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Free fatty acid determination in plasma by GC-MS after conversion to Weinreb amides

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Abstract

Circulating free fatty acids (FFAs) play important physiological roles as contributing components in cellular structure as well as energy utilization. Elevated levels of circulating FFAs are associated with metabolic aberrations in humans. FFAs differ in chain length and saturation and may be altered in metabolically dysregulated conditions, such as type 2 diabetes mellitus. Potentially, alterations in circulating levels of specific FFAs could also be important in terms of prognostic value. Various methods have been used to analyze FFAs. In this study, a straightforward and accurate method for the determination of FFAs in plasma has been established and evaluated, through conversion of plasma FFAs into acid fluorides followed by conversion to Weinreb amides (dimethylamide). The method is mild, efficient, selective, and quantitative for FFAs, when analyzed with capillary gas chromatography tandem mass spectrometry. Standard curves were linear over the range of 1,000–20,000 ng/mL with a correlation coefficient (r 2) of 0.998, and coefficient of variation of triplicate analysis was <10 %. The gas chromatography–mass spectrometry (GC-MS) technique was reproducible and repeatable, and recoveries were above 90 %. From the generated MS spectra, five specific FFAs were identified. An explicit interest was the quantification of palmitate (C16:0) and palmitoleate (C16:1), which have been connected with detrimental and positive effects on the insulin-producing beta cells, respectively. The results demonstrate the suitability of Weinreb amides for efficient and rapid isolation of FFAs in plasma, prior to quantitative GC-MS analysis. We suggest that the method can be used as a routine standardized way of quantifying FFAs.

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Acknowledgments

We would like to acknowledge Professor Paresh Dutta (Department of Food Science, Swedish Agricultural University, Uppsala, Sweden) for his help with the instrumentation. The Swedish Research Council (grants 621-2008-3562, 621-2011-4423 (J.B.) and 72X-14019 (P.B.)), EC FP7-project Beta-JUDO (grant 279153; P.B., J.B., and A.F.), and the Uppsala Regional Research Council (A.F. and P.B.) are gratefully acknowledged for financial support.

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Authors and Affiliations

  1. Analytical Chemistry, Department of Chemistry—Biomedical Center and Science for Life Laboratory, Uppsala University, Box 599, 751 24, Uppsala, Sweden

    Sarojini J. K. A. Ubhayasekera & Jonas Bergquist

  2. Department of Medical Cell Biology, Uppsala University, Box 571, 751 23, Uppsala, Sweden

    Johan Staaf & Peter Bergsten

  3. Department of Women’s and Children’s Health, Uppsala University, 751 85, Uppsala, Sweden

    Johan Staaf & Anders Forslund

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  1. Sarojini J. K. A. Ubhayasekera
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  2. Johan Staaf
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  3. Anders Forslund
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  4. Peter Bergsten
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  5. Jonas Bergquist
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Corresponding author

Correspondence to Sarojini J. K. A. Ubhayasekera.

Additional information

Awarded an ABC Poster Prize on the occasion of the “21st Analysdagarna” held in Uppsala, Sweden from June 11–13, 2012.

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Ubhayasekera, S.J.K.A., Staaf, J., Forslund, A. et al. Free fatty acid determination in plasma by GC-MS after conversion to Weinreb amides. Anal Bioanal Chem 405, 1929–1935 (2013). https://doi.org/10.1007/s00216-012-6658-3

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  • DOI: https://doi.org/10.1007/s00216-012-6658-3

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