Abstract
Background: The reactive α-oxoaldehydes glyoxal (GO), methylglyoxal (MGO) and 3-deoxyglucosone (3-DG) have been linked to diabetic complications and other age-related diseases. Numerous techniques have been described for the quantification of α-oxoaldehydes in blood or plasma, although with several shortcomings such as the need of large sample volume, elaborate extraction steps or long run-times during analysis. Therefore, we developed and evaluated an improved method including sample preparation, for the quantification of these α-oxoaldehydes in blood and plasma with ultra performance liquid chromatography tandem mass spectrometry (UPLC MS/MS).
Methods: EDTA plasma and whole blood samples were deproteinized using perchloric acid (PCA) and subsequently derivatized with o-phenylenediamine (oPD). GO, MGO and 3-DG concentrations were determined using stable isotope dilution UPLC MS/MS with a run-to-run time of 8 min. Stability of α-oxoaldehyde concentrations in plasma and whole blood during storage was tested. The concentration of GO, MGO and 3-DG was measured in EDTA plasma of non-diabetic controls and patients with type 2 diabetes (T2DM).
Results: Calibration curves of GO, MGO and 3-DG were linear throughout selected ranges. Recoveries of these α-oxoaldehydes were between 95% and 104%. Intra- and inter-assay CVs were between 2% and 14%.
Conclusions: To obtain stable and reliable α-oxoaldehyde concentrations, immediate centrifugation of blood after blood sampling is essential and the use of EDTA as anticoagulant is preferable. Moreover, immediate precipitation of plasma protein with PCA stabilized α-oxoaldehyde concentrations for at least 120 min. With the use of the developed method, we found increased plasma concentrations of GO, MGO and 3-DG in T2DM as compared with non-diabetic controls.
References
1. Han Y, Randell E, Vasdev S, Gill V, Gadag V, Newhook LA, et al. Plasma methylglyoxal and glyoxal are elevated and related to early membrane alteration in young, complication-free patients with type 1 diabetes. Mol Cell Biochem 2007;305:123–31.10.1007/s11010-007-9535-1Search in Google Scholar
2. Yamada H, Miyata S, Igaki N, Yatabe H, Miyauchi Y, Ohara T, et al. Increase in 3-deoxyglucosone levels in diabetic rat plasma. Specific in vivo determination of intermediate in advanced Maillard reaction. J Biol Chem 1994;269:20275–80.10.1016/S0021-9258(17)31987-7Search in Google Scholar
3. Thornalley PJ, Langborg A, Minhas HS. Formation of glyoxal, methylglyoxal and 3-deoxyglucosone in the glycation of proteins by glucose. Biochem J 1999;344:109–16.10.1042/bj3440109Search in Google Scholar
4. Baynes JW, Thorpe SR. Role of oxidative stress in diabetic complications: a new perspective on an old paradigm. Diabetes 1999;48:1–9.10.2337/diabetes.48.1.1Search in Google Scholar PubMed
5. Odani H, Shinzato T, Matsumoto Y, Usami J, Maeda K. Increase in three α,β-dicarbonyl compound levels in human uremic plasma: specific in vivo determination of intermediates in advanced Maillard reaction. Biochem Biophys Res Commun 1999;256:89–93.10.1006/bbrc.1999.0221Search in Google Scholar PubMed
6. Lu J, Randell E, Han Y, Adeli K, Krahn J, Meng QH. Increased plasma methylglyoxal level, inflammation, and vascular endothelial dysfunction in diabetic nephropathy. Clin Biochem 2011;44:307–11.10.1016/j.clinbiochem.2010.11.004Search in Google Scholar PubMed
7. Ogawa S, Nakayama K, Nakayama M, Mori T, Matsushima M, Okamura M, et al. Methylglyoxal is a predictor in type 2 diabetic patients of intima-media thickening and elevation of blood pressure. Hypertension 2010;56:471–6.10.1161/HYPERTENSIONAHA.110.156786Search in Google Scholar PubMed
8. Nakayama K, Nakayama M, Iwabuchi M, Terawaki H, Sato T, Kohno M, et al. Plasma α-oxoaldehyde levels in diabetic and nondiabetic chronic kidney disease patients. Am J Nephrol 2008;28:871–8.10.1159/000139653Search in Google Scholar PubMed
9. Nin JW, Jorsal A, Ferreira I, Schalkwijk CG, Prins MH, Parving H-H, et al. Higher plasma levels of advanced glycation end products are associated with incident cardiovascular disease and all-cause mortality in type 1 diabetes: a 12-year follow-up study. Diabetes Care 2011;34:442–7.10.2337/dc10-1087Search in Google Scholar PubMed PubMed Central
10. Srikanth V, Maczurek A, Phan T, Steele M, Westcott B, Juskiw D, et al. Advanced glycation end products and their receptor RAGE in Alzheimer’s disease. Neurobiol Aging 2011;32:763–77.10.1016/j.neurobiolaging.2009.04.016Search in Google Scholar PubMed
11. McLellan AC, Phillips SA, Thornalley PJ. The assay of methylglyoxal in biological systems by derivatization with 1,2-diamino-4,5-dimethoxybenzene. Anal Biochem 1992;206:17–23.10.1016/S0003-2697(05)80005-3Search in Google Scholar
12. Mittelmaier S, Funfrocken M, Fenn D, Fichert T, Pischetsrieder M. Identification and quantification of the glucose degradation product glucosone in peritoneal dialysis fluids by HPLC/DAD/MSMS. J Chromatogr B Analyt Technol Biomed Life Sci 2010;878:877–82.10.1016/j.jchromb.2010.02.004Search in Google Scholar PubMed
13. Nemet I, Varga-Defterdarovic L, Turk Z. Preparation and quantification of methylglyoxal in human plasma using reverse-phase high-performance liquid chromatography. Clin Biochem 2004;37:875–81.10.1016/j.clinbiochem.2004.05.024Search in Google Scholar PubMed
14. Neng NR, Cordeiro CA, Freire AP, Nogueira JM. Determination of glyoxal and methylglyoxal in environmental and biological matrices by stir bar sorptive extraction with in-situ derivatization. J Chromatogr A 2007;1169:47–52.10.1016/j.chroma.2007.08.060Search in Google Scholar PubMed
15. Dhar A, Desai K, Liu J, Wu L. Methylglyoxal, protein binding and biological samples: are we getting the true measure? J Chromatogr B Analyt Technol Biomed Life Sci 2009;877:1093–100.10.1016/j.jchromb.2009.02.055Search in Google Scholar PubMed
16. Mittelmaier S, Funfrocken M, Fenn D, Berlich R, Pischetsrieder M. Quantification of the six major α-dicarbonyl contaminants in peritoneal dialysis fluids by UHPLC/DAD/MSMS. Anal Bioanal Chem 2011;401:1183–93.10.1007/s00216-011-5195-9Search in Google Scholar PubMed
17. Randell EW, Vasdev S, Gill V. Measurement of methylglyoxal in rat tissues by electrospray ionization mass spectrometry and liquid chromatography. J Pharmacol Toxicol Methods 2005;51:153–7.10.1016/j.vascn.2004.08.005Search in Google Scholar PubMed
18. Kampf CJ, Bonn B, Hoffmann T. Development and validation of a selective HPLC-ESI-MS/MS method for the quantification of glyoxal and methylglyoxal in atmospheric aerosols (PM2.5). Anal Bioanal Chem 2011;401:3115–24.10.1007/s00216-011-5192-zSearch in Google Scholar PubMed
19. Jacobs M, van Greevenbroek MM, van der Kallen CJ, Ferreira I, Blaak EE, Feskens EJ, et al. Low-grade inflammation can partly explain the association between the metabolic syndrome and either coronary artery disease or severity of peripheral arterial disease: the CODAM study. Eur J Clin Invest 2009;39:437–44.10.1111/j.1365-2362.2009.02129.xSearch in Google Scholar PubMed
20. Beisswenger PJ, Howell SK, Touchette AD, Lal S, Szwergold BS. Metformin reduces systemic methylglyoxal levels in type 2 diabetes. Diabetes 1999;48:198–202.10.2337/diabetes.48.1.198Search in Google Scholar PubMed
21. Fleming T, Cuny J, Nawroth G, Djuric Z, Humpert PM, Zeier M, et al. Is diabetes an acquired disorder of reactive glucose metabolites and their intermediates? Diabetologia 2012;55:1151–5.10.1007/s00125-012-2452-1Search in Google Scholar PubMed
22. Mikesh LM, Bruns DE. Stabilization of glucose in blood specimens: mechanism of delay in fluoride inhibition of glycolysis. Clin Chem 2008;54:930–2.10.1373/clinchem.2007.102160Search in Google Scholar PubMed
23. Ohmori S, Mori M, Shiraha K, Kawase M. Biosynthesis and degradation of methylglyoxal in animals. Prog Clin Biol Res 1989;290:397–412.Search in Google Scholar
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