Profiling of non-esterified fatty acids in human plasma using liquid chromatography-electron ionization mass spectrometry |
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Authors: | Helga Trufelli Giorgio Famiglini Veronica Termopoli Achille Cappiello |
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Institution: | 1.Dipartimento di Scienze della Terra, della Vita e dell’Ambiente,Università di Urbino,Urbino,Italy |
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Abstract: | This paper focuses on the development of a novel approach to analyze underivatized fatty acids in human plasma. The method
is based on liquid–liquid extraction followed by reversed phase liquid chromatography coupled to direct-electron ionization
mass spectrometry (LC-Direct-EI-MS). The assay is validated. Calibrations show satisfactory linearity and precision in the
investigated range of linearity. Recoveries span from 75% to 104%. The method limits of detection, varying from 0.53 to 5.35 μM,
are satisfactory for the quantitation of non-esterified fatty acids (NEFAs) in plasma at physiological levels. The method
has been successfully applied to the NEFAs profiling of plasma samples from healthy adult volunteers and subjects affected
by diabetes mellitus. Compared with published protocols based on gas chromatography–mass spectrometry and liquid chromatography
coupled to electrospray ionization mass spectrometry, this method does not require derivatization and does not show matrix
effects, thus simplifying sample preparation procedure and reducing the total time of analysis to approximately 90 min. In
addition, Direct-EI-MS allows the acquisition of high-quality NIST library-matchable EI spectra, allowing an easy-to-obtain
identification of the target NEFAs. |
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