LC-MS/MS multi-analyte method for mycotoxin determination in food supplements

A multi-analyte method for the liquid chromatography-tandem mass spectrometric determination of mycotoxins in food supplements is presented. The analytes included A and B trichothecenes (nivalenol, deoxynivalenol, 3-acetyldeoxynivalenol, 15-acetyldeoxynivalenol, neosolaniol, fusarenon-X, diacetoxyscirpenol, HT-2 toxin and T-2 toxin), aflatoxins (aflatoxin-B1, aflatoxin-B2, aflatoxin-G1 and aflatoxin-G2), Alternaria toxins (alternariol, alternariol methyl ether and altenuene), fumonisins (fumonisin-B1, fumonisin-B2 and fumonisin-B3), ochratoxin A, zearalenone, beauvericin and sterigmatocystin. Optimization of the simultaneous extraction of these toxins and the sample pretreatment procedure, as well as method validation were performed on maca (Lepidium meyenii) food supplements. The results indicated that the solvent mixture ethyl acetate/formic acid (95:5, v/v) was the best compromise for the extraction of the analytes from food supplements. Liquid–liquid partition with n-hexane was applied as partial clean-up step to remove excess of co-extracted non-polar components. Further clean-up was performed on Oasis HLB™ cartridges. Samples were analysed using an Acquity UPLC system coupled to a Micromass Quattro Micro triple quadrupole mass spectrometer equipped with an electrospray interface operated in the positive-ion mode. Limits of detection and quantification were in the range of 0.3–30 ng g−1 and 1–100 ng g−1, respectively. Recovery yields were above 60% for most of the analytes, except for nivalenol, sterigmatocystine and the fumonisins. The method showed good precision and trueness. Analysis of different food supplements such as soy (Glycine max) isoflavones, St John's wort ( Hypericum perforatum), garlic (Allium sativum), Ginkgo biloba, and black radish (Raphanus niger) demonstrated the general applicability of the method. Due to different matrix effects observed in different food supplement samples, the standard addition approach was applied to perform correct quantitative analysis. In 56 out of 62 samples analysed, none of the 23 mycotoxins investigated was detected. Positive samples contained at least one of the toxins fumonisin-B1, fumonisin-B2, fumonisin-B3 and ochratoxin A.

[1]  P. Scott,et al.  Mycotoxins in botanicals and dried fruits: A review , 2008, Food additives & contaminants. Part A, Chemistry, analysis, control, exposure & risk assessment.

[2]  R. Krska,et al.  Development and validation of a liquid chromatography/tandem mass spectrometric method for the determination of 39 mycotoxins in wheat and maize. , 2006, Rapid communications in mass spectrometry : RCM.

[3]  L. van der Merwe,et al.  Mycotoxin contamination of dietary and medicinal wild plants in the Eastern Cape Province of South Africa. , 2006, Journal of agricultural and food chemistry.

[4]  D. Heber,et al.  An analysis of nine proprietary Chinese red yeast rice dietary supplements: implications of variability in chemical profile and contents. , 2001, Journal of alternative and complementary medicine.

[5]  E. Razzazi-Fazeli,et al.  Contamination of Aflatoxins in Herbal Medicinal Products in Thailand , 2004, Mycopathologia.

[6]  H. S. Hussein,et al.  Toxicity, metabolism, and impact of mycotoxins on humans and animals. , 2001, Toxicology.

[7]  Gopal Reddy,et al.  Occurrence of ochratoxin A in black pepper, coriander, ginger and turmeric in India , 2001, Food additives and contaminants.

[8]  G. Ware,et al.  Preparative Method for Isolating α-Zearalenol and Zearalenone Using Extracting Disk , 1999 .

[9]  P. G. Thiel,et al.  Worldwide survey of fumonisin contamination of corn and corn-based products. , 1996, Journal of AOAC International.

[10]  L. K. Sørensen,et al.  Determination of mycotoxins in bovine milk by liquid chromatography tandem mass spectrometry. , 2005, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[11]  Y. Ueno,et al.  Simultaneous determination of trichothecene mycotoxins and zearalenone in cereals by gas chromatography-mass spectrometry. , 2000, Journal of chromatography. A.

[12]  V. Betina.  Chapter 7 Thin-Layer Chromatography of Mycotoxins , 1993 .

[13]  A. Wilkins,et al.  Determination of Penicillium mycotoxins in foods and feeds using liquid chromatography-mass spectrometry. , 2002, Journal of chromatography. A.

[14]  M. Halt,et al.  Moulds and mycotoxins in herb tea and medicinal plants , 1998, European Journal of Epidemiology.

[15]  H Valenta,et al.  Chromatographic methods for the determination of ochratoxin A in animal and human tissues and fluids. , 1998, Journal of chromatography. A.

[16]  W. Marasas,et al.  Determination of the Fusarium mycotoxins, fusaproliferin and beauvericin by high-performance liquid chromatography-electrospray ionization mass spectrometry. , 1999, Journal of chromatography. A.

[17]  Carol Weaver,et al.  Aflatoxins in ginseng roots , 2006, Food additives and contaminants.

[18]  Michael Rychlik,et al.  Stable isotope dilution assays in mycotoxin analysis , 2008, Analytical and bioanalytical chemistry.

[19]  C. Cavaliere,et al.  Determination of type B trichothecenes and macrocyclic lactone mycotoxins in field contaminated maize , 2005 .

[20]  T. Rundberget,et al.  Instrumental methods for determination of nonmacrocyclic trichothecenes in cereals, foodstuffs and cultures , 1998 .

[21]  M. Haidukowski,et al.  Assessment of toxigenic fungi on Argentinean medicinal herbs. , 2004, Microbiological research.

[22]  G. Shephard.  Chromatographic determination of the fumonisin mycotoxins. , 1998, Journal of chromatography. A.

[23]  M. Spanjer,et al.  LC–MS/MS multi-method for mycotoxins after single extraction, with validation data for peanut, pistachio, wheat, maize, cornflakes, raisins and figs , 2008, Food additives & contaminants. Part A, Chemistry, analysis, control, exposure & risk assessment.

[24]  R. Krska,et al.  The state-of-the-art in the analysis of estrogenic mycotoxins in cereals , 2001, Fresenius' journal of analytical chemistry.

[25]  K. Schwadorf,et al.  Determination of α- and β-zearalenol and zearalenone in cereals by gas chromatography with ion-trap detection , 1992 .

[26]  C. Hummert,et al.  Analysis of Fusarium Toxins Using LC-MS-MS: Application to Various Food and Feed Matrices , 2004 .

[27]  U. Thrane,et al.  Fast methods for screening of trichothecenes in fungal cultures using gas chromatography-tandem mass spectrometry. , 2001, Journal of chromatography. A.

[28]  M. Nakajima,et al.  Evaluation of a method to determine the natural occurrence of aflatoxins in commercial traditional herbal medicines from Malaysia and Indonesia. , 2005, Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association.

[29]  C. V. Van Peteghem,et al.  Development of a liquid chromatography/tandem mass spectrometry method for the simultaneous determination of 16 mycotoxins on cellulose filters and in fungal cultures. , 2006, Rapid communications in mass spectrometry : RCM.

[30]  G. Ware,et al.  Preparative method for isolating alpha-zearalenol and zearalenone using extracting disk. , 1999, Journal of AOAC International.

[31]  G. Omurtag,et al.  Determination of fumonisins B1 and B2 in herbal tea and medicinal plants in Turkey by high-performance liquid chromatography. , 2004, Journal of food protection.

[32]  George A. Bean,et al.  Toxigenic Aspergillus flavus and aflatoxins in Sri Lankan medicinal plant material , 1991, Mycopathologia.

[33]  A. Logrieco,et al.  Simultaneous determination of beauvericin, enniatins, and fusaproliferin by high performance liquid chromatography. , 2000, Journal of agricultural and food chemistry.

[34]  Sarah De Saeger,et al.  Development of a multi-mycotoxin liquid chromatography/tandem mass spectrometry method for sweet pepper analysis. , 2009, Rapid communications in mass spectrometry : RCM.

[35]  R. Krska,et al.  The state-of-the-art in the analysis of type-A and -B trichothecene mycotoxins in cereals , 2001, Fresenius' journal of analytical chemistry.

[36]  D M Goldberg,et al.  Assay of ochratoxin A in wine and beer by high-pressure liquid chromatography photodiode array and gas chromatography mass selective detection. , 2001, Journal of agricultural and food chemistry.

[37]  Hamed K. Abbas,et al.  Aflatoxin and fumonisin contamination of corn (maize, Zea mays) hybrids in Arkansas , 2006 .

[38]  J. Fink-Gremmels.  Mycotoxins: their implications for human and animal health. , 1999, The Veterinary quarterly.

[39]  J. Young,et al.  Detection and characterization of fumonisin mycotoxins as their methyl esters by liquid chromatography/particle‐beam mass spectrometry , 1993 .

[40]  N. Camper,et al.  Mycotoxins in Root Extracts of American and Asian Ginseng Bind Estrogen Receptors α and β , 2004 .

[41]  H. M. Martins,et al.  Fumonisins B1 and B2 in black tea and medicinal plants. , 2001, Journal of food protection.

[42]  V. Betina,et al.  Chromatography of mycotoxins : techniques and applications , 1993 .

[43]  H. Humpf,et al.  Quantitative analysis of Fusarium mycotoxins in maize using accelerated solvent extraction before liquid chromatography/atmospheric pressure chemical ionization tandem mass spectrometry , 2004, Food additives and contaminants.

[44]  Rudolf Krska,et al.  Mycotoxin analysis: An update , 2008, Food additives & contaminants. Part A, Chemistry, analysis, control, exposure & risk assessment.

[45]  R. Thakur,et al.  Analysis of fumonisin B1 by negative‐ion thermospray mass spectrometry , 1994 .

[46]  R. I,et al.  Mycotoxicological control on raw material and tablets of cascara sagrada (Rhamnus purshiana) , 2008, Mycotoxin Research.

[47]  J. Fink-Gremmels.  Mycotoxins in cattle feeds and carry-over to dairy milk: A review , 2008, Food additives & contaminants. Part A, Chemistry, analysis, control, exposure & risk assessment.

[48]  Joseph M Betz,et al.  Use of multitoxin immunoaffinity columns for determination of aflatoxins and ochratoxin A in ginseng and ginger. , 2007, Journal of AOAC International.

[49]  Mei-Hua Yang,et al.  Immunoaffinity Column Clean-Up and Liquid Chromatography with Post-Column Derivatization for Analysis of Aflatoxins in Traditional Chinese Medicine , 2005 .

[50]  M. Jestoi,et al.  Determination of selected mycotoxins in mould cheeses with liquid chromatography coupled to tandem with mass spectrometry , 2005, Food additives and contaminants.

[51]  R. Krska,et al.  Rapid simultaneous determination of major type A- and B-trichothecenes as well as zearalenone in maize by high performance liquid chromatography-tandem mass spectrometry. , 2005, Journal of chromatography. A.

[52]  W. Langseth.  Mycotoxin production and cytotoxicity of Fusarium strains isolated from Norwegian cereals , 1998, Mycopathologia.

[53]  P. Galtier.  Biological fate of mycotoxins in animals , 1998 .