Skip to content
Licensed Unlicensed Requires Authentication Published by De Gruyter August 5, 2019

Measurement of α-dicarbonyl compounds in human saliva by pre-column derivatization HPLC

Xin-Jie Wang, Hong-Xia Zhang, Heng Li, Ai-Hua Zhu and Wen-Yun Gao

Abstract

Background

α-Dicarbonyl compounds (α-DCs) have been detected in body fluids including plasma and urine and elevation of this sort of compounds in vivo has been associated with the development of many kinds of chronic diseases. However whether α-DCs are present in human saliva, and if their presence/absence can be related with various chronic diseases is yet to be determined.

Methods

In this study, a pre-column derivatization HPLC-UV method was developed to measure 3-deoxyglucosone (3-DG), glyoxal (GO), methylglyoxal (MGO), diacetyl (DA), and pentane-2,3-dione (PD) in human saliva employing 4-(2,3-dimethyl-6-quinoxalinyl)-1,2-benzenediamine (DQB) as a derivatizing reagent. The derivatization of the α-DCs is fast and the conditions are facile. The method was evaluated and the results show that it is suitable for the quantification of α-DCs in human saliva.

Results

In the measurements of these α-DCs in the saliva of 15 healthy subjects and 23 type 2 diabetes mellitus (T2DM) patients, we found that the concentrations of GO and MGO in the saliva of the diabetic patients were significantly higher than those in healthy subjects. As far as we know, this is the first time that salivary α-DC concentrations have been determined and associated with T2DM.

Conclusions

The developed method would be useful for the measurement of the salivary α-DC levels and the data acquired could be informative in the early screening for diabetes.

  1. Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.

  2. Research funding: This work was supported by the Natural Science Foundation of China (Grants 21402152, 31770848), the Northwest University graduate independent innovation project (No. YZZ17154), and the Scientific Research Project of Shaanxi Province (No. 2017JM2027).

  3. Employment or leadership: None declared.

  4. Honorarium: None declared.

  5. Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.

References

1. Brings S, Fleming T, Freichel M, Muckenthaler MU, Herzig S, Nawroth PP. Dicarbonyls and advanced glycation end-products in the development of diabetic complications and targets for intervention. Int J Mol Sci 2017;18:984.10.3390/ijms18050984Search in Google Scholar PubMed PubMed Central

2. Henning C, Liehr K, Girndt M, Ulrich C, Glomb MA. Extending the spectrum of α-dicarbonyl compounds in vivo. J Biol Chem 2014;289:28676–88.10.1074/jbc.M114.563593Search in Google Scholar PubMed PubMed Central

3. 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

4. Bilova T, Paudel G, Shilyaev N, Schmidt R, Brauch D, Tarakhovskaya E, et al. Global proteomic analysis of advanced glycation end products in the Arabidopsis proteome provides evidence for age-related glycation hotspots. J Biol Chem 2017;292:15758–76.10.1074/jbc.M117.794537Search in Google Scholar PubMed PubMed Central

5. Gobert J, Glomb MA. Degradation of glucose: reinvestigation of reactive α-dicarbonyl compounds. J Agric Food Chem 2009;57:8591–7.10.1021/jf9019085Search in Google Scholar PubMed

6. 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

7. Rabbani N, Thornalley PJ. Measurement of methylglyoxal by stable isotopic dilution analysis LC-MS/MS with corroborative prediction in physiological samples. Nat Protoc 2014;9:1969–79.10.1038/nprot.2014.129Search in Google Scholar PubMed

8. Scheijen JL, Schalkwijk CG. Quantification of glyoxal, methylglyoxal and 3-deoxyglucosone in blood and plasma by ultra performance liquid chromatography tandem mass spectrometry: evaluation of blood specimen. Clin Chem Lab Med 2014;52:85–91.10.1515/cclm-2012-0878Search in Google Scholar PubMed

9. Ogasawara Y, Tanaka R, Koike S, Horiuchi Y, Miyashita M, Arai M. Determination of methylglyoxal in human blood plasma using fluorescence high performance liquid chromatography after derivatization with 1,2-diamino-4,5-methylenedioxybenzene. J Chromatogr B 2016;1029–1030:102–5.10.1016/j.jchromb.2016.07.019Search in Google Scholar PubMed

10. Wang XJ, Ma SB, Liu ZF, Li H, Gao WY. Elevated levels of α-dicarbonyl compounds in the plasma of type II diabetics and their relevance with diabetic nephropathy. J Chromatogr B 2019;1106–1107:19–25.10.1016/j.jchromb.2018.12.027Search in Google Scholar PubMed

11. Hurtado-Sánchez Mdel C, Espinosa-Mansilla A, Rodríguez-Cáceres MI, Martín-Tornero E, Durán-Merás I. Development of a method for the determination of advanced glycation end products precursors by liquid chromatography and its application in human urine samples. J Sep Sci 2012;35:2575–84.10.1002/jssc.201200255Search in Google Scholar PubMed

12. Espinosa-Mansilla A, Durán-Merás I, Cañada FC, Márquez MP. High-performance liquid chromatographic determination of glyoxal and methylglyoxal in urine by prederivatization to lumazinic rings using in serial fast scan fluorimetric and diode array detectors. Anal Biochem 2007;371:82–91.10.1016/j.ab.2007.07.028Search in Google Scholar PubMed

13. Mittelmaier S, Fünfrocken 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

14. Buczko P, Zalewska A, Szarmach I. Saliva and oxidative stress in oral cavity and in some systemic disorders. J Physiol Pharmacol 2015;63:3–9.Search in Google Scholar

15. Malamud D. Saliva as a diagnostic fluid. Dent Clin North Am 2011;55:159–78.10.1016/j.cden.2010.08.004Search in Google Scholar PubMed PubMed Central

16. Kaufman E, Lamster IB. The diagnostic applications of saliva – a review. Crit Rev Oral Biol Med 2002;13:197–212.10.1177/154411130201300209Search in Google Scholar PubMed

17. Dator R, Carrà A, Maertens L, Guidolin V, Villalta PW, Balbo S. A high resolution/accurate mass (HRAM) data-dependent MS3 neutral loss screening, classification, and relative quantitation methodology for carbonyl compounds in saliva. J Am Soc Mass Spectrom 2017;28:608–18.10.1007/s13361-016-1521-ySearch in Google Scholar PubMed PubMed Central

18. Wang XJ, Wang JY, Li H, Hui X, Gao WY. Determination of diacetyl in liquors by high performance liquid chromatography coupled with precolumn derivatization using 3,3′-diaminobenzidine. Chin J Chromatogr 2017;35:837–42.10.3724/SP.J.1123.2017.04031Search in Google Scholar PubMed

19. Wang JY, Wang XJ, Hui X, Hua SH, Li H, Gao WY. Determination of diacetyl in beer by a pre-column derivatization-HPLC-UV method using 4-(2,3-dimethyl-6-quinoxalinyl)-1,2-benzenediamine as a derivatizing reagent. J Agric Food Chem 2017;65:2635–41.10.1021/acs.jafc.7b00990Search in Google Scholar PubMed

20. Wang XJ, Gao F, Li LC, Hui X, Li H, Gao WY. Quantitative analyses of α-dicarbonyl compounds in food samples by HPLC using 4-(2,3-dimethyl-6-quinoxalinyl)-1,2-benzenediamine as a derivatizing reagent. Microchem J 2018;141:64–70.10.1016/j.microc.2018.05.006Search in Google Scholar

21. Herold DA, Boyd JC, Bruns DE, Emerson JC, Burns KG, Bray RE, et al. Measurement of glycosylated hemoglobins using boronate affinity chromatography. Ann Clin Lab Sci 1983;13:482–8.Search in Google Scholar

22. Raabo E, Terkildsen TC. On the enzymatic determination of blood glucose. Scand J Clin Lab Invest 1960;12:402–7.10.3109/00365516009065404Search in Google Scholar PubMed

23. Nagler RM. Saliva as a tool for oral cancer diagnosis and prognosis. Oral Oncol 2009;45:1006–10.10.1016/j.oraloncology.2009.07.005Search in Google Scholar

24. Niwa T. 3-Deoxyglucosone: metabolism, analysis, biological activity, and clinical implication. J Chromatogr B Biomed Sci Appl 1999;731:23–36.10.1016/S0378-4347(99)00113-9Search in Google Scholar

25. Khuhawar MY, Kandhro AJ, Khand FD. Liquid chromatographic determination of glyoxal and methylglyoxal from serum of diabetic patients using meso-stilbenediamine as derivatizing reagent. Anal Lett 2006;39:2205–15.10.1080/00032710600754093Search in Google Scholar

26. 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

27. 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

28. 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

29. Wang B, Du J, Zhu Z, Ma Z, Wang S, Shan Z. Evaluation of parotid salivary glucose level for clinical diagnosis and monitoring type 2 diabetes mellitus patients. Biomed Res Int 2017;2017:1–5.10.1155/2017/2569707Search in Google Scholar PubMed PubMed Central

30. Darwazeh AM, MacFarlane TW, McCuish A, Lamey PJ. Mixed salivary glucose levels and candidal carriage in patients with diabetes mellitus. J Oral Pathol Med 2010;20:280–3.10.1111/j.1600-0714.1991.tb00928.xSearch in Google Scholar PubMed

31. Lima-Aragão MV, de Oliveira-Junior Jde J, Maciel MC, Silva LA, do Nascimento FR, Guerra RN. Salivary profile in diabetic patients: biochemical and immunological evaluation. BMC Res Notes 2016;9:103/1–103/7.10.1186/s13104-016-1881-1Search in Google Scholar PubMed PubMed Central

32. Gupta S, Sandhu SV, Bansal H, Sharma D. Comparison of salivary and serum glucose levels in diabetic patients. J Diabetes Sci Technol 2015;9:91–6.10.1177/1932296814552673Search in Google Scholar PubMed PubMed Central

33. Mascarenhas P, Fatela B, Barahona I. Effect of diabetes mellitus type 2 on salivary glucose – a systematic review and meta-analysis of observational studies. PLoS One 2014;9:e101706.10.1371/journal.pone.0101706Search in Google Scholar PubMed PubMed Central

34. Kaufman E, Lamster IB. The diagnostic applications of saliva – a review. Crit Rev Oral Biol Med 2002;1:197–212.10.1177/154411130201300209Search in Google Scholar PubMed

35. Shangari N, Mehta R, O’brien PJ. Hepatocyte susceptibility to glyoxal is dependent on cell thiamin content. Chem Biol Interact 2007;165:146–54.10.1016/j.cbi.2006.11.009Search in Google Scholar PubMed

36. Förster A, Kühne Y, Henle T. Studies on absorption and elimination of dietary Maillard reaction products. Ann N Y Acad 2005;1043:474–81.10.1196/annals.1333.054Search in Google Scholar PubMed


Supplementary Material

The online version of this article offers supplementary material (https://doi.org/10.1515/cclm-2019-0350).


Received: 2019-01-02
Accepted: 2019-06-17
Published Online: 2019-08-05
Published in Print: 2019-11-26

© 2019 Walter de Gruyter GmbH, Berlin/Boston

Scroll Up Arrow