title: A rapid procedure for the separation and analysis of metabolites of the sterol and dolichol pathways.
abstract: We have previously shown that of several methods examined, the maximum yield of dolichyl phosphate (Dol-P) from rat liver was achieved by saponification of the tissue and subsequent extraction with diethyl ether (R.K. Keller et al. (1985) Anal. Biochem. 147, 166-173). In the present report, we have developed a rapid procedure using non-toxic solvents which resolves the ether extract on a C18 cartridge column into four major fractions: (1) fatty acids; (2) squalene and sterols; (3) dolichol; (4) Dol-P. The utility of the new procedure was demonstrated by preparing the four fractions from liver slices which had been incubated with [3H]acetate. HPLC analysis of the sterol, dolichol and Dol-P fractions yielded well resolved elution profiles, thereby allowing determination of radioactivity incorporated into the major isoprenoids and their metabolites.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of South Florida, College of Medicine, Tampa 33612.
authors.forename: R K
authors.initials: RK
authors.lastname: Keller
authors.forename: S W
authors.initials: SW
authors.lastname: Nellis
authors.forename: L
authors.initials: L
authors.lastname: Cuadrado-Simonet
mesh_terms: D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004288:Dolichol Phosphates / Q000302:isolation & purification / Q000378:metabolism; D004286:Dolichols / Q000302:isolation & purification* / Q000378:metabolism; D066298:In Vitro Techniques; D008099:Liver / Q000378:metabolism; D008297:Male; D051381:Rats; D011919:Rats, Inbred Strains; D012997:Solvents; D013261:Sterols / Q000302:isolation & purification* / Q000378:metabolism
chemical_list: D004288:Dolichol Phosphates; D004286:Dolichols; D012997:Solvents; D013261:Sterols