High-performance liquid chromatographic assays for bufuralol 1'-hydroxylase, debrisoquine 4-hydroxylase, and dextromethorphan O-demethylase in microsomes and purified cytochrome P-450 isozymes of human liver
| ISSN: |
0003-2697
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| Keywords: |
HPLC ; bufuralol ; cytochrome P-450 ; debrisoquine, dextromethorphan ; drug metabolism ; human liver
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| Source: |
Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
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| Topics: |
Biology
Chemistry and Pharmacology
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| Type of Medium: |
Electronic Resource
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| URL: |
| _version_ | 1798292449652637696 |
|---|---|
| autor | Kronbach, T. Mathys, D. Gut, J. Catin, T. Meyer, U.A. |
| autorsonst | Kronbach, T. Mathys, D. Gut, J. Catin, T. Meyer, U.A. |
| book_url | http://dx.doi.org/10.1016/0003-2697(87)90006-6 |
| datenlieferant | nat_lic_papers |
| fussnote | Bufuralol, debrisoquine, and dextromethorphan are three prototype substrates of the common genetic deficiency of oxidative drug metabolism in man known as debrisoquine/sparteine-type polymorphism. We describe assays for the in vitro metabolism of (+)- and (-)-bufuralol, debrisoquine, and dextromethorphan in human liver microsomes and reconstituted purified cytochrome P-450 isozymes. These assays combine nonextractive sample preparation by precipitation of protein with perchloric acid with reversed-phase inorganic ion-pair HPLC and fluorescence detection. The minimal detectable levels of the major metabolites formed are 1'-hydroxybufuralol, 0.1 ng/ml; 4-hydroxydebrisoquine, 0.8 ng/ml; and dextrorphan, 0.1 ng/ml. Formation of these metabolites is linear for at least 45 min and between 1 and 100 μg of microsomal protein. Comparative kinetic analysis of the three monooxygenase reactions in human liver microsomes revealed an apparent biphasicity of (+)- and (-)-bufuralol 1'-hydroxylation and dextromethorphan O-demethylation but monophasic formation of 4-hydroxydebrisoquine in the substrate concentration range (〈1 mm) studied. These data, in combination with those obtained by purified human cytochrome P-450 isozymes indicate the involvement of the same enzyme in the metabolism of all three substrates investigated. However, additional and distinct activities contribute to the metabolism of (+)- and (-)-bufuralol and dextromethorphan. |
| hauptsatz | hsatz_simple |
| identnr | NLZ18406824X |
| issn | 0003-2697 |
| journal_name | Analytical Biochemistry |
| materialart | 1 |
| package_name | Elsevier |
| publikationsort | Amsterdam |
| publisher | Elsevier |
| reference | 162 (1987), S. 24-32 |
| schlagwort | HPLC bufuralol cytochrome P-450 debrisoquine, dextromethorphan drug metabolism human liver |
| search_space | articles |
| shingle_author_1 | Kronbach, T. Mathys, D. Gut, J. Catin, T. Meyer, U.A. |
| shingle_author_2 | Kronbach, T. Mathys, D. Gut, J. Catin, T. Meyer, U.A. |
| shingle_author_3 | Kronbach, T. Mathys, D. Gut, J. Catin, T. Meyer, U.A. |
| shingle_author_4 | Kronbach, T. Mathys, D. Gut, J. Catin, T. Meyer, U.A. |
| shingle_catch_all_1 | Kronbach, T. Mathys, D. Gut, J. Catin, T. Meyer, U.A. High-performance liquid chromatographic assays for bufuralol 1'-hydroxylase, debrisoquine 4-hydroxylase, and dextromethorphan O-demethylase in microsomes and purified cytochrome P-450 isozymes of human liver HPLC bufuralol cytochrome P-450 debrisoquine, dextromethorphan drug metabolism human liver HPLC bufuralol cytochrome P-450 debrisoquine, dextromethorphan drug metabolism human liver 0003-2697 00032697 Elsevier |
| shingle_catch_all_2 | Kronbach, T. Mathys, D. Gut, J. Catin, T. Meyer, U.A. High-performance liquid chromatographic assays for bufuralol 1'-hydroxylase, debrisoquine 4-hydroxylase, and dextromethorphan O-demethylase in microsomes and purified cytochrome P-450 isozymes of human liver HPLC bufuralol cytochrome P-450 debrisoquine, dextromethorphan drug metabolism human liver HPLC bufuralol cytochrome P-450 debrisoquine, dextromethorphan drug metabolism human liver 0003-2697 00032697 Elsevier |
| shingle_catch_all_3 | Kronbach, T. Mathys, D. Gut, J. Catin, T. Meyer, U.A. High-performance liquid chromatographic assays for bufuralol 1'-hydroxylase, debrisoquine 4-hydroxylase, and dextromethorphan O-demethylase in microsomes and purified cytochrome P-450 isozymes of human liver HPLC bufuralol cytochrome P-450 debrisoquine, dextromethorphan drug metabolism human liver HPLC bufuralol cytochrome P-450 debrisoquine, dextromethorphan drug metabolism human liver 0003-2697 00032697 Elsevier |
| shingle_catch_all_4 | Kronbach, T. Mathys, D. Gut, J. Catin, T. Meyer, U.A. High-performance liquid chromatographic assays for bufuralol 1'-hydroxylase, debrisoquine 4-hydroxylase, and dextromethorphan O-demethylase in microsomes and purified cytochrome P-450 isozymes of human liver HPLC bufuralol cytochrome P-450 debrisoquine, dextromethorphan drug metabolism human liver HPLC bufuralol cytochrome P-450 debrisoquine, dextromethorphan drug metabolism human liver 0003-2697 00032697 Elsevier |
| shingle_title_1 | High-performance liquid chromatographic assays for bufuralol 1'-hydroxylase, debrisoquine 4-hydroxylase, and dextromethorphan O-demethylase in microsomes and purified cytochrome P-450 isozymes of human liver |
| shingle_title_2 | High-performance liquid chromatographic assays for bufuralol 1'-hydroxylase, debrisoquine 4-hydroxylase, and dextromethorphan O-demethylase in microsomes and purified cytochrome P-450 isozymes of human liver |
| shingle_title_3 | High-performance liquid chromatographic assays for bufuralol 1'-hydroxylase, debrisoquine 4-hydroxylase, and dextromethorphan O-demethylase in microsomes and purified cytochrome P-450 isozymes of human liver |
| shingle_title_4 | High-performance liquid chromatographic assays for bufuralol 1'-hydroxylase, debrisoquine 4-hydroxylase, and dextromethorphan O-demethylase in microsomes and purified cytochrome P-450 isozymes of human liver |
| sigel_instance_filter | dkfz geomar wilbert ipn albert fhp |
| source_archive | Elsevier Journal Backfiles on ScienceDirect 1907 - 2002 |
| timestamp | 2024-05-06T08:48:41.155Z |
| titel | High-performance liquid chromatographic assays for bufuralol 1'-hydroxylase, debrisoquine 4-hydroxylase, and dextromethorphan O-demethylase in microsomes and purified cytochrome P-450 isozymes of human liver |
| titel_suche | High-performance liquid chromatographic assays for bufuralol 1'-hydroxylase, debrisoquine 4-hydroxylase, and dextromethorphan O-demethylase in microsomes and purified cytochrome P-450 isozymes of human liver Bufuralol, debrisoquine, and dextromethorphan are three prototype substrates of the common genetic deficiency of oxidative drug metabolism in man known as debrisoquine/sparteine-type polymorphism. We describe assays for the in vitro metabolism of (+)- and (-)-bufuralol, debrisoquine, and dextromethorphan in human liver microsomes and reconstituted purified cytochrome P-450 isozymes. These assays combine nonextractive sample preparation by precipitation of protein with perchloric acid with reversed-phase inorganic ion-pair HPLC and fluorescence detection. The minimal detectable levels of the major metabolites formed are 1'-hydroxybufuralol, 0.1 ng/ml; 4-hydroxydebrisoquine, 0.8 ng/ml; and dextrorphan, 0.1 ng/ml. Formation of these metabolites is linear for at least 45 min and between 1 and 100 μg of microsomal protein. Comparative kinetic analysis of the three monooxygenase reactions in human liver microsomes revealed an apparent biphasicity of (+)- and (-)-bufuralol 1'-hydroxylation and dextromethorphan O-demethylation but monophasic formation of 4-hydroxydebrisoquine in the substrate concentration range (〈1 mm) studied. These data, in combination with those obtained by purified human cytochrome P-450 isozymes indicate the involvement of the same enzyme in the metabolism of all three substrates investigated. However, additional and distinct activities contribute to the metabolism of (+)- and (-)-bufuralol and dextromethorphan. |
| topic | W V |
| uid | nat_lic_papers_NLZ18406824X |