Original Scientific Article
Determination of aflatoxins and ochratoxin A in animal liver using HPLC-FD method with immunoaffinity column clean-up
Biljana Stojanovska-Dimzoska*,
Elizabeta Dimitrieska-Stojkovic,
Zehra Hajrulai-Musliu,
Risto Uzunov,
Aleksandra Angeleska,
Dean Jankuloski,
Katerina Blagoevska

Mac Vet Rev 2021; 44 (2): i - ix

10.2478/macvetrev-2021-0017

Received: 04 February 2021

Received in revised form: 02 April 2021

Accepted: 05 April 2021

Available Online First: 12 April 2021

Published on: 15 October 2021

Correspondence: Biljana Stojanovska-Dimzoska, bsdimzoska@fvm.ukim.edu.mk

Abstract

Analytical methods based on immunoaffinity column clean-up and quantitative determination with liquid chromatography-fluorescence detection were used to determine aflatoxins and ochratoxin A in liver samples. The validation of the procedures was performed. The linearity of the methods was checked, and a good coefficient of correlation was found for all aflatoxins and OTA as well. The LOD and LOQ were acceptable: 0.003 μg/kg and 0.009 μg/kg for AFB1; 0.001 μg/kg and 0.005 μg/kg for AFB2; 0.006 μg/kg and 0.020 μg/kg for AFG1; 0.007 μg/kg and 0.022 μg/kg for AFG2; 0.08 μg/kg and 0.27 μg/kg for OTA. The results for the repeatability estimated by the relative standard deviation (RSDr) were satisfactory and the obtained values were in the acceptable range (1.97–14.41% for all aflatoxins and 3.76-8.31% for OTA) at three proposed concentration levels. RSDR values showed acceptable correlation between two analysts for all four aflatoxins and OTA. The RSDR values were as followed: 2.37% and 5.60% for AFB1, 6.71% and 8.78% for AFB2, 4.40% and 7.00% for AFG1 and 10.30% and 13.91% for AFG2 (for the first and second analyst, respectively). The RSDR values for OTA were 4.91% and 3.15% (1 μg/kg); 3.76% and 4.12% (5 μg/kg) and 8.31% and 8.21% (10 μg/kg). The mean recovery for total aflatoxins and OTA were 78.10% and 93.34%, respectively. All validation parameters were in accordance to European legislation. They indicate that the proposed analytical procedures are suitable and they could be methods of choice for the determination of aflatoxins and OTA in liver samples.

Keywords: aflatoxins, ochratoxin A, HPLC-FD, liver samples, validation


References

  1. European Food Safety Authority (EFSA). (2006). Opinion of scientific panel on contaminants in food chain on a request from the commission related to ochratoxin A in food. EFSA J. 365, 1-56. https://doi.org/10.2903/j.efsa.2006.365
  2. European Food Safety Authority (EFSA). (2004). Opinion of scientific panel on contaminants in food chain on a request from the commission related to ochratoxin A as undesirable substance in animal feed. EFSA J. 101, 1-36. https://doi.org/10.2903/j.efsa.2004.101
  3. Binder, E.M., Tan, L.M., Chin, L.J., Handl, J., Richard, J. (2007). Worldwide occurrence of mycotoxins in commodities, feeds and feed ingredients. Anim Feed Sci Technol. 137, 265-282. https://doi.org/10.1016/j.anifeedsci.2007.06.005
  4. Scientific Cooperation (SCOOP) Task Reports. (2002). Task 3.2.7. Assessment of dietary intake of ochratoxin A by the population of EU member states. 27, Italy, 2002. http://ec.europa.ec//food/fs/scoop/3.2.7_en.pdf
  5. Sharman, M., Gilbert, J. (1991). Automated aflatoxin analysis of foods and animal feeds using immunaffinity column clean-up and high-performance liquid chromatographic determination. J Chromatogr. 543, 220-225. https://doi.org/10.1016/S0021-9673(01)95771-1
  6. Zinedine, A., Juan, C., Soriano, J.M., Moltó, J.C., Idrissi, L., Mañes, J. (2007). Limited survey for the occurrence of aflatoxins in cereals and poultry feeds from Rabat, Morocco. Int J Food Microbiol. 115, 124-127. https://doi.org/10.1016/j.ijfoodmicro.2006.10.013 PMid:17218031
  7. Ibáñez-Vea, M., Martinez, R., Gonzalez-Peñas, E., Lizarraga, E., Lopez de Cerain, A. (2011). Co-occurrence of aflatoxins, ochratoxin A and zearalenone I breakfast cereals from Spanish market. Food Control 22, 1949-1955. https://doi.org/10.1016/j.foodcont.2011.05.008
  8. Stojanovska-Dimzoska, B., Hajrulai-Musliu, Z., Dimitrieska-Stojkovic, E., Uzunov, R., Angeleska, A., Jankuloski, D. (2015). Co-occurrence of aflatoxins, ochratoxin A and zearalenon in feed components determined by liquid chromatography with fluorescence detection. 2nd International VETistanbul Group Congress April 2015, St. Petersburg, Book of Abstaracts, p. 744.
  9. Skarkova, J., Ostry, V., Malir, F., Roubal, T. (2013). Determination of ochratoxin A in food by high performance liquid chromatography. Anal Lett. 46(10): 1495-1504. https://doi.org/10.1080/00032719.2013.771266
  10. Bonerba, E., Ceci, E., Balzareti, C., Vallone, L., Crescenzo, G., Di Pinto, A., Celano, G.V., Tantillo, G., Bozzo, G. (2017). Ochratoxin A detection by HPLC-FL in processed baby foods. J Nephrol Ther. 7(4): 1000301.
  11. Corcuera, L.A., Ibáñez-Vea, M., Vettorazzi, A., Gonzalez-Peñas, E., Lopez de Cerain, A. (2011). Validation of UHPLC-FLD analytical method for the simultaneous quantification of aflatoxin B1 and ochratoxin A in rat plasma, liver and kidney. J Chromatogr. B. 879, 2733-2740. https://doi.org/10.1016/j.jchromb.2011.07.039 PMid:21868292
  12. Tavčar-Kalcher, G., Vrtač, K., Pestevšek, U., Vengušt, A. (2007). Validation of the procedure for the determination of aflatoxin B1 in animal liver using immunoaffinity columns and liquid chromatography with postcolumn derivatisation and fluorescence detection. Food Control 18, 333-337. https://doi.org/10.1016/j.foodcont.2005.10.016
  13. Kȕçȕkçakan, B., Stojanovska-Dimzoska, B., Hajrulai-Musliu, Z., Dimitrieska-Stojkovic, E., Uzunov, R., Davcheva, K. (2016). Determination of ochratoxin A in cattle lever by HPLC-FD method. Kafkas Univ Vet Fak Derg. 22(1): 1-5.
  14. Jorgensen and Petersen (2002). Content of ochratoxin A in paired kidney and meat samples from healthy Danish slaughter pigs. Food Addit Contam. 19(6): 562-567. https://doi.org/10.1080/02652030110113807 PMid:12042022
  15. Ceci, E., Bozzo, G., Bonerba, E., Di Pinto, A., Tantillo, M.G. (2007). Ochratoxin A detection by HPLC in target tissues of swine and cytological and histological analysis. Food Chem. 105, 364-368. https://doi.org/10.1016/j.foodchem.2006.12.019
  16. Commission of the European Communities. (2002). Commission Decision of 12 August 2002 implementing Council Directive 96/23/EC concerning the performance of analytical methods and the interpretation of results (2002/657/EC). Off J Eur Union. L 2218-2236.
  17. Commission of the European Communities. (2006). Commission Regulation (EC) No 401/2006 of 23 February 2006 laying down the methods of sampling and analysis for the official control of the levels of Mycotoxins in foodstuffs. Off J Eur Union. L 70, 12-34.
  18. AOAC Official Method 982.24. Aflatoxin B1 and M1 in liver (AOAC 18th Edition).
  19. ISO 16050:2003 Foodstuffs - determination of aflatoxin B1 and the total content of aflatoxins B1, B2, G1 and G2 in cereals, nuts and derived products - high-performance liquid chromatographic method.
  20. Taylor, B., Kuyatt, C. (1994). Guidelines for evaluating and expressing the uncertainty of NIST Measurement Results. In: NIST Editorial Review Boards. NIST Technical Note. 1297, 1-20. https://doi.org/10.6028/NIST.TN.1297
  21. Dragaci, S., Grosso, F.R., Bire, R., Fremy, M., Coulon, S. (1999). A French monitoring programme for determining ochratoxin A occurrence in pig kidneys. Nat Toxins. 7, 167-173. https://doi.org/10.1002/(SICI)1522-7189(199907/08)7:4<167::AID-NT55>3.0.CO;2-Q
  22. Milicevic, D., Juric, V., Stefanovic, S.M., Veskovic-Moracanin, S.V., Jankovic, S. (2009). Analysis of ochratoxin A in pig tissues using high pressure liquid chromatography (HPLC) and liquid chromatography tandem mass spectrometry as confirmative method. Proc Nat Sci Matica Srpska Novi Sad. 117, 51-61. https://doi.org/10.2298/ZMSPN0917051M
  23. Milicevic, D., Stefanovic, S., Jankovic, S., Radicevic, T. (2012). Risk analysis and exposure assessment of ochratoxin A in Serbia. Vet World. 5, 412-416. https://doi.org/10.5455/vetworld.2012.412-416


Copyright

© 2021 Stojanovska-Dimzoska B. This is an open-access article published under the terms of the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Conflict of Interest Statement

The authors have declared that no competing interests exist.

Citation Information

Macedonian Veterinary Review. Volume 44, Issue 2, Pages i-ix, e-ISSN 1857-7415, p-ISSN 1409-7621, DOI: 10.2478/macvetrev-2021-0017, 2021