Simultaneous quantitative determination of α-ketoglutaric acid and 5-hydroxymethylfurfural in human plasma by gas chromatography–mass spectrometry

Wagner, Bernhard M.; Donnarumma, Fabrizio; Wintersteiger, Reinhold; Windischhofer, Werner; Leis, Hans J.
April 2010
Analytical & Bioanalytical Chemistry;Apr2010, Vol. 396 Issue 7, p2629
Academic Journal
α-Ketoglutaric acid (α-KG) and 5-hydroxymethylfurfural (5-HMF) are currently under investigation as promising cancer cell damaging agents. A method for the simultaneous quantitative determination of α-KG and 5-HMF in human plasma was established for screening these compounds in human plasma. Plasma samples were directly treated with O-(2,3,4,5,6-pentafluorobenzyl) hydroxylamine hydrochloride to form the corresponding oximes, thus facilitating subsequent liquid–liquid extraction. After formation of the trimethylsilyl ethers, samples were analyzed by gas chromatography with electron ionization mass spectrometry. Stable isotope labeled standards were used, the preparation of 13C6-5-HMF is described. Limits of quantitation were set to 0.938 µg/mL for α-KG and 0.156 µg/mL for 5-HMF. Inter-day accuracy was ≤93.7% (α-KG) and ≤92.8% (5-HMF). Inter-day precision was ≤6.0% (α-KG) and ≤4.6% (5-HMF). The method has been successfully applied to pharmacokinetic profiling of the compounds after intravenous application.


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