Analysis of epoxyeicosatrienoic acids by chiral liquid chromatography/electron capture atmospheric pressure chemical ionization mass spectrometry using [13C]-analog internal standards.
Analysis of epoxyeicosatrienoic acids by chiral liquid chromatography/electron capture atmospheric pressure chemical ionization mass spectrometry using [13C]-analog internal standards.
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DOI：
10.1002/rcm.4760
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发表时间：
2010-11-30
影响因子：
2
通讯作者：
Blair, Ian A.
中科院分区：
文献类型：
--
作者：
Mesaros, Clementina;Lee, Seon Hwa;Blair, Ian A.
花生四烯酸（AA）向环氧二碳三烯酸（EETs）的代谢被认为主要是由来自2家族（2C9, 2C19, 2D6和2J2）的细胞色素P450 （P450）介导的。相反，4家族的p450主要参与AA的omega氧化（4A11和4A22）。确定区域异构体eet的对映选择性形成的能力对于确定其潜在的生物活性和评估哪些P450同工异构体参与其形成非常重要。分析生物液体中的EET对映体非常困难，因为它们仅以微量存在，而且它们彼此分离非常困难。此外，通常用于稳定同位素稀释液相色谱/质谱（LC/MS）分析的氘标记内标物与相应的protium形式相比具有不同的LC保留时间。因此，基于LC/ ms的定量方法可能会受到紧密洗脱异构体电离的差异抑制。我们报道了[13C20]-EET模拟内标的制备，并使用经过验证的高灵敏度手性LC/电子捕获大气压化学电离(ECAPCI)-MS方法对内源性eet作为其五氟苯（PFB）酯衍生物进行痕量分析。随后，该实验显示了P4502C19-、P4502D6-、P4501A1-和p4501b1介导的AA转化为eet的精细对映选择性，并量化了小鼠上皮性肝癌（Hepa）细胞系中AA代谢产生的eet的对映选择性。
The metabolism of arachidonic acid (AA) to epoxyeicosatrienoic acids (EETs) is thought to be mediated primarily by the cytochromes P450 (P450s) from the 2 family (2C9, 2C19, 2D6, and 2J2). In contrast, P450s of the 4 family are primarily involved in omega oxidation of AA (4A11 and 4A22). The ability to determine enantioselective formation of the regioisomeric EETs is important in order to establish their potential biological activities and to asses which P450 isoforms are involved in their formation. It has been extremely difficult to analyze individual EET enantiomers in biological fluids because they are present in only trace amounts and they are extremely difficult to separate from each other. In addition, the deuterium-labeled internal standards that are commonly used for stable isotope dilution liquid chromatography/mass spectrometry (LC/MS) analyses have different LC retention times when compared with the corresponding protium forms. Therefore, quantification by LC/MS-based methodology can be compromised by differential suppression of ionization of the closely eluting isomers. We report the preparation of [13C20]-EET analog internal standards and the use of a validated high-sensitivity chiral LC/electron capture atmospheric pressure chemical ionization (ECAPCI)-MS method for the trace analysis of endogenous EETs as their pentafluorobenzyl (PFB) ester derivatives. The assay was then used to show the exquisite enantioselectivity of P4502C19-, P4502D6-, P4501A1-, and P4501B1-mediated conversion of AA into EETs and to quantify the enantioselective formation of EETs produced by AA metabolism in a mouse epithelial hepatoma (Hepa) cell line.
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DOI：
10.1016/0006-291x(87)90576-6
发表时间：
1987-07-31
影响因子：
3.1
作者：
CAPDEVILA, JH;KISHORE, V;FALCK, JR
通讯作者：
FALCK, JR
DOI：
10.1073/pnas.85.8.2528
发表时间：
1988-04-01
影响因子：
11.1
作者：
DENISON, MS;FISHER, JM;WHITLOCK, JP
通讯作者：
WHITLOCK, JP
影响因子：
4.8
作者：
CAPDEVILA, JH;FALCK, JR;ESTABROOK, RW
通讯作者：
ESTABROOK, RW
影响因子：
--
作者：
KATOH, T;TAKAHASHI, K;BADR, KF
通讯作者：
BADR, KF
影响因子：
3.9
作者：
CHACOS, N;CAPDEVILA, J;ESTABROOK, RW
通讯作者：
ESTABROOK, RW