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Simple Methodology for the Quantitative Analysis of Fatty Acids in Human Red Blood Cells

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Abstract

In the last years, there has been an increasing interest in evaluating possible relations between fatty acid (FA) patterns and the risk for chronic diseases. Due to the long life span (120 days) of red blood cells (RBCs), their FA profile reflects a longer term dietary intake and was recently suggested to be used as an appropriate biomarker to investigate correlations between FA metabolism and diseases. Therefore, the aim of this work was to develop and validate a simple and fast methodology for the quantification of a broad range of FAs in RBCs using gas chromatography with flame ionization detector, as a more common and affordable equipment suitable for biomedical and nutritional studies including a large number of samples. For this purpose, different sample preparation protocols were tested and compared, including a classic two-step method (Folch method) with modifications and different one-step methods, in which lipid extraction and derivatization were performed simultaneously. For the one-step methods, different methylation periods and the inclusion of a saponification reaction were evaluated. Differences in absolute FA concentrations were observed among the tested methods, in particular for some metabolically relevant FAs such as trans elaidic acid and eicosapentaenoic acid. The one-step method with saponification and 60 min of methylation time was selected since it allowed the identification of a higher number of FAs, and was further submitted to in-house validation. The proposed methodology provides a simple, fast and accurate tool to quantitatively analyse FAs in human RBCs, useful for clinical and nutritional studies.

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Acknowledgments

This work received financial support from the European Union (FEDER funds through COMPETE) and National Funds (FCT, Fundação para a Ciência e Tecnologia) through project PTDC/SAU-ENB/116929/2010 and EXPL/EMS-SIS/2215/2013. ROR acknowledges PhD scholarship SFRH/BD/97658/2013 attributed by FCT (Fundação para a Ciência e Tecnologia).

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    Authors and Affiliations

    1. LCM-Laboratory of Catalysis and Materials-Associate Laboratory LSRE/LCM, Faculdade de Engenharia da Universidade do Porto (FEUP), R. Dr. Roberto Frias, 4200-465, Porto, Portugal

      Raquel O. Rodrigues

    2. Polytechnic Institute of Bragança, ESTiG/IPB, C. Sta. Apolónia, 5301-857, Bragança, Portugal

      Raquel O. Rodrigues, Helena Costa, Rui Lima & Joana S. Amaral

    3. CEFT, Faculdade de Engenharia da Universidade do Porto (FEUP), R. Dr. Roberto Frias, 4200-465, Porto, Portugal

      Rui Lima

    4. Mechanical Engineering Department, University of Minho, Campus de Azurém, 4800-058, Guimarães, Portugal

      Rui Lima

    5. REQUIMTE, Departamento de Ciências Químicas, Faculdade de Farmácia Universidade do Porto, Rua de Jorge Viterbo Ferreira no 228, 4050-313, Porto, Portugal

      Joana S. Amaral

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    1. Raquel O. Rodrigues
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    Correspondence to Joana S. Amaral.

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    R. O. Rodrigues and H. Costa have contributed equally to this work.

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    Rodrigues, R.O., Costa, H., Lima, R. et al. Simple Methodology for the Quantitative Analysis of Fatty Acids in Human Red Blood Cells. Chromatographia 78, 1271–1281 (2015). https://doi.org/10.1007/s10337-015-2947-2

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    • DOI: https://doi.org/10.1007/s10337-015-2947-2

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