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Direct Microwave Transesterification of Fingertip Prick Blood Samples for Fatty Acid Determinations

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Lipids

Abstract

Omega-3 polyunsaturated fatty acid (PUFA) dietary intakes and tissue levels are positively associated with various health benefits. The development of cost efficient, high throughput methodologies would enable research in large clinical and population studies, and clinical fatty acid profiling. Microwave heating for the transesterification of blood fatty acids was examined. Samples were collected by venous puncture and fingertip prick onto chromatography paper. Aliquots of serum, plasma, erythrocytes and whole blood were prepared from venous blood. Boron trifluoride in methanol was used for transesterification but sample preparation and heating varied. Fatty acid determinations and markers of omega-3 fatty acid status including the sum of eicosapentaenoic acid and docosahexaenoic acid, the ratio of total n-3 PUFA to n-6 PUFA, and the percentage of n-3 highly unsaturated fatty acids (HUFA, ≥20 carbons and ≥3 carbon–carbon double bonds) in total HUFA were compared. Quantitative determinations indicate that microwave transesterification results in significantly lower estimates of monounsaturates and polyunsaturates, possibly through incomplete transesterification of triacylglycerols. However, qualitative estimates of omega-3 fatty acid status were relatively similar. Fingertip prick blood collection combined with direct transesterification by microwave may be a very rapid method to estimate omega-3 fatty acid status for selected applications.

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Abbreviations

BF3/MeOH:

Boron trifluoride in methanol

BHT:

Butylated hydroxytoluene

DHA:

Docosahexaenoic acid (22:6n-3)

DMA:

Dimethyl acetal

EDTA:

Ethylene diaminetetraacetic acid

EPA:

Eicosapentaenoic acid (20:5n-3)

FAME:

Fatty acid methyl ester

HUFA:

Highly unsaturated fatty acids

PUFA:

Polyunsaturated fatty acid

TLC:

Thin layer chromatography

17:0 PC:

1,2-Diheptadecanoyl-sn–glycero-3-phosphocholine

17:0 TG:

Triheptadecanoin

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Acknowledgements

Operating funds were provided by a start-up award from the University of Waterloo and infrastructure was purchased through a Canada Foundation of Innovation and the Ontario Research Fund matching grants. K.D.S. received partial salary support through a GENESIS Young Investigator Award and J.M.A held a NSERC Undergraduate Student Research Award for a portion of the study.

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Authors and Affiliations

  1. Department of Kinesiology, Laboratory of Nutritional and Nutraceutical Research, University of Waterloo, N2L 3G1, Waterloo, ON, Canada

    Jeff M. Armstrong, Adam H. Metherel & Ken D. Stark

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  1. Jeff M. Armstrong
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  2. Adam H. Metherel
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  3. Ken D. Stark
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Correspondence to Ken D. Stark.

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Armstrong, J.M., Metherel, A.H. & Stark, K.D. Direct Microwave Transesterification of Fingertip Prick Blood Samples for Fatty Acid Determinations. Lipids 43, 187–196 (2008). https://doi.org/10.1007/s11745-007-3141-6

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