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Monolithic silica rod liquid chromatography with ultraviolet or fluorescence detection for metabolite analysis of cytochrome P450 marker reactions
DMPK & Bioanalytical Chemistry, AstraZeneca R&D Mölndal, Mölndal, Sweden.
DMPK & Bioanalytical Chemistry, AstraZeneca R&D Mölndal, Mölndal, Sweden.
DMPK & Bioanalytical Chemistry, AstraZeneca R&D Mölndal, Mölndal, Sweden.
2002 (English)In: Journal of chromatography. B, ISSN 1570-0232, E-ISSN 1873-376X, Vol. 780, no 2, 205-215 p.Article in journal (Refereed) Published
Abstract [en]

In vitro cytochrome P450 assays are used in metabolism studies in support of early phases of drug discovery to investigate, e.g., metabolic stability, enzyme inhibition and induction by new chemical entities. LC-UV and LC-fluorescence are traditional analytical tools in support of such studies. However, these tools typically comprise different methods of relatively low throughput for the various metabolites of probe reactions. In recent years, LC-MS methods have been developed to increase throughput. Increased throughput can also be achieved by means of modern chromatographic tools in combination with UV and fluorescence detection. This approach is especially suitable when cytochrome P450 isoforms are investigated by means of single probe incubations. Here, an LC-UV/fluorescence system based on a monolithic porous silica column is described for the analysis of metabolites of nine cytochrome P450 marker reactions [phenacetin to paracetamol (CYP1A2), coumarin to 7-hydroxycoumarin (CYP2A6), paclitaxel to 6alpha-hydroxypaclitaxel (CYP2C8), diclofenac to 4-hydroxydiclofenac (CYP2C9), mephenytoin to 4-hydroxymephenytoin (CYP2C19), bufuralol to 1-hydroxybufuralol (CYP2D6), chlorzoxazone to 6-hydroxychlorzoxazone (CYP2E1), midazolam to 1-hydroxymidazolam (CYP3A4), and testosteron to 6beta-hydroxytestosteron (CYP3A4)]. While offering sensitivities and linear ranges comparable to previously reported methods, the set-up described here provides ease of use and increased throughput with maximum cycle times of 4.5 min. © 2002 Elsevier Science B.V. All rights reserved.

Place, publisher, year, edition, pages
Amsterdam: Elsevier, 2002. Vol. 780, no 2, 205-215 p.
Keyword [en]
Monolithic silica rod, Cytochrome P450
National Category
Analytical Chemistry
Identifiers
URN: urn:nbn:se:hh:diva-24120DOI: 10.1016/S1570-0232(02)00264-7ISI: 000179132000001PubMedID: 12401345Scopus ID: 2-s2.0-0037175432OAI: oai:DiVA.org:hh-24120DiVA: diva2:679540
Available from: 2013-12-16 Created: 2013-12-09 Last updated: 2015-03-03Bibliographically approved

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