Separation of neutral lipid, free fatty acid and phospholipid classes by normal phase HPLC.

Abstract:

:Normal phase high performance liquid chromatography methods are described for the separation of neutral lipid, fatty acid and five phospholipid classes using spectrophotometric detection at 206 nm. Separations were accomplished in less than 10 min for each lipid class. A mobile phase consisting of hexane/methyltertiarybutylether/acetic acid (100:5:0.02) proved effective in separating cholesteryl ester and triglyceride with recoveries of 100% for radiolabeled cholesteryl oleate and 98% for radiolabeled triolein. Free fatty acid and cholesterol were separated by two different mobile phases. The first, hexane/methyltertiarybutylether/acetic acid (70:30:0.02) effectively separated free fatty acids and cholesterol, but did not separate cholesterol from 1,2-diglyceride. A mobile phase consisting of hexane/isopropanol/acetic acid (100:2:0.02) effectively separated free fatty acid, cholesterol, 1,2-diglyceride and 1,3-diglyceride. Recoveries of oleic acid and cholesterol were 100% and 97%, respectively. Five phospholipid classes were separated using methyltertiarybutylether/methanol/aqueous ammonium acetate (pH 8.6) (5:8:2) as the mobile phase. The recoveries of phosphatidylinositol, phosphatidylethanolamine, phosphatidylcholine, sphingomyelin and lysophosphatidylcholine were each greater than 96%.

journal_name

Lipids

journal_title

Lipids

authors

Hamilton JG,Comai K

doi

10.1007/BF02535282

subject

Has Abstract

pub_date

1988-12-01 00:00:00

pages

1150-3

issue

12

eissn

0024-4201

issn

1558-9307

journal_volume

23

pub_type

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