase, and injected into the chromatographic system. Separation was performed on a Xterra C18 column with gradient elution. The MS analysis was performed in positive ion mode, and multiple reaction monitoring (MRM) was used for drug quantification. The limits of quantification for the different opiates varied from 0.0007 to 0.02 mg/L in blood and from 0.002 to 0.06 microg/g in brain tissue. Day-to-day relative standard deviation ranged from 3.1 to 14.5%, and within-day variation ranged from 2.1 to 11.4%. The recoveries were between 80 and 111%. The stability of heroin was tested, and the study showed that heroin is more stable in brain tissue than in blood.
已开发出一种高效液相色谱-串联质谱（LC-MS-MS）方法，用于定量分析血液和脑组织中的海洛因及其主要代谢物6-乙酰吗啡、吗啡、吗啡-3-葡萄糖醛酸苷和吗啡-6-葡萄糖醛酸苷，使用的样本量为0.1 mL。我们评估了该方法用于分析海洛因处理小鼠样本中的海洛因及其代谢物。立即向血液和脑匀浆样本中加入含氟化钠的冰冷酸性缓冲液。通过在冰浴上用冰冷的乙腈和甲醇混合物进行蛋白质沉淀来制备样本。将上清液蒸发至干，用流动相复溶，然后注入色谱系统。在Xterra C18柱上进行梯度洗脱分离。质谱分析在正离子模式下进行，采用多反应监测（MRM）进行药物定量。不同阿片类药物在血液中的定量限为0.0007至0.02 mg/L，在脑组织中的定量限为0.002至0.06 μg/g。日间相对标准偏差范围为3.1%至14.5%，日内变化范围为2.1%至11.4%。回收率在80%至111%之间。对海洛因的稳定性进行了测试，研究表明海洛因在脑组织中比在血液中更稳定。
