J. Lipid Res.
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Journal of Lipid Research, Vol 30, 454-457, Copyright © 1989 by Lipid Research, Inc.

ARTICLES

Quantitative analysis of retinal glycerolipid molecular species acetylated by acetolysis

HG Choe, RD Wiegand and RE Anderson
Program in Neuroscience, Baylor College of Medicine, Houston, TX 77030.

A method for the quantitative analysis of molecular species of 1,2- diacylglycerol acetates (1,2-DGAC) containing polyunsaturated fatty acids is described. Phosphatidylethanolamine (PE) isolated from frog retina was used to test the method. PE was converted to 1,2-DGAC by acetolysis. The molecular species of the 1,2-DGAC were resolved by reverse-phase high performance liquid chromatography (HPLC), detected by UV absorption spectroscopy at 210 nm, and identified by gas-liquid chromatography (GLC) of the fatty acid methyl esters (FAME). Molar response curves were generated for each DGAC molecular species that eluted as a single entity from HPLC by determining the moles of fatty acids in the molecular species collected and the response (peak area unit) of the UV detector. Each molecular species response curve was linear from about 10 pmoles to 4-8 nmoles, allowing the slope of each curve to be used as a molar absorptivity. This method provides a means for quantification of most of the molecular species of all glycerolipid classes.
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Copyright © 1989 by the American Society for Biochemistry and Molecular Biology.