title: An HPLC assay for sn-1,2-diacylglycerol.
abstract: A method for the analysis of 1,2-diacylglycerols in biological samples is presented. After tissue extraction and derivatisation with 3,5-dinitrobenzoyl chloride, samples are analysed by normal phase HPLC, using a 3.9 x 300 mm microPorasil column, and ultraviolet detection at 254 nm. The method gives quantitative recovery of 1,2-diacylglycerol, and is of sufficient sensitivity to allow quantitation of 1,2-diacylglycerol in human muscle needle biopsy specimens, from as little as 10 mg muscle. Human skeletal muscle from fasted control subjects was found to have a 1,2-diacylglycerol content of 455 +/- 78 nmol/g wet weight. The method is robust, giving intra- and inter-assay coefficients of variation of 2.9% and 5.9%, respectively, and should prove useful for the analysis of 1,2-diacylglycerol levels in human disease states, such as diabetes, in which no measurements of 1,2-diacylglycerol have yet been undertaken.
authors.affiliation: Department of Medicine, Medical School, University of Newcastle upon Tyne, UK.
authors.forename: S
authors.initials: S
authors.lastname: Eaton
authors.forename: E
authors.initials: E
authors.lastname: Shmueli
authors.forename: H
authors.initials: H
authors.lastname: al-Mardini
authors.forename: K
authors.initials: K
authors.lastname: Bartlett
authors.forename: C O
authors.initials: CO
authors.lastname: Record
mesh_terms: D000818:Animals; D002851:Chromatography, High Pressure Liquid; D004075:Diglycerides / Q000032:analysis*; D006801:Humans; D008297:Male; D018482:Muscle, Skeletal / Q000737:chemistry; D051381:Rats; D017208:Rats, Wistar; D013056:Spectrophotometry, Ultraviolet
chemical_list: C011439:1,2-diacylglycerol; D004075:Diglycerides