Journal of Lipid Research, Vol. 11, 42-47, January 1970
Copyright © 1970 by Lipid Research, Inc.
Gradient-thickness thin-layer chromatography for the isolation and analysis of trace amounts of free fatty acids in large lipid samples
Nicolas G. Bazán Jr. and Cliffe D. Joel
Departments of Biological Chemistry and Psychiatry, Harvard Medical School, and Massachusetts Mental Health Center, Boston, Massachusetts 02115
A thin-layer chromatographic method for quantitative isolation of free fatty acids is described. This method appears to be more satisfactory than existing methods in offering the combination of advantages of specificity, simplicity, rapidity, reproducibility, accuracy, high sensitivity, and applicability as a preparative technique.
The method involves chromatography on a thin-layer plate on which the layer of Silica Gel G decreases linearly in thickness from 1000 µ at the base to 125 µ at the upper end. This gradient-thickness design allows the separation and densitometric quantitation of very small traces of free fatty acids from relatively large and complex lipid samples in a single chromatographic step.
The method has been shown to be applicable directly to the crude total lipid extracts of several mammalian tissues. It appears to generate little if any artifactual free fatty acids from the breakdown of complex lipids, in contrast to the undesirable behavior of silicic acid columns in this respect.
Gradient-thickness thin-layer chromatography promises to be useful for the quantitative isolation of trace amounts not only of other types of lipids but also of classes of compounds other than lipids.
Supplementary key words charring • quantitative densitometry • preparative chromatography • protection against peroxidation • artifactual free fatty acids • brain • liver • heart • blood plasma • coenzyme Q • triglycerides • hydrocarbons
Submitted on June 19, 1968
Revised on August 7, 1969
Accepted on September 15, 1969
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