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Evaluation of 3‐hydroxy‐3‐methylglutaryl‐COA‐reductase, cholesterol‐7 α ‐hydroxylase and acyl‐COA:cholesterol acyltransferase activities: alternative chromatographic methods to separate metabolites
Author(s) -
Montoudis Alain,
Boileau Serge,
Simoneau Lucie,
Mounier Catherine,
Lafond Julie
Publication year - 2004
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.381
Subject(s) - chemistry , chromatography , reductase , sterol o acyltransferase , metabolite , cholesterol , enzyme , acyl coa , high performance liquid chromatography , biochemistry , lipoprotein
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‐7 α ‐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‐7 α ‐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 signicantly 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.