Advertisement

Mycotoxin Research

, Volume 31, Issue 2, pp 109–115 | Cite as

A new approach using micro HPLC-MS/MS for multi-mycotoxin analysis in maize samples

  • Sebastian Hickert
  • Johannes Gerding
  • Edson Ncube
  • Florian Hübner
  • Bradley Flett
  • Benedikt Cramer
  • Hans-Ulrich Humpf
Original Paper

Abstract

Using micro high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) a simple and fast method for the quantitative determination of 26 mycotoxins was developed. Sample preparation consists of a single extraction step and a dilute-and-shoot approach without further cleanup. With a total run time of 9 min and solvent consumption below 0.3 mL per chromatographic run, the presented method is cost-effective. All toxins regulated by the European Commission with maximum or guidance levels in grain products (fumonisins B1 and B2 (FB1 and FB2)); deoxynivalenol (DON); aflatoxins B1, G1, B2, and G2 (AFB1, AFG1, AFB2, and AFG2); ochratoxin A (OTA); T-2 and HT-2 toxins; and zearalenone (ZEN) can be quantified with this method. Furthermore, the enniatins B, B1, A, and A1 (EnB, EnB1, EnA, and EnA1); beauvericin (BEA); 3-acetyl-deoxynivalenol (3-AcDON); fusarin C (FusC); sterigmatocystin (STC); gliotoxin (GT); and the Alternaria toxins alternariol (AOH), alternariol monomethyl ether (AME), altenuene (ALT), tentoxin (TEN), and altertoxin I (ATX I) can also be quantified. For all regulated compounds, recoveries ranged between 76 and 120 %. For all other toxins, the recovery was at least 51 %. The method was applied for the analysis of 42 maize samples from field trials in South Africa.

Keywords

Micro LC Mass spectrometry Capillary chromatography Aflatoxin Multi-method 

Notes

Acknowledgments

We thank AB SCIEX (Darmstadt) for supplying us with a QTRAP 5500 mass spectrometer.

Source of funding

We thank the NRW Graduate School of Chemistry for the financial support. Part of this work is based on the research supported by the National Research Foundation of South Africa, Grant Number 84162.

Conflict of interest

None

Supplementary material

12550_2015_221_MOESM1_ESM.pdf (2.2 mb)
ESM 1 (PDF 2211 kb)

References

  1. Beyer M, Ferse I, Humpf H-U (2009) Large scale production of selected type a trichothecenes: the Use of HT-2 toxin and T-2 triol as precursors for the synthesis of d3-T-2 and d3-HT-2 toxin. Mycotox Res 25:41–52CrossRefGoogle Scholar
  2. Bretz M, Beyer M, Cramer B, Humpf H-U (2006) Stable isotope dilution analysis of the Fusarium mycotoxins deoxynivalenol and 3-acetyldeoxynivalenol. Mol Nutr Food Res 50:251–260CrossRefPubMedGoogle Scholar
  3. Codex Alimentarius Committee (CAC) (1995) Codex general standard for contaminants and toxins in food and feed. (http://www.fao.org/fileadmin/user_upload/agns/pdf/CXS_193e.pdf)
  4. Codex Alimentarius Committee (CAC) (2003) Code of practice for the prevention and reduction of mycotoxin contamination in cereals, including annexes on ochratoxin a, zearalenone, fumonisins and trichothecenes, 2003 http://www.codexalimentarius.org/input/download/standards/406/CXP_051e.pdf
  5. Cramer B, Bretz M, Humpf H-U (2007) Stable isotope dilution analysis of the Fusarium mycotoxin zearalenone. J Agr Food Chem 21:8353–8358CrossRefGoogle Scholar
  6. Cramer B, Königs M, Humpf H-U (2008) Identification and in vitro cytotoxicity of ochratoxin a degradation products formed during coffee roasting. J Agr Food Chem 56:5673–5681CrossRefGoogle Scholar
  7. Department of Health, Government of South Africa (DoH) (2004) Notice No. 26849. Regulations governing tolerance for fungus-produced toxins in foodstuffs. Regulation for Act No. 54 of 1972Google Scholar
  8. European Commission Recommendation (EC) (2013) EC 2013/165/EU on the presence of T-2 and HT-2 toxin in cereals and cereal products. O J L91, 12-15Google Scholar
  9. European Commission Regulation (EC) (2006) EC 401/2003 Laying down the methods of sampling and analysis for the official control of the levels of mycotoxins in foodstuffs. O J L70, 12-34Google Scholar
  10. European Commission Regulation (EC) (2006) EC 1881/2006 Setting maximum levels for certain contaminants in foodstuffs. O J L364, 15-17Google Scholar
  11. European Committee for Standardization (CEN) (2010) Food analysis―performance criteria for single laboratory validated methods of analysis for the determination of mycotoxins (CEN/TR 16059:2010). Austrian Standards Institute, ViennaGoogle Scholar
  12. European Food Safety Authority (EFSA) (2011) Scientific opinion on the risks for animal and public health related to the presence of Alternaria toxins in feed and food. EFSA J 9(10):2407, 1-97Google Scholar
  13. European Food Safety Authority (EFSA) (2013) Scientific opinion on the risk for public and animal health related to the presence of sterigmatocystin in food and feed. EFSA J 11(6):3254, 1-81Google Scholar
  14. European Food Safety Authority (EFSA) (2014) Request for an EFSA opinion on the risks to human and animal health related to the presence of beauvericin and enniatins in food and feed. (http://registerofquestions.efsa.europa.eu/raw-war/?wicket:interface=:4)
  15. Frenich AG, Vidal JLM, del Romero-González R, Mar Aguilera-Luiz M (2009) Simple and high-throughput method for multimycotoxin analysis in cereals and related foods by ultra-high performance liquid chromatography/tandem mass spectrometry. Food Chem 117:705–712CrossRefGoogle Scholar
  16. Goryacheva IY, De Saeger S, Eremin SA, Peteghem C (2007) Immunochemical methods for rapid mycotoxin detection: evolution from single to multi analyte screening: a review. Food Addit Contam 24:1169–1183CrossRefPubMedGoogle Scholar
  17. Goswami RS, Kistler HC (2004) Heading for disaster: Fusarium graminearum on cereal crops. Mol Plant Pathol 5(6):515–525CrossRefPubMedGoogle Scholar
  18. Herzallah SM (2009) Determination of aflatoxins in eggs, milk, meat and meat products using HPLC fluorescent and UV detectors. Food Chem 114:1141–1146CrossRefGoogle Scholar
  19. Hübner F, Harrer H, Fraske A, Kneifel S, Humpf H-U (2012) Large scale purification of B-type fumonisins using centrifugal partition chromatography (CPC). Mycotox Res 28:37–43CrossRefGoogle Scholar
  20. Ishii D, Asai K, Hibi K, Jonokuchi T, Nagaya M (1977) A study of micro-high-performance liquid chromatography I development of technique for miniaturization of high-performance liquid chromatography. J Chromatogr 144:157–168CrossRefGoogle Scholar
  21. Kleigrewe K, Niehaus E-M, Wiemann P, Tudzynski B, Humpf H-U (2012) New approach via gene knockout and single-step chemical reaction for the synthesis of isotopically labeled fusarin C as an internal standard for the analysis of this Fusarium mycotoxin in food and feed samples. J Agr Food Chem 60(34):8350–8355CrossRefGoogle Scholar
  22. Legido-Quigley C, Smith NW, Mallet D (2002) Quantification of the sensitivity increase of a micro-high-performance liquid chromatography-electrospray ionization mass spectrometry system with decreasing column diameter. J Chromatogr A 976:11–18CrossRefPubMedGoogle Scholar
  23. Lukacs Z, Schaper S, Herderich M, Schreier P, Humpf H-U (1996) Identification and determination of fumonisin FB1 and FB2 in corn and corn products by high-performance liquid chromatography-electrospray-ionization tandem mass spectrometry (HPLC-ESI-MS-MS). Chromatographia 43:124–128CrossRefGoogle Scholar
  24. Monbaliu S, Poucke CV, Detavernier C, Dumoulin F, Van de Velde M, Schoeters E, Van Dyck S, Averkieva O, Van Peteghem C, De Saeger S (2010) Occurrence of mycotoxins in feed as analyzed by a multi-mycotoxin LC-MS/MS method. Food Chem 58:66–71CrossRefGoogle Scholar
  25. Oueslati S, Romero-González R, Lasram S, Frenich AG, Vidal JLM (2012) Multi-mycotoxin determination in cereals and derived products marketed in Tunisia using ultra-high performance liquid chromatography coupled to triple quadrupole mass spectrometry. Food Chem Toxicol 50:2376–2381CrossRefPubMedGoogle Scholar
  26. Pearson SM, Candish AAG, Aidoo KE, Smith JE (1999) Determination of aflatoxin levels in pistachio and cashew nuts using immunoaffinity column clean-Up with HPLC and fluorescence detection. Biotechnol Tech 13:97–99CrossRefGoogle Scholar
  27. Rasmussen RR, Storm IMLD, Rasmussen PH, Smedsgaard J, Nielsen KF (2010) Multi-mycotoxin analysis of maize silage by LC-MS/MS. Anal Bioanal Chem 397:765–776CrossRefPubMedGoogle Scholar
  28. Ren Y, Zhang Y, Shao S, Cai Z, Feng L, Wang Z (2007) Simultaneous determination of multi-component mycotoxins residues in foods and feeds by ultra performance liquid chromatography tandem mass spectrometry. J Chromatogr A 1143:48–64CrossRefPubMedGoogle Scholar
  29. Soriano JM, Gragacci S (2004) Occurrence of fumonisins in foods. Food Res Int 37:985–1000CrossRefGoogle Scholar
  30. Spanjer MC, Rensen PM, Scholten JM (2008) LC-MS/MS multi-method for mycotoxins after single extraction, with validation data for peanut, pistachio, wheat, maize, cornflakes, raisins and figs. Food Addit Contam 25(4):472–489CrossRefGoogle Scholar
  31. Sulyok M, Berthiller F, Krska R, Schuhmacher R (2006) Development and validation of a liquid chromatography/tandem mass spectrometric method for the determination of 39 mycotoxins in wheat and maize. Rapid Commun Mass Sp 20:2649–2659CrossRefGoogle Scholar
  32. Sulyok M, Krska R, Schuhmacher R (2007) A liquid chromatography/tandem mass spectrometric multi-mycotoxin method for the quantification of 87 analytes and its application to semi-quantitative screening of moldy food samples. Anal Bioanal Chem 389:1505–1523CrossRefPubMedGoogle Scholar
  33. Sulyok M, Krska R, Schuhmacher R (2010) Application of an LC-MS/MS based multi-mycotoxin method for the semi-quantitative determination of mycotoxins occurring in different types of food infected by moulds. Food Chem 119:408–416CrossRefGoogle Scholar
  34. Takeuchi T, Ishii D (1980) Ultra-micro high-performance liquid chromatography. J Chromatogr 190:150–155CrossRefGoogle Scholar
  35. Tanaka T, Yoneda A, Inoue S, Sugiura Y, Ueno Y (2000) Simultaneous determination of trichothecene mycotoxins and zearalenone in cereals by gas chromatography-mass spectrometry. J Chromatogr A 882:23–28CrossRefPubMedGoogle Scholar

Copyright information

© Society for Mycotoxin Research and Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Sebastian Hickert
    • 1
    • 2
  • Johannes Gerding
    • 1
    • 2
  • Edson Ncube
    • 3
  • Florian Hübner
    • 1
  • Bradley Flett
    • 3
    • 4
  • Benedikt Cramer
    • 1
  • Hans-Ulrich Humpf
    • 1
    • 2
  1. 1.Institute of Food ChemistryUniversity of MünsterMünsterGermany
  2. 2.NRW Graduate School of ChemistryMünsterGermany
  3. 3.ARC-Grain Crops InstitutePotchefstroomSouth Africa
  4. 4.Unit of Environmental Sciences and ManagementNorth-West UniversityPotchefstroomSouth Africa

Personalised recommendations