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The Simultaneous Analysis of Amadori and Heyns Compounds in Dried Fruits by High Performance Liquid Chromatography Tandem Mass Spectrometry


Amadori and Heyns compounds, which play important roles in food flavor, color, and nutritional value, are generated during the Maillard reaction. It is difficult to analyze the compounds simultaneously because the structures of Amadori and Heyns compounds are very similar. To develop a method to distinguish and analyze Amadori and Heyns compounds simultaneously by high performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS), studies of mass spectrometry fragments of the Heyns compounds obtained by electrospray ionization were performed and two characteristic fragment ion pairs of Amadori ([AA-H + CH2]+/[M + H]+) and Heyns ([M + H-2H2O–CH3OH-CO]+/[M + H]+) compounds were selected by comparing the cleavage mechanism between them. The calibration curves established with ion pairs of Fru-Pro (fructose-proline), Fru-Leu (fructose-leucine), and Fru-Val (fructose-valine) were used to calculate the content of the corresponding Amadori and Heyns compounds. The method displayed good linearity with correlation coefficients no less than 0.9989. The limit of detection was no more than 0.125 ng/mL. The relative standard deviation was less than 4.31 % and the spiked recoveries ranged from 90.55 % to 110.91 %. The method is accurate and reliable for simultaneously determining Amadori and Heyns compounds in dried raisins and jujubes and was helpful for evaluating and controlling the quality of dried fruits.

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  1. Baptista JAB, Carvalho RCB (2004) Indirect determination of Amadori compounds in milk-based products by HPLC/ELSD/UV as an index of protein deterioration. Food Res Int 37:739–747. doi:10.1016/j.foodres.2004.02.006

    CAS  Article  Google Scholar 

  2. Davidek T, Clety N, Devaud S, Robert F, Blank I (2003) Simultaneous quantitative analysis of Maillard reaction precursors and products by high performance anion exchange chromatography. J Agric Food Chem 51:7259–7265. doi:10.1021/jf034794n

    CAS  Article  Google Scholar 

  3. Davidek T, Kraehenbuehl K, Devaud S, Robert F, Blank I (2005) Analysis of Amadori compounds by high performance cation exchange chromatography coupled to tandem mass spectrometry. Anal Chem 77:140–147. doi:10.1021/ac048925a

    CAS  Article  Google Scholar 

  4. Ge S-J, Lee T-C (1996) Effective HPLC method for the determination of aromatic Amadori compounds. J Agric Food Chem 44:1053–1057

    CAS  Article  Google Scholar 

  5. Guan YG, Wang SL, Yu SJ, Yu SM, Zhao ZG (2012) Changes in the initial stages of a glucose-proline Maillard reaction model system influences dairy product quality during thermal processing. J Dairy Sci 95:590–601. doi:10.3168/jds.2011-4860

    CAS  Article  Google Scholar 

  6. Hofmann T, Schieberle P (2000) Formation of aroma-active strecker-aldehydes by a direct oxidative degradation of Amadori compounds. J Agric Food Chem 48:4301–4305

    CAS  Article  Google Scholar 

  7. Horvat S, Jakas A (2004) Peptide and amino acid glycation: new insights into the Maillard reaction. J Pept Sci 10:119–137. doi:10.1002/psc.519

    CAS  Article  Google Scholar 

  8. Hou T, Han X, Zhao J, Wang Y, Yao X, Gao H (2015) Review of grape processing technology. The Food Industry 36:223–228

    CAS  Google Scholar 

  9. Hwang IG, Kim HY, Woo KS, Lee J, Jeong HS (2011) Biological activities of Maillard reaction products (MRPs) in a sugar–amino acid model system. Food Chem 126:221–227. doi:10.1016/j.foodchem.2010.10.103

    CAS  Article  Google Scholar 

  10. Huyghues-Despointes A, Yaylayan VA(1994) A multidetector HPLC system for the analysis of Amadori and other Maiilard reaction intermediates. Food Chem 51:109–117 

    CAS  Article  Google Scholar 

  11. Jakas A, Katić A, Bionda N, Horvat Š (2008) Glycation of a lysine-containing tetrapeptide by d-glucose and d-fructose—influence of different reaction conditions on the formation of Amadori/Heyns products. Carbohydr Res 343:2475–2480. doi:10.1016/j.carres.2008.07.003

    CAS  Article  Google Scholar 

  12. Joo K-M, Park C-W, Jeong H-J, Lee SJ, Chang IS (2008) Simultaneous determination of two Amadori compounds in Korean red ginseng (Panax ginseng) extracts and rat plasma by high performance anion-exchange chromatography with pulsed amperometric detection. J Chromatogr B 865:159–166. doi:10.1016/j.jchromb.2008.02.012

    CAS  Article  Google Scholar 

  13. Krause R, Schlegel K, Schwarzer E, Henle T (2008) Formation of peptide-bound Heyns compounds. J Agric Food Chem 56:2522–2527. doi:10.1021/jf073256y

    CAS  Article  Google Scholar 

  14. Mcpherson JD, Shilton BH, Walton DJ (1988) Role of fructose in glycation and cross-linking of proteinst. Biochemistry 27:1901–1907

    CAS  Article  Google Scholar 

  15. Mossine VV, Barnes CL, Glinsky GV, Feather MS (1996) Molecular and crystal structure of N-(2-deoxy-o-aldohexos-2-yl)-glycines (Heyns compounds. Carbohydr Res 284:11–24

    CAS  Article  Google Scholar 

  16. Oh Y-C, Hartman TG, Ho C-T (1992) Volatile compounds generated from the Maillard reaction of pro-Gly, Gly-pro, and a mixture of glycine and proline with glucose. J Agric Food Chem 40:1878–1880

    CAS  Article  Google Scholar 

  17. Rivera ZS, Kennedy JF (1991) The Maillard reaction in food processing, human nutrition and physiology. Carbohydr Polym 16:460–461. doi:10.1016/0144-8617(91)90064-j

    Article  Google Scholar 

  18. Sanz ML, Del Castillo MD, Corzo N, Olano A (2001) Formation of Amadori compounds in dehydrated fruits. J Agric Food Chem 49:5228–5231. doi:10.1021/jf010580z

    CAS  Article  Google Scholar 

  19. Shipar MAH (2006) Formation of the Heyns rearrangement products in dihydroxyacetone and glycine Maillard reaction: a computational study. Food Chem 97:231–243. doi:10.1016/j.foodchem.2005.03.041

    CAS  Article  Google Scholar 

  20. Somoza V, Fogliano V (2013) 100 years of the Maillard reaction: why our food turns brown. J Agric Food Chem 61:10197–10197. doi:10.1021/jf403107k

    CAS  Article  Google Scholar 

  21. Srinivas SM, Harohally NV (2012) Improved synthesis of lysine- and arginine-derived Amadori and Heyns products and in vitro measurement of their angiotensin I-converting enzyme inhibitory activity. J Agric Food Chem 60:1522–1527. doi:10.1021/jf204185y

    CAS  Article  Google Scholar 

  22. Toda M, Heilmann M, Ilchmann A, Vieths S (2014) The Maillard reaction and food allergies: is there a link? Clin Chem Lab Med 52:61–67. doi:10.1515/cclm-2012-0830

    CAS  Article  Google Scholar 

  23. Troise AD, Fiore A, Roviello G, Monti SM, Fogliano V (2015) Simultaneous quantification of amino acids and Amadori products in foods through ion-pairing liquid chromatography–high-resolution mass spectrometry. Amino Acids 47:111–124. doi:10.1007/s00726-014-1845-5

    CAS  Article  Google Scholar 

  24. Wang J, Lu YM, Liu BZ, He HY (2008) Electrospray positive ionization tandem mass spectrometry of Amadori compounds. J Mass Spectrom 43:262–264. doi:10.1002/jms.1290

    CAS  Article  Google Scholar 

  25. Zhou WH, Wang J, Wu D (2014) Determination of important flavour precursor compounds (Amadori compounds) in cigarettes by LC-MS/MS. J Anal Chem 69:691–695

    CAS  Article  Google Scholar 

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Corresponding author

Correspondence to Jun Wang.

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Conflict of Interest

Heng Yuan declares that he has no conflict of interest. Linjuan Sun declares that he has no conflict of interest. Min Chen declares that he has no conflict of interest. Jun Wang declares that he has no conflict of interest.


This work was financially supported by the national science and technology support programs from MOST of China (2012BAD29B04–1).

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This article does not contain any studies performed by any of the authors using human participants or animals.

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Yuan, H., Sun, L., Chen, M. et al. The Simultaneous Analysis of Amadori and Heyns Compounds in Dried Fruits by High Performance Liquid Chromatography Tandem Mass Spectrometry. Food Anal. Methods 10, 1097–1105 (2017).

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  • Maillard reaction
  • Heyns compounds
  • Amadori compounds
  • Cleavage mechanism
  • Dried fruits