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A method for separation of phosphatidylcholine, triacylglycerol, non-esterified fatty acids and cholesterol esters from plasma by solid-phase extraction*

Published online by Cambridge University Press:  09 March 2007

Graham C. Burdge ,
Paul Wright ,
Amanda E. Jones  and
Stephen A. Wootton
Graham C. Burdge*
Affiliation:
Institute of Human Nutrition, University of Southampton, Southampton, UK
Paul Wright
Affiliation:
Institute of Human Nutrition, University of Southampton, Southampton, UK
Amanda E. Jones
Affiliation:
Institute of Human Nutrition, University of Southampton, Southampton, UK
Stephen A. Wootton
Affiliation:
Institute of Human Nutrition, University of Southampton, Southampton, UK
*
†Corresponding author: Dr Graham C. Burdge, fax +44 2380 794945, email g.c.burdge@soton.ac.uk
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Abstract

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Efficient isolation of individual lipid classes is a critical step in the analysis of plasma and lipoprotein fatty acid compositions. Whilst good separations of total lipid extracts are possible by TLC, this method is time consuming and a major rate-limiting step when processing large numbers of specimens. A method for rapid separation of phosphatidylcholine (PC), non-esterified fatty acids (NEFA), cholesterol ester (CE) and triacylglycerol (TAG) from total plasma lipid extracts by solid-phase extraction (SPE) using aminopropyl silica columns has been developed and validated. Following initial separation of polar and neutral lipids, individual classes were isolated by application of solvents with increasing polarity. Recoveries for combined plasma extraction with chloroform–methanol and SPE were (%): PC 74·2 (SD 7·5), NEFA 73·6 (sd 8·3), CE 84·9 (sd 4·9), and TAG 86·8 (sd 4·9), which were significantly greater for TAG and NEFA than by TLC (P<0·001). Both GC–flame ionisation detector and GC-MS analysis of fatty acid methyl esters demonstrated that there was no cross-contamination between lipid classes. Measurements of repeatability of fatty acid composition for TAG, PC, CE and NEFA fractions showed similar CV for each fatty acid. The magnitude of the CV appeared to be related inversely to the fractional fatty acid concentration, and was greatest at concentrations of less than 1 g/100 g total fatty acids. There was no evidence of selective elution of individual fatty acid or CE species. In conclusion, this method represents an efficient, rapid alternative to TLC for isolation of these lipid classes from plasma.

Keywords

LipidsSolid-phase extractionAnalytical methodology
Type
Research Article
Information
British Journal of Nutrition , Volume 84 , Issue 5 , November 2000 , pp. 781 - 787
Copyright
Copyright © The Nutrition Society 2000

Footnotes

*

These data have been described previously, in part, in an abstract (Wright P, Burdge GC, Jones AE & Wooton SA (2000) Purification of the major plasma lipid classes by solid-phase extraction on aminopropyl silica cartridges. Proceedings of the Nutrition Society 52, 21A).

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