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

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Abstract

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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References

  1. Brustugun J, Tonnesen HH, Edge R, Navaratnam S (2005) Formation and reactivity of free radicals in 5-hydroxymethyl-2-furaldehyde - the effect on isoprenaline photostability. J Photochem Photobiol B 79:109–119

    Article  CAS  Google Scholar 

  2. Karlsson B (1962) Kurzes Lehrbuch der Biochemie für Mediziner und Naturwissenschaftler. Thieme, Stuttgart

    Google Scholar 

  3. Kronberger L, FinK E (1968) Pyruvic acid, lactic acid and alpha-ketoglutaric acid in carcinoma patients. Med Klin 63:944–948

    CAS  Google Scholar 

  4. Schaur RJ, Schreibmayer W, Semmelrock HJ, Tillian HM, Schauenstein E (1979) Tumor host relations. II. Increase of alpha-ketoglutarate in whole blood and urine and hypoalbuminemia of rats bearing the solid rhabdomyosarcoma BA 1112 and the ascitic or solid forms of Walker-carcinoma 256 and Yoshida-sarcoma. J Cancer Res Clin Oncol 93:293–300

    Article  CAS  Google Scholar 

  5. Kronberger L, Semmelrock HJ, Schaur RJ, Schauenstein E, Schreibmayer W, Fink E (1980) Tumor host relations. VI. Is alpha-ketoglutarate a tumor marker? Association with tumor extent in humans–correlation with tumor size in rats. J Cancer Res Clin Oncol 97:295–299

    Article  CAS  Google Scholar 

  6. Michail K, Juan H, Maier A, Matzi V, Greilberger J, Wintersteiger R (2007) Development and validation of a liquid chromatographic method for the determination of hydroxymethylfurfural and alpha-ketoglutaric acid in human plasma. Anal Chim Acta 581:287–297

    Article  CAS  Google Scholar 

  7. Matzi V, Lindenmann J, Muench A, Greilberger J, Juan H, Wintersteiger R et al (2007) The impact of preoperative micronutrient supplementation in lung surgery. A prospective randomized trial of oral supplementation of combined alpha-ketoglutaric acid and 5-hydroxymethylfurfural. Eur J Cardiothorac Surg 32:776–782

    Article  Google Scholar 

  8. Kong Z, Shinohara L, Mitsuiki M, Murakami H, Omura H (1989) Desmutagenicity of furan compounds towards some mutagens. Agricult Bioll Chem 53:2073–2079

    CAS  Google Scholar 

  9. Abdulmalik O, Safo MK, Chen Q, Yang J, Brugnara C, Ohene-Frempong K et al (2005) 5-hydroxymethyl-2-furfural modifies intracellular sickle haemoglobin and inhibits sickling of red blood cells. Br J Haematol 128:552–561

    Article  CAS  Google Scholar 

  10. Teixidó E, Santos FJ, Puignou L, Galceran MT (2006) Analysis of 5-hydroxymethylfurfural in foods by gas chromatography–mass spectrometry. J Chromatogr A 1135:85–90

    Article  Google Scholar 

  11. Rupp HS, Turnipseed SB (2000) Confirmation of patulin and 5-hydroxymethylfurfural in apple juice by gas chromatography/mass spectrometry. J AOAC Int 83:612–620

    CAS  Google Scholar 

  12. Laplante A, Comte B, Des Rosiers C (1995) Assay of blood and tissue oxaloacetate and alphaketoglutarate by isotope dilution gas chromatography–mass spectrometry. Anal Biochem 224:580–587

    Article  CAS  Google Scholar 

  13. Rocchiccio F, Leroux JP, Cartier P (1981) Quantitative gas chromatography–chemical ionization mass spectrometry of 2-ketoglutarate from urine as its O-trimethylsilylquinoxalinol derivative. J Chromatogr 226:325–332

    Article  Google Scholar 

  14. Lo Coco F, Novelli V, Valentini V, Ceccon L (1997) High-performance liquid chromatographic determination of 2-furaldehyde and 5-hydroxymethyl-2-furaldehyde in fruit juices. J Chromatogr Sci 35:578–583

    CAS  Google Scholar 

  15. Traore F, Pianetti GA, Dallery L, Tod M, Chalom J, Farinotti R et al (1993) Determination of picomole amounts of carbonyls as luminarin hydrazones by high-performance liquid chromatography with fluorescence detection. Chromatographia 36:96–104

    Article  CAS  Google Scholar 

  16. Lange M, Mályusz M (1994) Fast method for the simultaneous determination of 2-oxo acids in biological fluids by high-performance liquid chromatography. J Chromatogr B Biomed Appl 662:97–102

    Article  CAS  Google Scholar 

  17. Nohta H, Sonoda J, Yoshida H, Satozono H, Ishida J, Yamaguchi M (2003) Liquid chromatographic determination of dicarboxylic acids based on intramolecular excimer-forming fluorescence derivatization. J Chromatogr A 1010:37–44

    Article  CAS  Google Scholar 

  18. Ciba-Geigy. Wissenschaftliche Tabellen Geigy. Teilband Hämatologie und Humangenetik. Ciba-Geigy, Basel; 1979.

  19. Kristensen NB, Jungvid H, Fernandez JA, Pierzynowski SG (2002) Absorption and metabolism of α-ketoglutarate in growing pigs. J Anim Physiol Anim Nutr 86:239–245

    Article  CAS  Google Scholar 

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Authors and Affiliations

  1. Research Unit of Osteology and Analytical Mass Spectrometry, University Children’s Hospital, Medical University Graz, Auenbruggerplatz 30, 8036, Graz, Austria

    Bernhard M. Wagner, Werner Windischhofer & Hans J. Leis

  2. Institute of Pharmaceutical Sciences, Karl-Franzens-University Graz, Schubertstraße 1, 8010, Graz, Austria

    Fabrizio Donnarumma & Reinhold Wintersteiger

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  1. Bernhard M. Wagner
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  2. Fabrizio Donnarumma
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  3. Reinhold Wintersteiger
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  4. Werner Windischhofer
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  5. Hans J. Leis
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Correspondence to Hans J. Leis.

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Wagner, B.M., Donnarumma, F., Wintersteiger, R. et al. Simultaneous quantitative determination of α-ketoglutaric acid and 5-hydroxymethylfurfural in human plasma by gas chromatography–mass spectrometry. Anal Bioanal Chem 396, 2629–2637 (2010). https://doi.org/10.1007/s00216-010-3479-0

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  • DOI: https://doi.org/10.1007/s00216-010-3479-0

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