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

A method for separation of phosphatidylcholine, triacylglycerol, non-esterified fatty acids and cholesterol esters from plasma by solid-phase extraction.
A method for separation of phosphatidylcholine, triacylglycerol, non-esterified fatty acids and cholesterol esters from plasma by solid-phase extraction.
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 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.
0007-1145
781-787
Burdge, G.C.
09d60a07-8ca1-4351-9bf1-de6ffcfb2159
Wright, P.
56f297c9-9693-463d-ad60-f23cddc80250
Jones, A.E.
61f65ad1-ae5c-47f6-b159-63aa6a5178b5
Wootton, S.A.
bf47ef35-0b33-4edb-a2b0-ceda5c475c0c
Burdge, G.C.
09d60a07-8ca1-4351-9bf1-de6ffcfb2159
Wright, P.
56f297c9-9693-463d-ad60-f23cddc80250
Jones, A.E.
61f65ad1-ae5c-47f6-b159-63aa6a5178b5
Wootton, S.A.
bf47ef35-0b33-4edb-a2b0-ceda5c475c0c

Burdge, G.C., Wright, P., Jones, A.E. and Wootton, S.A. (2000) A method for separation of phosphatidylcholine, triacylglycerol, non-esterified fatty acids and cholesterol esters from plasma by solid-phase extraction. British Journal of Nutrition, 84 (5), 781-787. (doi:10.1017/S0007114500002154). (PMID:11177194)

Record type: Article

Abstract

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 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.

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Published date: November 2000
Organisations: Human Development & Health

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Local EPrints ID: 383839
URI: http://eprints.soton.ac.uk/id/eprint/383839
ISSN: 0007-1145
PURE UUID: d6f74cb4-230a-45bc-b574-9d6811bdee70
ORCID for G.C. Burdge: ORCID iD orcid.org/0000-0002-7665-2967

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Date deposited: 25 Nov 2015 14:16
Last modified: 07 Oct 2020 01:35

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Contributors

Author: G.C. Burdge ORCID iD
Author: P. Wright
Author: A.E. Jones
Author: S.A. Wootton

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