Separation of the apoprotein components of human very low density lipoproteins by ion-paired, reversed-phase high performance liquid chromatography.
A number of crude apolipoprotein samples isolated from human very low density lipoproteins (VLDL) were analyzed by reversed phase high performance liquid chromatography. The mobile phase consisted of a 1% solution of the polar ion-pairing reagent triethylammonium phosphate. A slow, nonlinear gradient of acetonitrile (37--42%) was used to elute the apolipoproteins. The order of elution was as follows: apolipoprotein CX, apolipoprotein C-I, apolipoprotein C-III2, apolipoprotein C-III1, apolipoprotein C-IIIQ and apolipoprotein C-II. This order is consistent with the known polarity of the proteins, i.e., the most nonpolar, apolipoprotein C-II, was the last to be eluted, whereas apolipoprotein C-I, with the lowest nonpolar surface area eluted first. The recovery of the individual apolipoproteins was 80--95% and the individual peaks were characterized by amino acid analysis, UV absorption spectra amd chromatography of pure protein standards.
D000097:Acetonitriles; D053302:Apolipoprotein C-I; D053304:Apolipoprotein C-II; D053305:Apolipoprotein C-III; D001053:Apolipoproteins / Q000302:isolation & purification*; D001056:Apolipoproteins C; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D005021:Ethylamines; D006801:Humans; D008079:Lipoproteins, VLDL / Q000097:blood*
D000097:Acetonitriles; D053302:Apolipoprotein C-I; D053304:Apolipoprotein C-II; D053305:Apolipoprotein C-III; D001053:Apolipoproteins; D001056:Apolipoproteins C; D005021:Ethylamines; D008079:Lipoproteins, VLDL; C016162:triethylamine