Evaluation of 3-hydroxy-3-methylglutaryl-COA-reductase, cholesterol-7α-hydroxylase and acyl-COA:cholesterol acyltransferase activities: alternative chromatographic methods to separate metabolites

Alternative HPLC and solid‐phase extraction column methods were developed to separate metabolites of enzymes involved in cholesterol metabolism in rabbit liver microsomes: hydroxyl‐methylglutaryl‐CoA reductase, cholesterol‐7α‐hydroxylase and acyl‐CoA:cholesterol acyltransferase. A comparison method...

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Bibliographic Details
Published in:Biomedical chromatography 2004-11, Vol.18 (9), p.706-713
Main Authors: Montoudis, Alain, Boileau, Serge, Simoneau, Lucie, Mounier, Catherine, Lafond, Julie
Format: Article
Language:English
Subjects:
ACAT
cholesterol-7α-hydroxylase
hepatic microsomes
HMG-CoA reductase
HPLC
SPE column
TLC plate
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Summary:Alternative HPLC and solid‐phase extraction column methods were developed to separate metabolites of enzymes involved in cholesterol metabolism in rabbit liver microsomes: hydroxyl‐methylglutaryl‐CoA reductase, cholesterol‐7α‐hydroxylase and acyl‐CoA:cholesterol acyltransferase. A comparison method of thin‐layer chromatography and solid‐phase extraction column were assayed to separate substrate and metabolite of hydroxy‐methylglutaryl‐CoA reductase, whereas for cholesterol‐7α‐hydroxylase and acyl‐CoA:cholesterol acyltransferase, this comparison was done between thin layer chromatography and HPLC. The results obtained by the new analytical chromatographic methods are not significantly different than those observed in literature. Moreover a larger percentage recovery was obtained for analysed metabolites. Our results demonstrate the reliability of these alternative chromatographic techniques and showed that they are valuable tools to precisely and rapidly measure the activity of those enzymes. Copyright © 2004 John Wiley & Sons, Ltd.
ISSN:0269-3879
1099-0801
DOI:10.1002/bmc.381