DOI QR코드

DOI QR Code

Simultaneous Determination of Plasma Lactate, Pyruvate, and Ketone Bodies following tert-Butyldimethylsilyl Derivatization using GC-MS-SIM

  • Yoon, Hye-Ran (Biomedical & Pharmaceutical Analysis Lab, College of Pharmacy, Duksung Women's University)
  • 투고 : 2015.12.15
  • 심사 : 2015.12.28
  • 발행 : 2015.12.31

초록

Lactate and ketone bodies are considered biological markers for ketosis and several inherited metabolic disorders. In the current study, the specific ratios of lactate and ketone bodies as analytical tools for differential diagnosis of various lactic acidosis were devised. The study included a protein precipitation step following tert-butyldimethylsilyl derivatisation. Total run time was approximately 30 min including sample preparation and GS/MS analysis. The limits of detection were below 0.1 pg/mL over the targeted 4 analytes. The calibration curve was linear over the concentration range of $0.001{\sim}250{\mu}g/mL$ for pyruvate, beta-hydroxybutyrate, and acetoacetate ($R^2$ > 0.99). Inter-day accuracy and precision were 87.7~94.8% with RSD of 2.5~5.7% at 2 levels. Absolute recoveries (%) of target analytes were 87.0~98.4%. The method was validated for the quantification of lactate and ketone bodies for differentiation of lactic acidosis.

키워드

참고문헌

  1. Amathieu R, Triba MN, Nahon P, Bouchemal N, Kamoun W, Haouache H, Trinchet JC, Savarin P, Le Moyec L, Dhonneur G. Serum $^1H$-NMR metabolomic fingerprints of acute-on-chronic liver failure in intensive care unit patients with alcoholic cirrhosis. PLoS One. 2014. 19: e89230.
  2. Bhattacharya M, Fuhrman L, Ingram A, Nickerson KW, Conway T. Single-run separation and detection of multiple metabolic intermediates by anion-exchange high-performance liquid chromatography and application to cell pool extracts prepared from Escherichia coli. Anal Biochem. 1995. 232: 98-106. https://doi.org/10.1006/abio.1995.9954
  3. Chaves Das Heves, Vasconceios AM. Capillary gas chromatography of amino acids, including asparagine and glutamine: sensitive gas chromatographic-mass spectrometric and selected ion monitoring gas chromatographic-mass spectrometric detection of the N,O(S)-tert-butyldimethylsilyl derivatives. J Chromatogr. 1987. 392: 249-258. https://doi.org/10.1016/S0021-9673(01)94270-0
  4. Constantin-Teodosiu D, Cederblad G, Hultman E. A sensitive radioisotopic assay of pyruvate dehydrogenase complex in human muscle tissue. Anal Biochem. 1991. 198: 347-351. https://doi.org/10.1016/0003-2697(91)90437-X
  5. Dahl SR, Olsen KM, Strand DH. Determination of gamma-hydroxy butyrate (GHB), beta-hydroxybutyrate (BHB), pregabalin, 1,4-butane-diol (1,4BD) and gamma-butyrolactone (GBL) in whole blood and urine samples by UPLC-MSMS. J Chromatogr B. 2012. 885: 37-42.
  6. Elliott S, Smith C, Cassidy D. The post-mortem relationship betwee betahydroxybutyrate (BHB), acetone and ethanol in ketoacidosis. Forensic Sci Int. 2010. 198: 53-57. https://doi.org/10.1016/j.forsciint.2009.10.019
  7. Felby S, Nielsen E. Determination of ketone bodies in post-mortem blood by headspace gas chromatography. Forensic Sci Int. 1994. 64: 83-88. https://doi.org/10.1016/0379-0738(94)90216-X
  8. Hassan HMA, Cooper GAA. Determination of $\beta$-hydroxybutyrate in blood and urine using gas chromatography-mass spectrometry. J Anal Toxicol. 2009. 33: 502-507. https://doi.org/10.1093/jat/33.8.502
  9. Hoffmann G, Aramak S, BIum-Hoffmann E, Nyhan WL, Sweetman L. Quantitative analysis for organic acids in biological samples: batch isolation followed by gas chromatographic-mass spectrometric analysis. Clin Chem. 1989. 35: 587-595.
  10. Holm KMD, Linnet K, Rasmussen BS, Pedersen AJ. Determination of ketone bodies in blood by headspace gas chromatography-mass spectrometry. J Anal Toxicol. 2010. 34: 549-554. https://doi.org/10.1093/jat/34.9.549
  11. Hur H, Paik MJ, Xuan Y, Nguyen DT, Ham IH, Yun J, Cho YK, Lee G, Han SU. Quantitative measurement of organic acids in tissues from gastric cancer patients indicates increased glucose metabolism in gastric cancer. PLoS One. 2014. 9: e98581. https://doi.org/10.1371/journal.pone.0098581
  12. Kapoor P, Mandal B, Chowdhury U, Singh S, Kiran U. Changes in myocardial lactate, pyruvate and lactate-pyruvate ratio during cardiopulmonary bypass for elective adult cardiac surgery: Early indicator of morbidity. J Anaesthesiol Clin Pharmacol. 2011. 27: 225-232. https://doi.org/10.4103/0970-9185.81819
  13. Karlsson J, Jacobs I, Sjödin B, Tesch P, Kaiser P, Sahl O, Karlberg B. Semi-automatic blood lactate assay: experiences from an exercise laboratory. Int J Sports Med. 1983. 4: 52-55. https://doi.org/10.1055/s-2008-1026016
  14. Kimura M, Kobayashi K, Matsuoka A, Hayashi K. Head-space gas-chromatographic determination of 3-hydroxybutyrate in plasma after enzymatic reactions, and the relationship among the three ketone bodies. Clin Chem. 1985. 31: 596-598.
  15. Lee K, Berthiaume F, Stephanopoulos GN, Yarmush ML. Metabolic flux analysis: a powerful tool for monitoring tissue function. Tissue Eng. 1999. 5: 347-368. https://doi.org/10.1089/ten.1999.5.347
  16. Lennart Lundholm, Ella Mohme-Lundholm and Nandor Vamos Lactic Acid Assay with L(+)lactic Acid Dehydrogenase from Rabbit Muscle. Acta Physiol Scand. 1963. 58: 243-249. https://doi.org/10.1111/j.1748-1716.1963.tb02645.x
  17. Levy B, Sadoune LO, Gelot AM, Bollaert P, Nabet PG, Larcan A. Evolution of lactate/pyruvate and arterial ketone body ratios in the early course of catecholamine-treated septic shock. Crit Care Med. 2000. 28: 114-119. https://doi.org/10.1097/00003246-200001000-00019
  18. Lloyd B, Burrin J, Smythe P, Alberti KG. Enzymic fluorometric continuous-flow assays for blood glucose, lactate, pyruvate, alanine, glycerol, and 3-hydroxybutyrate. Clin Chem. 1978. 24: 1724-1729.
  19. Lorenz, I. Hartmann, A. Gentile Determination of D-lactate in calf serum samples - an automated enzymatic assay. Comp Clin Path. 2003. 12: 169-171. https://doi.org/10.1007/s00580-003-0499-6
  20. Nissen S, Van Koevering M, Webb D. Analysis of ${\beta}$-hydroxy- ${\beta}$-methyl butyrate in plasma by gas chromatography and mass spectrometry. Anal Biochem. 1990. 188: 17-19. https://doi.org/10.1016/0003-2697(90)90522-B
  21. Shimizu I, Ishikawa E, Fukuda T, Okazaki S. Blood glucose, urinary glucose, blood ketone bodies, urine ketone bodies, blood lactic acid, and blood pyruvic acid. Nihon Rinsho. 2012. 70: 433-437.
  22. Shinde S, Golam K, Patil ND, Kumar P. Perioperative blood lactate levels, pyruvate levels and lactate-pyruvate ratio in children undergoing cardiopulmonary bypass for congenital heart disease. Indian J Crit Med. 2005. 9: 145-150. https://doi.org/10.4103/0972-5229.19680
  23. Slamet T, Lim PKC, Mak JW, Storey J, Noor RA. Sensitivity of Malaysian isolates of Plasmodium falciparum to a number of antimalarials using the WHO in vitro micro test. Tropical Biomedicine. 1991. 8: 85-87.
  24. Vassault A, Bonnefont JP, Specola N, Saudubray JM. Lactate, pyruvate, and ketone bodies. In Techniques in Diagnostic Human Biochemical Genetics - a Laboratory Manual, Hommes FA (ed.). New York: Wiley-Liss. 1991: 28-86.
  25. Villas-Boas SG, Mas S, Akesson M, Smedsgaard J, Nielsen J. Mass spectrometry in metabolome analysis. Mass Spectrom Rev. 2005. 24: 613-646. https://doi.org/10.1002/mas.20032
  26. Vogel C, Wessel E, Siesler H. W. FT-IR imaging spectroscopy of phase separation in blends of poly(3-hydroxybutyrate) with poly(L-lactic acid) and poly(epsilon-caprolactone). Biomacromolecules. 2008. 9: 523-527. https://doi.org/10.1021/bm701035p
  27. Yamato S, Shinohara K, Nakagawa S, Kubota A, Inamura K, Watanabe G, Hirayma S, Miida T, Ohtas. High performance liquid chromatography determination of ketone bodies in human plasma by derivatization with p-nitrobenzene diazonium fluoroborate. Anal Biochem 2009. 384: 145-150. https://doi.org/10.1016/j.ab.2008.09.017
  28. Yarmush ML,Banta S. Metabolic engineering: Advancesinmodeling and intervention in health and disease. Annu Rev Biomed Eng. 2003. 5: 349-381. https://doi.org/10.1146/annurev.bioeng.5.031003.163247
  29. Zhua A, Romerob R, Howard R. Pettya, A sensitive fluorimetric assay for pyruvate Anal Biochem. 2010. 396: 146-151. https://doi.org/10.1016/j.ab.2009.09.017