Qualitative analysis of zolpidem and its metabolites M-1 to M-4 in human blood and urine using liquid chromatography-tandem mass spectrometry
Qualitative analysis of zolpidem and its metabolites M-1 to M-4 in human blood and urine using liquid chromatography-tandem mass spectrometry
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DOI：
10.1007/s11419-020-00551-w
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发表时间：
2020-09-17
影响因子：
2.2
通讯作者：
Kanawaku, Yoshimasa
中科院分区：
文献类型：
--
作者：
Yamaguchi, Koji;Miyaguchi, Hajime;Kanawaku, Yoshimasa
目的建立人血液和尿液中唑吡坦及其代谢产物M-1 ~ M-4的液相色谱-串联质谱（LC-MS/MS）定性分析方法。方法建立唑吡坦代谢物M-1 ~ M-4的合成方法，获得分析标准品。采用铜催化的三组分偶联反应和吡咯烷催化的亚胺生成反应相结合的方法合成了M-1 （M-1- me）和M-2 （M-2- me）甲酯，分别水解得到M-1和M-2。同时合成了M-3和M-4。采用标准品对LC-MS/MS条件进行优化。在人的血液和尿液中分析M-1至M-4。结果成功合成了M-1 ~ M-4。M-1和M-2的质谱几乎相同，但它们的峰是用十八烷基硅柱色谱分离的。同样，M-3和M-4的质谱相似，但它们的峰是色谱分离的。人体血液和尿液中代谢物的峰值强度为M-1 > M-2和M-4 > M-3；M-3仅在尿液中检测到。还发现了其他羟唑吡坦的存在。结论M-1和M-2以及M-4等羟基唑吡坦的质谱相似，在分析生物基质中唑吡坦代谢物时需要进行色谱分离。本研究首次采用LC-MS/MS同时分析唑吡坦和M-1至M-4，为唑吡坦的摄入提供更有说服力的证据。
Purpose The aim of this study was to develop the method for the qualitative analysis of zolpidem and its metabolites M-1 to M-4 using liquid chromatography-tandem mass spectrometry (LC-MS/MS) in human blood and urine. Methods To obtain analytical standards, a method for synthesizing zolpidem metabolites M-1 to M-4 was developed. A combination of a copper-catalyzed three-component coupling reaction and pyrrolidine-catalyzed imine formation was adopted to synthesize the methyl esters of M-1 (M-1-Me) and M-2 (M-2-Me), which were hydrolyzed to afford M-1 and M-2, respectively. M-3 and M-4 were also synthesized. The LC-MS/MS conditions were optimized using authentic standards. M-1 to M-4 were analyzed in human blood and urine. Results M-1 to M-4 were successfully synthesized. The mass spectra of M-1 and M-2 were almost identical, but their peaks were chromatographically separated using a octadecyl silica column. Likewise, mass spectra of M-3 and M-4 were similar, but their peaks were chromatographically separated. The peak intensities of the metabolites in human blood and urine were M-1 > M-2, and M-4 > M-3; M-3 was detected only in urine. The presence of other hydroxyzolpidems was also revealed. Conclusions Chromatographic separation of M-1 and M-2 and that of M-4 and other hydroxyzolpidems is necessary to analyze zolpidem metabolites in biological matrices because their mass spectra are quite similar. This study is the first to analyze zolpidem and M-1 to M-4 simultaneously using LC-MS/MS, which can provide more compelling evidence of zolpidem intake.