A gas chromatography-mass spectrometry method for the measurement of fatty acid omega and omega(-1) hydroxylation kinetics by CYP4A1 using an artificial membrane system

Holmes, Victoria E., Bruce, Mary, Shaw, P. Nicholas, Bell, David R., Qi, Fan Ming and Barrett, David A. (2004) A gas chromatography-mass spectrometry method for the measurement of fatty acid omega and omega(-1) hydroxylation kinetics by CYP4A1 using an artificial membrane system. Analytical Biochemistry, 325 2: 354-363. doi:10.1016/j.ab.2003.10.046


Author Holmes, Victoria E.
Bruce, Mary
Shaw, P. Nicholas
Bell, David R.
Qi, Fan Ming
Barrett, David A.
Title A gas chromatography-mass spectrometry method for the measurement of fatty acid omega and omega(-1) hydroxylation kinetics by CYP4A1 using an artificial membrane system
Journal name Analytical Biochemistry   Check publisher's open access policy
ISSN 0003-2697
Publication date 2004-02-15
Year available 2004
Sub-type Article (original research)
DOI 10.1016/j.ab.2003.10.046
Volume 325
Issue 2
Start page 354
End page 363
Total pages 10
Place of publication San Diego
Publisher Academic Press Inc Elsevier Science
Language eng
Subject 0301 Analytical Chemistry
0601 Biochemistry and Cell Biology
Abstract A gas chromatography-mass spectrometry assay method for the analysis of lauric, myristic, and palmitic acids and their omega and omega-1 hydroxylated metabolites from in vitro incubations of cytochrome P450 CYP4A1, involving solid-phase extraction and trimethysityl derivatization, was developed. The assay was linear, precise, and accurate over the range 0.5 to 50 muM for all the analytes. It has the advantages of a more rapid analysis time, an improved sensitivity, and a wider range of analytes compared with other methods. An artificial membrane system was optimized for application to purified CYP4A1 enzyme by investigating the molar ratios of cytochrome b(5) and cytochrome P450 reductase present in the incubation mixture. Using this method, the kinetics of omega and omega(-1) oxidation of lauric, myristic, and palmitic acids by CYP4A enzymes were measured and compared in rat liver microsomes and an artificial membrane system. (C) 2003 Elsevier Inc. All rights reserved.
Keyword Biochemical Research Methods
Biochemistry & Molecular Biology
Chemistry, Analytical
fatty acids
hydroxylation
CYP4A1
artificial membrane
GC-MS
kinetics
Performance Liquid-chromatography
Lauric-acid
Rat-liver
P450 2e1
Cytochrome-p450
Protein
Regiospecificity
Microsomes
Clofibrate
Induction
Q-Index Code C1

Document type: Journal Article
Sub-type: Article (original research)
Collections: Excellence in Research Australia (ERA) - Collection
School of Pharmacy Publications
 
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