Conventional liquid chromatography/triple quadrupole mass spectrometry based metabolite identification and semi-quantitative estimation approach in the investigation of in vitro dabigatran etexilate metabolism.
Conventional liquid chromatography/triple quadrupole mass spectrometry based metabolite identification and semi-quantitative estimation approach in the investigation of in vitro dabigatran etexilate metabolism.
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在体外dabigatran et催化代谢的研究中，常规的液相色谱/三极质质谱法代谢物鉴定和半定量估计方法。
DOI：
10.1007/s00216-012-6576-4
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发表时间：
2013-02
影响因子：
4.3
通讯作者：
Laizure, S. Casey
中科院分区：
文献类型：
--
作者：
Hu, Zhe-Yi;Parker, Robert B.;Herring, Vanessa L.;Laizure, S. Casey
达比加群酯（DABE）是一种口服前药，可通过酯酶快速转化为达比加群（DAB），后者是凝血酶的直接抑制剂。为了阐明酯酶介导的DABE代谢途径，建立了基于高效液相色谱/质谱（LC-MS/MS）的代谢物鉴定和半定量估计方法。为克服传统三重四极杆质谱全扫描灵敏度差的缺点，采用预测子-产物离子对的方法，寻找潜在的体外代谢产物。通过产物离子扫描确认检测到的代谢物。引入稀释法来评价在没有化学标准品的情况下初步鉴定的代谢物的基质效应。使用由含有高浓度代谢物的孵育样品生成的“代谢物标准品”结合质谱响应的校正因子，获得了检测到的代谢物的定量信息。鉴定了DABE的两种体外代谢产物（M1和M2），并通过半定量估计方法进行了定量。值得注意的是，CES 1将DABE转换为M1，而CES 2介导DABE转换为M2。M1（或M2）通过CES 2（或CES 1）进一步代谢为DAB。这里提出的方法提供了一个解决方案，以快速识别和半定量估计临床前样品中的CES代谢物的生物分析需要。
Dabigatran etexilate (DABE) is an oral prodrug that is rapidly converted by esterases to dabigatran (DAB), a direct inhibitor of thrombin. To elucidate the esterase-mediated metabolic pathway of DABE, a high-performance liquid chromatography/mass spectrometer (LC-MS/MS)-based metabolite identification and semi-quantitative estimation approach was developed. To overcome the poor full-scan sensitivity of conventional triple quadrupole mass spectrometry, precursor-product ion pairs were predicted, to search for the potential in vitro metabolites. The detected metabolites were confirmed by the product ion scan. A dilution method was introduced to evaluate the matrix effects of tentatively identified metabolites without chemical standards. Quantitative information on detected metabolites was obtained using ‘metabolite standards’ generated from incubation samples that contain a high concentration of metabolite in combination with a correction factor for mass spectrometry response. Two in vitro metabolites of DABE (M1 and M2) were identified, and quantified by the semi-quantitative estimation approach. It is noteworthy that CES1 convert DABE to M1 while CES2 mediates the conversion of DABE to M2. M1 (or M2) was further metabolized to DAB by CES2 (or CES1). The approach presented here provides a solution to a bioanalytical need for fast identification and semi-quantitative estimation of CES metabolites in preclinical samples.
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DOI：
10.1016/j.jpba.2011.09.018
发表时间：
2012-01-25
影响因子：
3.4
作者：
Delavenne, Xavier;Moracchini, Julie;Basset, Thierry
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