Hostname: page-component-7dd5485656-gs9qr Total loading time: 0 Render date: 2025-10-31T15:19:13.442Z Has data issue: false hasContentIssue false
  • English
  • Français

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
Rights & Permissions [Opens in a new window]

Abstract

Core share and HTML view are not available for this content. However, as you have access to this content, a full PDF is available via the ‘Save PDF’ action button.

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

Information

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).

References

Agren, JJ, Julkunen, A and Penttila, I (1992) Rapid separation of serum lipids for fatty acid analysis by a single aminopropyl column. Journal of Lipid Research 33, pp. 1871–1876.CrossRefGoogle ScholarPubMed
Bernhardt, TG, Cannistraro, PA, Bird, DA, Doyle, KM and Laposata, M (1996) Purification of fatty acid ethyl esters by solid-phase extraction and high performance liquid chromatography. Journal of Chromatography B 675, pp. 189–196.CrossRefGoogle ScholarPubMed
Caesar, PA, Wilson, SJ, Normand, ICS and Postle, AD (1988) A comparison of the specificity of phosphatidylcholine synthesis by human fetal lung maintained in either organ or organotypic culture. Biochemical Journal 253, pp. 451–457.CrossRefGoogle ScholarPubMed
Folch, J, Lees, M and Sloane-Stanley, GH (1957) A simple method for the isolation and purification of total lipides from animal tissues. Journal of Biological Chemistry 226, pp. 497–509.CrossRefGoogle ScholarPubMed
Fosbrooke, AS and Tamir, I (1968) A modified method for the preparation of methyl esters of a mixture of medium-chain and long-chain fatty acids. Application to the determination of serum triglyceride and non-esterified fatty acid composition and concentration by gas-liquid chromatography. Clinica Chimica Acta 20, pp. 517–522.CrossRefGoogle Scholar
Houwelingen, ACv, Foreman-van, Drongelen, MMHP, Nicolini, U, Nicolaides, KH, Al, MDM, Kester, ADM and Hornstra, G (1996) Essential fatty acid status of fetal plasma phospholipids: similar to postnatal values obtained at comparable gestational ages. Early Human Development 46, pp. 141–152.CrossRefGoogle ScholarPubMed
Hoving, EB, Jansen, G, Volmer, M, Van Doormaal, JJ and Muskiet, FAJ (1998) Profiling of plasma cholesterol ester and triglyceride fatty acids as their methyl esters by capillary gas chromatography, preceded by a rapid aminopropyl-silica column chromatographic separation of lipid classes. Journal of Chromatography 434, pp. 395–409.CrossRefGoogle Scholar
Jensen, RG, Clark, RM, Gerrior, SA, Fey, MB and Gotto, AM (1979) Cholesterol ester and triacylglycerol fatty acids in type V hyperlipidaemia. Lipids 14, pp. 691–694.CrossRefGoogle Scholar
Kim, HY and Salem, N (1990) Separation of lipid classes by solid phase extraction. Journal of Lipid Research 31, pp. 2285–2289.CrossRefGoogle ScholarPubMed