Food Analytical Methods

, Volume 6, Issue 4, pp 1024–1032 | Cite as

Validation of an HPLC Methodology for the Identification and Quantification of Biogenic Amines in Chicken Meat

  • César A. Lázaro
  • Carlos A. Conte-Júnior
  • Fernanda L. Cunha
  • Eliane T. Mársico
  • Sérgio B. Mano
  • Robson M. Franco


This study validated a high performance liquid chromatography (HPLC) method to determine biogenic amines in chicken meat. For the identification of biogenic amines, an isocratic elution system coupled with a UV detector (254 nm) was used after a perchloric acid (5 %) extraction and benzoyl chloride derivatization of the samples. The standards of tyramine, putrescine, cadaverine, spermidine, and spermine were used for the following validation parameters: selectivity, linearity, accuracy, recovery, limit of detection and quantification, and robustness. Finally, chicken meat commercialized in two types of packaging was evaluated. The results showed an excellent selectivity and separation of all amines, r 2 > 0.99, relative standard deviation <5 %, recovery between 64 and 112 %, limits of detection and quantification between 0.03–1.25 and 0.15–5.00 μg L−1, respectively, and appropriate robustness for the proposed methodology. Moreover, both chicken meat commercial packages had similar values for all amines; only tyramine was significantly different (P ≤ 0.05). The proposed method was suitable to detection and quantification of simultaneous five biogenic amines in chicken meat.


Validation HPLC Biogenic amines Meat Chicken 



César Lázaro acknowledges the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior for the scholarship Programa Estudante Convenio Pos-Graduação.


  1. ABEF (2011) Brazilian chicken producer and exporters association. Annual report 2010/2011. In: Brazilian Poultry AssociationGoogle Scholar
  2. ANVISA (2003) Agência Nacional de Vigilância Sanitária - RE N° 899. Guia para Validação de Métodos Analíticos e Bioanalíticos. Diário Oficial da União, BrasiliaGoogle Scholar
  3. AOAC (2002) Guidelines for single laboratory validation of chemical methods for dietary supplements and botanicals. In: AOAC InternationalGoogle Scholar
  4. Balamatsia CC, Paleologos EK, Kontominas MG, Savvaidis IN (2006) Correlation between microbial flora, sensory changes and biogenic amines formation in fresh chicken meat stored aerobically or under modified atmosphere packaging at 4 degrees C: possible role of biogenic amines as spoilage indicators. Anton Leeuw Int J G 89(1):9–17CrossRefGoogle Scholar
  5. Baston O, Stroia AL, Moise D, Barna O (2008) Validation study of a HPLC method able to measure biogenic amines in chicken meat. Food Technol 31:44–50Google Scholar
  6. Buňková L, Buňka F, Klčovská P, Mrkvička V, Doležalová M, Kráčmar S (2010) Formation of biogenic amines by Gram-negative bacteria isolated from poultry skin. Food Chem 121(1):203–206. doi: 10.1016/j.foodchem.2009.12.012 CrossRefGoogle Scholar
  7. Dadáková E, Křížek M, Pelikánová T (2009) Determination of biogenic amines in foods using ultra-performance liquid chromatography (UPLC). Food Chem 116(1):365–370CrossRefGoogle Scholar
  8. EFSA (2011) Scientific Opinion on risk based control of biogenic amine formation in fermented foods. EFSA Journal 9(10):2393Google Scholar
  9. EURACHEM (1998) The fitness for purpose of analytical methods: a laboratory guide to method validation and related topics.45Google Scholar
  10. Gallas L, Standarová E, Steinhauserová I, Steinhauser L, Vorlová L (2010) Formation of biogenic amines in chicken meat stored under modified atmosphere. Acta Vet Brno 29:107–116CrossRefGoogle Scholar
  11. GraphPad (2007) GraphPad Prisma Version 5.0 GraphPad Software Inc San Diego, CA, USAGoogle Scholar
  12. Halász A, Baráth Á, Simon-Sarkadi L, Holzapfel W (1994) Biogenic amines and their production by microorganisms in food. Trends Food Sci Technol 5(2):42–49CrossRefGoogle Scholar
  13. ICH (1995) Validation of Analytical Procedures: Text and Methodology ICH guidelines Topic Q2 (R1). London, UKGoogle Scholar
  14. INMETRO (2011) Orientações sobre Validação de Métodos Analiticos, DOQ-CGCRE-008. Instituto Nacional de Metrologia, Normalização e Qualidade Industrial BrasilGoogle Scholar
  15. ISO (1999) IEC 17025 General Requirements for the Competence of Testing and Calibration Laboratories. In: ISO (ed). GenevaGoogle Scholar
  16. Kozová M, Kalač P, Pelikánová T (2009) Contents of biologically active polyamines in chicken meat, liver, heart and skin after slaughter and their changes during meat storage and cooking. Food Chem 116(2):419–425CrossRefGoogle Scholar
  17. Özdestan Ö, Üren A (2009) A method for benzoyl chloride derivatization of biogenic amines for high performance liquid chromatography. Talanta 78(4–5):1321–1326CrossRefGoogle Scholar
  18. Paschoal JAR, Rath S, Airoldi FPS, Reyes FGR (2008) Validação de métodos cromatográficos para a determinação de resíduos de medicamentos veterinários em alimentos. Quim Nova 31:1190–1198CrossRefGoogle Scholar
  19. Ribani M, Bottoli CBG, Collins CH, Jardim ICSF, Melo LFC (2004) Validação em métodos cromatográficos e eletroforéticos. Quim Nova 27:771–780CrossRefGoogle Scholar
  20. Rokka M, Eerola S, Smolander M, Alakomi H-L, Ahvenainen R (2004) Monitoring of the quality of modified atmosphere packaged broiler chicken cuts stored in different temperature conditions: B. Biogenic amines as quality-indicating metabolites. Food Control 15(8):601–607CrossRefGoogle Scholar
  21. Ruiz-Capillas C, Jiménez-Colmenero F (2004) Biogenic amines in meat and meat products. Crit Rev Food Sci Nutr 44(7/8):489–499Google Scholar
  22. Sander JE, Cai T, Dale N, Bennett LW (1996) Development of biogenic amines during fermentation of poultry carcasses. J Appl Poultry Res 5(2):161–166Google Scholar
  23. Shah V, Midha K, Findlay J et al (2000) Bioanalytical method validation—a revisit with a decade of progress. Pharm Res 17(12):1551–1557CrossRefGoogle Scholar
  24. Silva CMG, Gloria MBA (2002) Bioactive amines in chicken breast and thigh after slaughter and during storage at 4+/−1 oC and in chicken-based meat products. Food Chem 78(2):241–248CrossRefGoogle Scholar
  25. Tamim NM, Doerr JA (2003) Effect of putrefaction of poultry carcasses prior to rendering on biogenic amine production. J Appl Poultry Res 12(4):456–460Google Scholar
  26. Vinci G, Antonelli ML (2002) Biogenic amines: quality index of freshness in red and white meat. Food Control 13(8):519–524CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • César A. Lázaro
    • 1
  • Carlos A. Conte-Júnior
    • 2
  • Fernanda L. Cunha
    • 1
  • Eliane T. Mársico
    • 2
  • Sérgio B. Mano
    • 2
  • Robson M. Franco
    • 2
  1. 1.Postgraduate Program in Veterinary MedicineUniversidade Federal FluminenseNiteróiBrazil
  2. 2.Department of Food Technology, Faculty of VeterinaryUniversidade Federal FluminenseNiteróiBrazil

Personalised recommendations