High-performance liquid chromatographic analysis of acylated lipids containing pyrene fatty acids.
A high-performance liquid chromatographic method for the separation and quantification of acylated lipids containing pyrene fatty acids is described. The method is adapted from a procedure originally developed for the analysis of tissue lipids (Christie, W. W. (1985) J. Lipid Res. 26, 507-512). Pyrenyl lipid analogs ranging in polarity from cholesteryl ester to lysophosphatidylcholine are completely resolved on a silica column in 50 min by gradient elution with a ternary solvent system. Furthermore, pyrene-labeled triglycerides are resolved according to the number of pyrene fatty acid residues incorporated. Pyrenyl lipids are detected at levels of 10(-13) mol by high-sensitivity fluorescence detection. Accurate quantification of pyrenyl lipids is obtained by correcting peak areas for mobile-phase quenching effects. The close correspondence between chromatograms obtained for the separation of labeled lipids extracted from Hep-G2 cells incubated with either 12-(1-pyrenyl)dodecanoic acid (fluorescence detection) or [1-14C]oleic acid (radioactivity detection) indicates that this HPLC method is equally suitable for analysis of native lipids.
Homan
R
R
Baylor College of Medicine, Houston, Texas 77030.
Pownall
H J
HJ
D006528:Carcinoma, Hepatocellular / Q000032:analysis; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D005227:Fatty Acids / Q000032:analysis*; D008055:Lipids / Q000032:analysis / Q000302:isolation & purification*; D008113:Liver Neoplasms; D011721:Pyrenes / Q000032:analysis*
D005227:Fatty Acids; D008055:Lipids; D011721:Pyrenes