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Separation of the apoprotein components of human very low density lipoproteins by ion‐paired, reversed‐phase high performance liquid chromatography
Author(s) -
Hancock W. S.,
Bishop C. A.,
Gotto A. M.,
Harding D. R. K.,
Lamplugh S. M.,
Sparrow J. T.
Publication year - 1981
Publication title -
lipids
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.601
H-Index - 120
eISSN - 1558-9307
pISSN - 0024-4201
DOI - 10.1007/bf02535025
Subject(s) - lipidology , chromatography , clinical chemistry , phase (matter) , chemistry , high performance liquid chromatography , biochemistry , organic chemistry
Abstract 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 C x , apolipoprotein C‐I, apolipoprotein C‐III 2 , apolipoprotein C‐III 1 , apolipoprotein C‐III 0 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 and chromatography of pure protein standards.