HPLC-MS/MS analysis of the products generated from all-trans-retinoic acid using recombinant human CYP26A
HPLC-MS/MS analysis of the products generated from all-trans-retinoic acid using recombinant human CYP26A
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DOI：
10.1194/jlr.m100343-jlr200
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发表时间：
2002-07-01
影响因子：
6.5
通讯作者：
Jones, G
中科院分区：
文献类型：
--
作者：
Chithalen, JV;Luu, L;Jones, G
两个哺乳动物hCYP26A表达系统被用来分析CYP26A的代谢产物。通过广泛的高效液相色谱、紫外光谱和液相色谱/串联质谱（LC-MS/MS）方法，我们最终证明了复杂的混合物包括4- oh -全反式维甲酸、4-氧-全反式维甲酸和18- oh -全反式维甲酸，以及更多的极性产物，部分被鉴定为二羟基和单氧、单羟基衍生物。这些极性更强的产物被认为是β -离子酮环上多次羟基化的结果。初始代谢物和极性代谢物的相互关系从基因剂量和时间过程实验中推断出来。4-氧-全反式维甲酸后的初始和次级代谢步骤都是酮康唑敏感的，这表明水溶性代谢物的产生步骤依赖于细胞色素p450。利用重组人CYP26A对全反式维甲酸产物进行HPLC-MS/MS分析。
Two mammalian hCYP26A expression systems have been used to analyze the metabolic products of CYP26A. Through the use of extensive HPLC, UV spectroscopy, and liquid chromatography/tandem mass spectrometry (LC-MS/MS) methodology, we have conclusively demonstrated that the complex mixture of products comprises 4-OH-all-trans-retinoic acid, 4-oxo-all-trans-retinoic acid, and 18-OH-all-trans-retinoic acid, and more polar products, partially identified as dihydroxy and mono-oxo, mono-hydroxy derivatives. These more polar products are presumed to result from multiple hydroxylations on the beta-ionone ring. The inter-relationship of initial and polar metabolites was inferred from both gene-dose and time-course experiments. Both initial and secondary metabolic steps after 4-oxo-all-trans-retinoic acid are ketoconazole-sensitive, suggesting that steps in the production of water-soluble metabolites are cytochrome P450-dependent. HPLC-MS/MS analysis of the products generated from all-trans-retinoic acid using recombinant human CYP26A.