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Evaluation and Validation of Two Chromatographic Methods (HPLC-Fluorescence and LC–MS/MS) for the Determination and Confirmation of Ochratoxin A in Pig Tissues

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Abstract

Two different analytical methods for the determination and confirmation of ochratoxin A (OTA) in blood serum, kidney, and liver of pigs have been compared. Sample cleanup was based on liquid–liquid phase extraction. The detection of OTA was accomplished with high-performance liquid chromatography (HPLC) combined either with fluorescence detection (FLD) or electrospray ionization (ESI+) tandem mass spectrometry (MS/MS). The comparative method of evaluation was based on the investigation of 90 samples of blood serum, kidney, and liver per animal originating from different regions of Serbia. The analytical results are discussed in view of the respective method validation data and the corresponding experimental protocols. In general, analytical data obtained with liquid chromatography (LC)-MS/MS detection offered comparable good results in the sub-ppb concentration level indicating that the electrospray tandem mass spectrometric (LC–MS/MS) method was more selective and sensitive for the analysis and confirmation of OTA in pig tissues than the HPLC method after the methylation of OTA.

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References

  • Barna-Vetro I, Solti L, Teren J, Gyongyosi A, Szabo E, Wolfling A (1996) Sensitive ELISA test for determination of ochratoxin A. J Agric Food Chem 44(12):4071–4074

    Article  CAS  Google Scholar 

  • Bauer J, Gareis M, Gedek B (1984) Detection and occurrence of ochratoxin A in pigs for slaughter. Berl Muench Tieraerztl Wsch 97:279–283 (in German)

    CAS  Google Scholar 

  • Causon R (1997) Validation of chromatographic methods in biomedical analysis. Viewpoint and discussion. J Chromatogr B 689:175–180

    Article  CAS  Google Scholar 

  • Creppy EE (1999) Human ochratoxicosis. J Toxicol Toxin Rev 18(3–4):277–293

    CAS  Google Scholar 

  • Curtui VG, Gareis M (2001) A simple HPLC method for the determination of the mycotoxins ochratoxin A and B in blood serum of swine. Food Addit Contam 18(7):635–643

    CAS  Google Scholar 

  • EC (2002) Commission Directive (EC) No. 2002/26/EC of 13 March 2002. Laying down the sampling methods and the methods of analysis for the official control of the levels of ochratoxin A in foodstuffs L 75, 38

  • Eskola M, Parikka P, Rizzo A (2001) Trichothecenes, ochratoxin A and zearalenone contamination and Fusarium infection in Finnish cereal samples in 1998. Food Addit Contam 18(8):707–718

    CAS  Google Scholar 

  • Eskola M, Kokkonen M, Rizzo A (2002) Application of manual and automated Systems for purification of ochratoxin A and zearalenone in cereals with immunoaffinity columns. J Agric Food Chem 50(1):41–47

    Article  CAS  Google Scholar 

  • Gareis M (1999) Vorkommen der Mykotoxine Ochratoxin A und B in deutscher Braugerste und daraus hergestelltem Malz. Arch Lebensmittelhyg 50:83–87

    CAS  Google Scholar 

  • IARC (International Agency for Research on Cancer) (1993) Monographs on the evaluation of carcinogenic risks to humans, vol 56. IARC, Geneva

    Google Scholar 

  • JECFA (Joint FAO/WHO Expert Committee on Food Additives) (2001) Toxicological evaluation of certain food additives. In Proceedings of the fifty-sixth meeting of JECFA. WHO Food Additives Series. World Health Organization, Geneva, Switzerland

  • Jorgensen K (1998) Survey of pork, poultry, coffee, beer and pulses for ochratoxin A. Food Addit Contam 15:550–554

    CAS  Google Scholar 

  • Lau BPY, Scott PM, Lewis DA, Kanhere SR (2000) Quantitative determination of ochratoxin A by liquid chromatography/electrospray tandem mass spectrometry. J Mass Spectrom 35:23–32

    Article  CAS  Google Scholar 

  • Losito I, Monaci L, Palmisano F, Tantillo G (2004) Determination of ochratoxin A in meat products by high-performance liquid chromatography coupled to electrospray ionisation sequential mass spectrometry. Rapid Commun Mass Spectrom 18:1965–1971

    Article  CAS  Google Scholar 

  • Matrella R, Monaci L, Milillo MA, Palmisano F, Tantillo MG (2006) Ochratoxin A determination in paired kidneys and muscle samples from swines slaughtered in southern Italy. Food Contam 117:14–117

    Google Scholar 

  • Milićević D, Jurić V, Mandić M, Đorđević M (2007) The presence of ochratoxin A residue in blood plasma of slaughtered swine. Matica Srpska Proc Nat Sci 113:55–62

    Google Scholar 

  • Milićević D, Verica Jurić, Stefanović S, Jovanović M, Janković S (2008) Survey of slaughtered pigs for occurrence of ochratoxin A and porcine nephropathy in Serbia. Int J Mol Sci 9:2169–2183

    Article  Google Scholar 

  • Miller JD, Trenholm HL (eds) (1994) Mycotoxins in grain: compounds other than aflatoxin. Eagan Press, St. Paul, MN

    Google Scholar 

  • Miraglia M, De-Dominicis A, Brera C, Corneli S, Cava E, Menghetti E, Miraglia E (1995) Ochratoxin A levels in human milk and related food samples: an exposure assessment. Nat Toxins 3:436–444

    Article  CAS  Google Scholar 

  • Monaci L, Tantillo G, Palmisano F (2004) Determination of ochratoxin A in pig tissues by liquid–liquid extraction and clean-up and high-performance liquid chromatography. Anal Bioanal Chem 378:1777–1782

    Article  CAS  Google Scholar 

  • Pittet A (2001) Natural occurrence of mycotoxins in foods and feeds: a decade review. In: de Koe WJ, Samson RA, van Egmond HP, Gilbert J, Sabino M (eds) Mycotoxins and phycotoxins in perspective at the turn of the millennium. Proceedings of the Xth international IUPAC symposium on mycotoxins and phycotoxins. Wageningen Academic Publishers, Wageningen, The Netherlands, pp 153–172

  • Richard JL, Plattner RD, May J, Liska SL (1999) The occurrence of ochratoxin A in dust collected from a problem household. Mycopathologia 146:99–103

    Article  CAS  Google Scholar 

  • Santos EA, Vargas EA (2002) Immunoaffinity column clean-up and thin layer chromatography for determination of ochratoxin A in green coffee. Food Addit Contam 19(5):447–458

    Article  CAS  Google Scholar 

  • Valenta H (1998) Chromatographic methods for the determination of ochratoxin A in animal and human tissues and fluids. J Chromatogr A 815:75–92

    Article  CAS  Google Scholar 

  • Van Egmond HP (1995) Mycotoxins: regulations, quality assurance and reference materials. Food Addit Contam 12:321–330

    Google Scholar 

  • Visconti A, Pascale M, Centonze G (1999) Determination of ochratoxin A in wine by means of immunoaffinity column clean-up and high-performance liquid chromatography. J Chromatogr A 864:89–101

    Article  CAS  Google Scholar 

  • Weidenbörner M (2001) Encyclopedia of food mycotoxins. Springer, Berlin

    Google Scholar 

  • WHO/FAO (1995) Evaluation of certain food additives and contaminants In: The 37th Report of the Joint FAO/WHO Expert Committee on Food Additives. WHO Technical Report Series No. 859. WHO, Geneva

  • Wood GM, Entwisle AC, Patel S, Hald B, Boenke A (1995) Ochratoxin A in cereals and the BCR-M&T-Projects. Nat Toxins 3:275–279

    Article  CAS  Google Scholar 

  • Zimmerli B, Dick R (1995) Determination of ochratoxin A at the ppt level in human blood, serum, milk and some foodstuffs by high-performance liquid chromatography with enhanced fluorescence detection and immunoaffinity column cleanup methodology and Swiss data. J Chromatogr B 666:85–99

    Article  CAS  Google Scholar 

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Acknowledgment

This study was funded by the Ministry of Science and Technological development, Belgrade, Serbia (project code TP-20207A). We are grateful to Ministry for their understanding and support to veterinary development.

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Authors and Affiliations

  1. Institute of Meat Hygiene and Technology, Kaćanskog 13, 11000, Belgrade, Serbia

    Dragan Milićević, Srđan Stefanović, Tatjana Baltić & Saša Janković

  2. Department for Animal Sciences, Faculty of Agriculture, University of Novi Sad, Trg Dositeja Obradovića 10, 21000, Novi Sad, Serbia

    Verica Jurić

Authors
  1. Dragan Milićević
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  2. Verica Jurić
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  3. Srđan Stefanović
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  4. Tatjana Baltić
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  5. Saša Janković
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Correspondence to Dragan Milićević.

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Milićević, D., Jurić, V., Stefanović, S. et al. Evaluation and Validation of Two Chromatographic Methods (HPLC-Fluorescence and LC–MS/MS) for the Determination and Confirmation of Ochratoxin A in Pig Tissues. Arch Environ Contam Toxicol 58, 1074–1081 (2010). https://doi.org/10.1007/s00244-009-9436-2

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  • DOI: https://doi.org/10.1007/s00244-009-9436-2

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