title: Evaluation of 3-hydroxy-3-methylglutaryl-COA-reductase, cholesterol-7alpha-hydroxylase and acyl-COA:cholesterol acyltransferase activities: alternative chromatographic methods to separate metabolites.
abstract: 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-7alpha-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-7alpha-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.
authors.affiliation: Laboratoire de Physiologie Materno-Foetale, Université du Québec à Montréal, Montréal, Canada H3C 3P8.
authors.forename: Alain
authors.initials: A
authors.lastname: Montoudis
authors.forename: Serge
authors.initials: S
authors.lastname: Boileau
authors.forename: Lucie
authors.initials: L
authors.lastname: Simoneau
authors.forename: Catherine
authors.initials: C
authors.lastname: Mounier
authors.forename: Julie
authors.initials: J
authors.lastname: Lafond
mesh_terms: D000818:Animals; D002790:Cholesterol 7-alpha-Hydroxylase / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006903:Hydroxymethylglutaryl CoA Reductases / Q000378:metabolism*; D008297:Male; D011817:Rabbits; D013056:Spectrophotometry, Ultraviolet; D002785:Sterol O-Acyltransferase / Q000378:metabolism*
chemical_list: D006903:Hydroxymethylglutaryl CoA Reductases; D002790:Cholesterol 7-alpha-Hydroxylase; D002785:Sterol O-Acyltransferase