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Journal of Lipid Research, Vol 38, 1482-1489, Copyright © 1997 by Lipid Research, Inc.
Quantification in the subnanomolar range of phospholipids and neutral lipids by monodimensional thin-layer chromatography and image analysis
JI Ruiz and B Ochoa
Department of Physiology, University of the Basque Country Medical School, Bilbao, Spain.
A sensitive, rapid, and simple method is presented for the simultaneous
quantitation of the major phospholipids and neutral lipids using an image
analyzer after separation by one-dimensional thin-layer chromatography. A
clear-cut separation of the lipids was achieved on one EDTA-impregnated
chromatoplate with five step-wise developments. An image was acquired, and
the integrated optical densities of the individual spots were quantitated
by a camera-equipped image analyzer against an internal standard of
cholesteryl formate. Calibration curves for each lipid followed linear or
hyperbolic functions permitting quantification of as little as 0.015 nmol
of lysophosphatidylcholine and as much as 7.50 nmol of triglyceride from a
single application. Quantitation of the major lipids in human plasma by TLC
and image analysis and standardized enzymatic methods were well correlated.
The method is suitable for routine analysis of biological samples having
lipid profiles as different as rat hepatocyte subcellular fractions, and
very low density lipoproteins secreted by rat hepatocyte suspensions.

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Copyright © 1997 by the American Society for Biochemistry and Molecular Biology.
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