UPLC-MS/MS方法研究次乌头碱在大鼠肝微粒体CYP450中的体外代谢产物及代谢酶亚型

毕云枫, 李雪, 皮子凤, 宋凤瑞, 刘志强

毕云枫, 李雪, 皮子凤, 宋凤瑞, 刘志强. UPLC-MS/MS方法研究次乌头碱在大鼠肝微粒体CYP450中的体外代谢产物及代谢酶亚型[J]. 质谱学报, 2013, 34(6): 330-337. DOI: 10.7538/zpxb.2013.34.06.0330
引用本文: 毕云枫, 李雪, 皮子凤, 宋凤瑞, 刘志强. UPLC-MS/MS方法研究次乌头碱在大鼠肝微粒体CYP450中的体外代谢产物及代谢酶亚型[J]. 质谱学报, 2013, 34(6): 330-337. DOI: 10.7538/zpxb.2013.34.06.0330
BI Yun-feng, LI Xue, PI Zi-feng, SONG Feng-rui, LIU Zhi-qiang. Analysis of Hypaconitine’s Metabolites and Related Metabolic CYP Isoforms in Rat Liver Microsomal by UPLC-MS/MS[J]. Journal of Chinese Mass Spectrometry Society, 2013, 34(6): 330-337. DOI: 10.7538/zpxb.2013.34.06.0330
Citation: BI Yun-feng, LI Xue, PI Zi-feng, SONG Feng-rui, LIU Zhi-qiang. Analysis of Hypaconitine’s Metabolites and Related Metabolic CYP Isoforms in Rat Liver Microsomal by UPLC-MS/MS[J]. Journal of Chinese Mass Spectrometry Society, 2013, 34(6): 330-337. DOI: 10.7538/zpxb.2013.34.06.0330
毕云枫, 李雪, 皮子凤, 宋凤瑞, 刘志强. UPLC-MS/MS方法研究次乌头碱在大鼠肝微粒体CYP450中的体外代谢产物及代谢酶亚型[J]. 质谱学报, 2013, 34(6): 330-337. CSTR: 32365.14.zpxb.2013.34.06.0330
引用本文: 毕云枫, 李雪, 皮子凤, 宋凤瑞, 刘志强. UPLC-MS/MS方法研究次乌头碱在大鼠肝微粒体CYP450中的体外代谢产物及代谢酶亚型[J]. 质谱学报, 2013, 34(6): 330-337. CSTR: 32365.14.zpxb.2013.34.06.0330
BI Yun-feng, LI Xue, PI Zi-feng, SONG Feng-rui, LIU Zhi-qiang. Analysis of Hypaconitine’s Metabolites and Related Metabolic CYP Isoforms in Rat Liver Microsomal by UPLC-MS/MS[J]. Journal of Chinese Mass Spectrometry Society, 2013, 34(6): 330-337. CSTR: 32365.14.zpxb.2013.34.06.0330
Citation: BI Yun-feng, LI Xue, PI Zi-feng, SONG Feng-rui, LIU Zhi-qiang. Analysis of Hypaconitine’s Metabolites and Related Metabolic CYP Isoforms in Rat Liver Microsomal by UPLC-MS/MS[J]. Journal of Chinese Mass Spectrometry Society, 2013, 34(6): 330-337. CSTR: 32365.14.zpxb.2013.34.06.0330

UPLC-MS/MS方法研究次乌头碱在大鼠肝微粒体CYP450中的体外代谢产物及代谢酶亚型

Analysis of Hypaconitine’s Metabolites and Related Metabolic CYP Isoforms in Rat Liver Microsomal by UPLC-MS/MS

  • 摘要: 次乌头碱是一种二萜类双酯型乌头碱类化合物,广泛存在于乌头属植物中。本工作利用超高效液相色谱-串联质谱联用(UPLC-MS/MS)及高分辨质谱(HRMS)法分析确定了次乌头碱在大鼠肝微粒体CYP450中的代谢产物,分别选取磺胺苯吡唑、α-萘黄酮、奎那定、酮康唑、二乙基二硫代氨基甲酸铵作为CYP2C、CYP1A2、CYP2D、CYP3A、CYP2E1的特异性抑制剂,确定了各产物的CYP450酶的代谢亚型。结果表明:次乌头碱在大鼠肝微粒体中的主要代谢产物有7种,分别为15-脱氢次乌头碱、8-O-脱乙酰基-次乌头碱、2-羟基-次乌头碱、中乌头碱、1-去甲基-次乌头碱、18-去甲基-次乌头碱和羟基-8-O-脱乙酰基-次乌头碱;CYP3A为次乌头碱的主要代谢酶,CYP2C、CYP1A2、CYP2D和CYP2E1也参与了次乌头碱的代谢。
    Abstract: Hypaconitine (HA) is a diester-diterpene type aconitum alkaloid, a potential toxicity alkaloid extracted from Aconitum longtounense T. In this study, the hypaconitine’s metabolites in rat liver microsomes were determined by UPLC-MS/MS and a high resolution mass spectrogram (HRMS) method. Sulfaphenazole, α-naphthoflavone, quinidine, ketoconazole, ammonium diethyldithiocarbamate were applied as specific inhibitors for CYP3A、CYP1A2、CYP2D6、CYP3A and CYP2E1 in rat liver microsomes. The results show that seven metabolites of hypaconitine were found and characterized in rat liver microsomal incubations. These metabolites were deduced as 15-dehydrate-HA, 8-O-deacetyl-HA, 2-hydroxyl-HA, mesaconitine, 1-O-demethyl-HA, 18-O-demethyl-HA, hydroxyl-8-deacetyl-HA, respectively. Hypaconitine was mainly metabolized by CYP3A. CYP2C, CYP1A2,CYP2D and CYP2E1 were important CYP isoforms responsible for the metabolism reaction of hypaconitine.
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