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Improved Extraction of Saturated Fatty Acids but not Omega-3 Fatty Acids from Sheep Red Blood Cells Using a One-Step Extraction Procedure

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Lipids

Abstract

Several methods are available to extract total lipid and methylate fatty acids from a range of samples including red blood cells (RBC). Fatty acid analysis of human RBC can be undertaken using a two-step extraction and methylation or a combined one-step extraction and methylation procedure. The lipid composition of sheep RBC differs significantly from that of humans and may affect their extraction. The purpose of the current study was to examine the efficiency of extraction of lipid and detection of fatty acids from sheep RBC using a one-step procedure. Fatty acids were analysed using a one-step extraction and methylation procedure using methanol:toluene and acetyl chloride in comparison with a two-step procedure involving extraction of lipid using chloroform:methanol and separate methylation. Concentrations of saturated fatty acids including C16:0 and C18:0 were significantly higher (42.6 and 33.9 % respectively) following extraction using the one-step procedure compared with the two-step procedure. However, concentrations of some polyunsaturated fatty acids, including C20:5n-3 and C22:6n-3 were not significantly different between either procedure. The improved detection of fatty acids may be related to the ability of different solvents to extract different lipid fractions. The differential extraction of lipids and detection of fatty acids from sheep RBC may have important implications in studies examining the effect of dietary treatment on the possible health benefits of fatty acids.

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Abbreviations

FAME:

Fatty acid methyl esters

FID:

Flame ionisation detector

LoD:

Limit of detection

LCn-3PUFA:

Long-chain omega-3 polyunsaturated fatty acids

LCn-6PUFA:

Long-chain omega-6 polyunsaturated fatty acids

MUFA:

Monounsaturated fatty acids

RBC:

Red blood cells

SFA:

Saturated fatty acids

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Acknowledgments

We thank Greg Clark, Steven Huckell, Michael Loiterton, Rex Edis, Craig Rodham, John Wilkins, Susan Robertson, the Gulliver family, Jessica Rose, John Broster and Dr Stephanie Knott for technical assistance during the conduct of the study. We also thank Jamie Ayton for providing assistance with the gas chromatography. This study was conducted without dedicated external funding. This study was supported by in-kind provision of equipment by the Diagnostic and Analytical Systems division of NSW DPI.

Conflict of interest

None.

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Authors and Affiliations

  1. NSW Department of Primary Industries, Wagga Wagga Agricultural Institute, Pine Gully Rd, Wagga Wagga, NSW, 2650, Australia

    Edward H. Clayton, John W. Piltz & Richard G. Meyer

  2. EH Graham Centre for Agricultural Innovation, an alliance between NSW DPI and Charles Sturt University, Wagga Wagga, NSW, Australia

    Edward H. Clayton, Catherine E. Gulliver & John W. Piltz

  3. School of Animal and Veterinary Science, Charles Sturt University, Wagga Wagga, NSW, 2678, Australia

    Catherine E. Gulliver

  4. Future Farm Industries CRC, The University of Western Australia, Crawley, WA, 6009, Australia

    Catherine E. Gulliver

  5. Australian Livestock Research, Charlestown Rd, Kotara South, NSW, 2289, Australia

    Robert D. Taylor

  6. Hunter New England Area Health Service, John Hunter Hospital, Lookout Rd, New Lambton, NSW, 2305, Australia

    Robert J. Blake

  7. School of Biomedical Sciences, University of Newcastle, Callaghan, NSW, 2308, Australia

    Robert J. Blake

Authors
  1. Edward H. Clayton
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  2. Catherine E. Gulliver
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  3. John W. Piltz
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  4. Robert D. Taylor
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  5. Robert J. Blake
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  6. Richard G. Meyer
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Correspondence to Edward H. Clayton.

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Clayton, E.H., Gulliver, C.E., Piltz, J.W. et al. Improved Extraction of Saturated Fatty Acids but not Omega-3 Fatty Acids from Sheep Red Blood Cells Using a One-Step Extraction Procedure. Lipids 47, 719–727 (2012). https://doi.org/10.1007/s11745-012-3674-1

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  • DOI: https://doi.org/10.1007/s11745-012-3674-1

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