Skip to main content
SpringerLink
Log in

An Improved HPLC-FLD for Fast and Simple Detection of Ethyl Carbamate in Soy Sauce and Prediction of Precursors

  • Published:
Food Analytical Methods Aims and scope Submit manuscript

Abstract

A high-performance liquid chromatography with fluorescence detection (HPLC-FLD) method for ethyl carbamate (EC) determination in Chinese soy sauce was developed and used to predict EC precursors, and extraction, derivatization, and chromatographic conditions were optimized systematically. Under optimal conditions, the limit of detection, linear range, and recoveries were 3.91 μg/L, 12.87–274.87 μg/L, and 81.5–95.4%, respectively. The method precision was less than 8.9% (RSD) and no significant difference was found between EC determinations measured by HPLC-FLD and GC/MS. Using the proposed HPLC-FLD method, the EC contents in Chinese soy sauces ranged from not detected to 64.88 μg/L (n = 19). Except for one soy sauce brand, the EC contents in low-salt solid-state fermented (LSF) soy sauces were lower than those in high-salt liquid fermented (HLF) soy sauces and were mainly affected by alcohol content. Moreover, the free ornithine and total arginine contents were significantly correlated with EC content in the soy sauce products. The results of this work provide a foundation for further study of EC formation and inhibition in Chinese soy sauces.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  • Ajtony Z, Szoboszlai N, Bencs L, Viszket E, Mihucz VG (2013) Determination of ethyl carbamate in wine by high performance liquid chromatography. Food Chem 141:1301–1305

    Article  CAS  Google Scholar 

  • Anagnostopoulos C, Miliadis G (2013) Development and validation of an easy multiresidue method for the determination of multiclass pesticide residues using GC-MS/MS and LC-MS/MS in olive oil and olives. Talanta 112:1–10

    Article  CAS  Google Scholar 

  • Bai WD, Sun SG, Zhao WH, Qian M, Liu XY, Chen WX (2016) Determination of ethyl carbamate (EC) by GC-MS and characterization of aroma compounds by HS-SPME-GC-MS during wine frying status in Hakka yellow rice wine. Food Anal Methods 10:2068–2077

    Article  Google Scholar 

  • Beland FA, Benson RW, Mellick PW, Kovatch RM, Roberts DW, Fang JL, Doerge DR (2005) Effect of ethanol on the tumorigenicity of urethane (ethyl carbamate) in B6C3F 1 mice. Food Chem Toxicol 43:1–19

    Article  CAS  Google Scholar 

  • Bruno SNF, Vaitsman DS, Kunigami CN, Brasil MG (2007) Influence of the distillation processes from Rio de Janeiro in the ethyl carbamate formation in Brazilian sugar cane spirits. Food Chem 104:1345–1352

    Article  CAS  Google Scholar 

  • Canas BJ, Havery DC, Robinson LR, Sullivan MP, Jr JF, Diachenko GW (1989) Ethyl carbamate levels in selected fermented foods and beverages. J Assoc Off Anal Chem 72:873–876

    CAS  Google Scholar 

  • Dahabieh MS, Husnik JI, Vuuren HJJV (2010) Functional enhancement of sake yeast strains to minimize the production of ethyl carbamate in sake wine. J Appl Microbiol 109:963–973

    Article  CAS  Google Scholar 

  • de Resende Machado AM, das Cardoso M, Saczk AA, dos Anjos JP, Zacaroni LM, Dórea HS, Nelson DL (2013) Determination of ethyl carbamate in cachaca produced from copper stills by HPLC. Food Chem 138:1233–1238

    Article  Google Scholar 

  • Fu ML, Liu J, Chen QH, Liu XJ, He GQ, Chen JC (2010) Determination of ethyl carbamate in Chinese yellow rice wine using high-performance liquid chromatography with fluorescence detection. Int J Food Sci Technol 45:1297–1302

    Article  CAS  Google Scholar 

  • Galinaro CA, Ohe TH, Da SA, Da SS, Franco DW (2015) Cyanate as an active precursor of ethyl carbamate formation in sugar cane spirit. J Agric Food Chem 63:7415–7420

    Article  CAS  Google Scholar 

  • Hamlet CG, Jayaratne SM, Morrison C (2005) Application of positive ion chemical ionisation and tandem mass spectrometry combined with gas chromatography to the trace level analysis of ethyl carbamate in bread. Rapid Commun Mass Spectrom 19:2235–2243

    Article  CAS  Google Scholar 

  • Hasnip S, Crews C, Potter N, Christy J, Chan D, Bondu T, Matthews W, Walters B, Patel K (2007) Survey of ethyl carbamate in fermented foods sold in the United Kingdom in 2004. J Agric Food Chem 55:2755–2759

    Article  CAS  Google Scholar 

  • Herbert P, Santos L, Bastos M, Barres P, Alves A (2002) New HPLC method to determine ethyl carbamate in alcoholic beverages using fluorescence detection. J Food Sci 67:1616–1620

    Article  CAS  Google Scholar 

  • Horii S, Goto K (2010) Determination of ethyl carbamate in sake using headspace solid phase microextraction. J Inst Brew 116:177–181

    Article  CAS  Google Scholar 

  • Koh E, Kwon H (2007) Quantification of ethyl carbamate in soy sauce consumed in Korea and estimated daily intakes by age. J Sci Food Agric 87:98–102

    Article  CAS  Google Scholar 

  • Lachenmeier DW (2005) Rapid screening for ethyl carbamate in stone-fruit spirits using FTIR spectroscopy and chemometrics. Anal Bioanal Chem 382:1407–1412

    Article  CAS  Google Scholar 

  • Lee KG (2013) Analysis and risk assessment of ethyl carbamate in various fermented foods. Eur Food Res Technol 236:891–898

    Article  CAS  Google Scholar 

  • Lee BQ, Khor SM (2015) 3-Chloropropane-1,2-diol (3-MCPD) in soy sauce: a review on the formation reduction and detection of this potential carcinogen. Compr Rev Food Sci Food Sci 14:48–66

    Article  CAS  Google Scholar 

  • Li GH, Zhong QD, Wang DB, Zhang XY, Gao HB, Shen SG (2015) Determination and formation of ethyl carbamate in Chinese spirits. Food Control 56:169–176

    Article  CAS  Google Scholar 

  • Liao QG, Luo LG (2014) Fast and selective pressurized liquid extraction with simultaneous in-cell cleanup for the analysis of ethyl carbamate in fermented solid foods. Chromatographia 77:963–967

    Article  CAS  Google Scholar 

  • Miller YE, Dwyernield LD, Keith RL, Le M, Franklin WA, Malkinson AM (2003) Induction of a high incidence of lung tumors in C57BL/6 mice with multiple ethyl carbamate injections. Cancer Lett 198:139–144

    Article  CAS  Google Scholar 

  • Park SK, Kim CT, Lee JW, Jhee OH, Om AS, Kang JS, Moon TW (2007) Analysis of ethyl carbamate in Korean soy sauce using high-performance liquid chromatography with fluorescence detection or tandem mass spectrometry and gas chromatography with mass spectrometry. Food Control 18:975–982

    Article  CAS  Google Scholar 

  • Riachi LG, Santos A, Moreira RF, De Maria CA (2014) A review of ethyl carbamate and polycyclic aromatic hydrocarbon contamination risk in cachaca and other Brazilian sugarcane spirits. Food Chem 149:159–169

    Article  CAS  Google Scholar 

  • Wang PH, Sun JY, Li XM, Wu DH, Li T, Lu J, Chen J, Xie G (2014) Contribution of citrulline to the formation of ethyl carbamate during Chinese rice wine production. Food Addit Contam 31:587–592

    Article  CAS  Google Scholar 

  • Weber JV, Sharypov VI (2009) Ethyl carbamate in foods and beverages: a review. Environ Chem Lett 7:233–247

    Article  CAS  Google Scholar 

  • Wei X, Huang Z, Zhang W, Du Y (2007) Improving the sensitivity of NIR spectroscopy with an enrichment technique: determining a trace analyte of ethyl carbamate. Anal Sci 23:853–856

    Article  CAS  Google Scholar 

  • Wu PG, Pan XD, Wang LY, Shen XH, Yang DJ (2012) A survey of ethyl carbamate in fermented foods and beverages from Zhejiang China. Food Control 23:286–288

    Article  CAS  Google Scholar 

  • Wu P, Zhang L, Shen X, Wang L, Zou Y, Zhang J, Tan Y, Tang J, Ma B, Pan X, Jiang W (2015) Determination of ethyl carbamate in Chinese yellow rice wine by diatomaceous earth extraction and GC/MS method. J AOAC Int 98:834–838

    Article  CAS  Google Scholar 

  • Yang DT, Zhou HB, Ying YB, Niessner R, Haisch C (2013) Surface-enhanced Raman scattering for quantitative detection of ethyl carbamate in alcoholic beverages. Anal Bioanal Chem 405:9419–9425

    Article  CAS  Google Scholar 

  • Yong GK, Lyu J, Kim MK, Lee KG (2015) Effect of citrulline urea ethanol and urease on the formation of ethyl carbamate in soybean paste model system. Food Chem 189:74–79

    Article  Google Scholar 

  • Zhang J, Liu GX, Zhang Y, Gao Q, Wang DP, Liu H (2014a) Simultaneous determination of ethyl carbamate and urea in alcoholic beverages by high-performance liquid chromatography coupled with fluorescence detection. J Agric Food Chem 62:2797–2802

    Article  CAS  Google Scholar 

  • Zhang JR, Fang F, Chen J, Du GC (2014b) The arginine deiminase pathway of koji bacteria is involved in ethyl carbamate precursor production in soy sauce. FEMS Microbiol Lett 358:91–97

    Article  CAS  Google Scholar 

  • Zhao XR, Jiang CX (2015) Determination of ethyl carbamate in fermented liquids by ultra high performance liquid chromatography coupled with a Q Exactive hybrid quadrupole-orbitrap mass spectrometer. Food Chem 177:66–71

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Guangdong Provincial Key Laboratory of Food Quality and Safety, College of Food Science, South China Agricultural University, Guangzhou, 510642, China

    Kai Zhou, Yao Liu, Wen-Qian Li, Yuan-Ming Sun & Zhen-Lin Xu

  2. College of Light Industry and Food Science, Zhongkai University of Agriculture and Engineering, Guangzhou, 510225, China

    Gong-Liang Liu & Wei-Dong Bai

  3. Guangdong Food Industry Research Institute, Guangzhou, 510335, China

    Na Wei

Authors
  1. Kai Zhou
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yao Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Wen-Qian Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Gong-Liang Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Na Wei
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Yuan-Ming Sun
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Wei-Dong Bai
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Zhen-Lin Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Yuan-Ming Sun or Zhen-Lin Xu.

Ethics declarations

Funding

This study was funded by the Science and Technology Foundation of Guangdong Province, China (2015B020205002, 2013B020311019, 2014TQ01N109), the National Basic Research Program of China (973 Program, 2012CB720803), the Natural Science Foundation of Guangdong Province, China (S2013030013338 and 2014A030313592), and the Technology Bureau of Zhongshan, China (2016C1013).

Conflict of Interest

Kai Zhou declares that he has no conflict of interest. Yao Liu declares that she has no conflict of interest. Wen-Qian Li declares that she has no conflict of interest. Gong-Liang Liu declares that he has no conflict of interest. Na Wei declares that she has no conflict of interest. Yuan-Ming Sun declares that he has no conflict of interest. Wei-Dong Bai declares that he has no conflict of interest. Zhen-Lin Xu declares that he has no conflict of interest.

Ethical Approval

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

Informed Consent

Not applicable.

Electronic Supplementary Material

ESM 1

(DOC 110 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Zhou, K., Liu, Y., Li, WQ. et al. An Improved HPLC-FLD for Fast and Simple Detection of Ethyl Carbamate in Soy Sauce and Prediction of Precursors. Food Anal. Methods 10, 3856–3865 (2017). https://doi.org/10.1007/s12161-017-0948-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12161-017-0948-5

Keywords

Search

Navigation