Development of miniaturized drug metabolism system based on capillary electromigration methods
| Authors | |
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| Year of publication | 2007 |
| Type | Article in Proceedings |
| Conference | HPLC 2007. Abstract Book. |
| MU Faculty or unit | |
| Citation | |
| Web | 31st International Symposium on High Performance Liquid Phase Separations and Related Techniques - HPLC 2007 |
| Field | Analytic chemistry |
| Keywords | CE; drug metabolism |
| Description | The study of drug and xenobiotic metabolism is essential for understanding of targeted disease treatment. In particular high performance liquid chromatography (HPLC) with UV and fluorescence detection is predominantly chosen for studies. The standard methods often consume large sample quantities which is a disadvantage in investigation of enzymatic systems. Examination of drug metabolism and interactions can also be performed on capillary electrophoresis (CE) equipment. Because of sample saving and minor time demands, CE analysis is suitable for studies comprising work with enzymes, substrates as well as inhibitors. The automation of all assay steps also contributes to elimination of experimental errors. The advantage in comparison with HPLC determination is the possibility to avoid the extraction step during sample preparation. Cytochromes P450 (CYP450s) are involved in the metabolism of a wide variety of xenobiotic chemicals. One major role is the clearance of drug, to which CYP450s collectively contribute more than any other group of enzymes. These enzymes are a large group of membrane bound hemoproteins. They exhibit a large spectrum of substrate specifity and can exist in various forms, called isoforms. The mammalian CYP450s take apart in biosynthesis and metabolism of sterols and steroid hormones and in metabolism of various lipoidic biofactors. The main function of cytochromes P450 is the oxidative metabolism of xenobiotics. The xenobiotic metabolism in the human body was studied with the use of microsomes, as a source of CYP450 enzymes, and dextromethorphan, as a substrate. The most involved CYP450 isoforms 3A4 and 2D6 transform the substrate to 3-methoxymorphinan, 3-hydroxymorphinan and dextrorphan. A background electrolyte consisting of borate buffer (75mM, pH 9.8) allows to separate these four compounds. To monitor the drug metabolism the incubation with microsomes was performed. |
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