Impact of pH on the Stability and the Cross-Reactivity of Ochratoxin A and Citrinin

Mycotoxins are secondary metabolites produced by several fungi contaminating crops. In several countries, the maximum permitted levels of mycotoxins are found in foodstuffs and feedstuffs. The common strategy of mycotoxin analysis involves extraction, clean-up and quantification by chromatography. In this paper, we analyzed the reasons of underestimation of ochratoxin A (OTA) content in wine, and overestimation of OTA in wheat, depending on the pH of the clean-up step and the simultaneous presence of citrinin (CIT). We demonstrated that the increase of pH by adding polyethylene glycol (PEG) to wine led to an underestimation of OTA by conversion of OTA into open ring ochratoxin A OP-OA. In comparing three methods of extraction and clean-up for the determination of OTA and CIT in wheat—(i) an inter-laboratory validated method for OTA in cereals using immunoaffinity column clean-up (IAC) and extraction by acetonitrile/water; (ii) a validated method using IAC and extraction with 1% bicarbonate Na; and (iii) an in-house validated method based on acid liquid/liquid extraction—we observed an overestimation of OTA after immunoaffinity clean-up when CIT is also present in the sample, whereas an underestimation was observed when OTA was alone. Under neutral and alkaline conditions, CIT was partially recognized by OTA antibodies.

[1]  Jean-Louis Marty,et al.  Label-free impedimetric immunosensor for sensitive detection of ochratoxin A. , 2009, Biosensors & bioelectronics.

[2]  P. Cunniff Official Methods of Analysis of AOAC International , 2019 .

[3]  H. Iwahashi,et al.  Evaluation of toxicity of the mycotoxin citrinin using yeast ORF DNA microarray and Oligo DNA microarray , 2007, BMC Genomics.

[4]  E. González-Peñas,et al.  Determination of ochratoxin A in wine using liquid-phase microextraction combined with liquid chromatography with fluorescence detection. , 2004, Journal of chromatography. A.

[5]  H. Raistrick,et al.  On the Production and Chemical Constitution of a New Yellow Colouring Matter, Citrinin, Produced from Glucose by Penicillium citrinum Thom , 1931 .

[6]  M. C. Elias,et al.  Efeito da umidade e do período de armazenamento hermético na contaminação natural por fungos e a produção de micotoxinas em grãos de aveia , 2006 .

[7]  Jean-Louis Marty,et al.  Novel highly-performing immunosensor-based strategy for ochratoxin A detection in wine samples. , 2008, Biosensors & bioelectronics.

[8]  S. Pepeljnjak,et al.  Cytotoxicity, genotoxicity and apoptosis induced by ochratoxin A and citrinin in porcine kidney PK15 cells: Effects of single and combined mycotoxins , 2007 .

[9]  Delia B. Rodriguez-Amaya,et al.  Aflatoxin M1 and ochratoxin A in a human milk bank in the city of São Paulo, Brazil , 2005 .

[10]  A. Dobson,et al.  Mycotoxin production by Aspergillus, Fusarium and Penicillium species. , 1998, International journal of food microbiology.

[11]  C. Lino,et al.  A review on ochratoxin A occurrence and effects of processing of cereal and cereal derived food products. , 2010, Food microbiology.

[12]  R. Manderville,et al.  Ochratoxin A: An overview on toxicity and carcinogenicity in animals and humans. , 2007, Molecular nutrition & food research.

[13]  Hans P. van Egmond,et al.  Worldwide regulations for mycotoxins. , 2002 .

[14]  C. Lino,et al.  Ochratoxin A in feed of food-producing animals: an undesirable mycotoxin with health and performance effects. , 2011, Veterinary microbiology.

[15]  J. Frisvad,et al.  Ochratoxin A producing Penicillium verrucosum isolates from cereals reveal large AFLP fingerprinting variability , 2005, Journal of applied microbiology.

[16]  F. Pont,et al.  Evidence of a new dechlorinated ochratoxin A derivative formed in opossum kidney cell cultures after pretreatment by modulators of glutathione pathways: correlation with DNA-adduct formation. , 2006, Molecular nutrition & food research.

[17]  M. Castegnaro,et al.  Advantages and drawbacks of immunoaffinity columns in analysis of mycotoxins in food. , 2006, Molecular nutrition & food research.

[18]  H. V. van Egmond Worldwide regulations for mycotoxins. , 2002, Advances in experimental medicine and biology.

[19]  Setsuko Tabata,et al.  HPLC-FLおよびLC/MS/MSによる食品中のオクラトキシンA, Bおよびシトリニンの同時分析法 , 2008 .

[20]  K. Kamata,et al.  [Simultaneous determination of ochratoxin A, B and citrinin in foods by HPLC-FL and LC/MS/MS]. , 2008, Shokuhin eiseigaku zasshi. Journal of the Food Hygienic Society of Japan.

[21]  M. Zón,et al.  Development of an amperometric biosensor based on peroxidases to quantify citrinin in rice samples. , 2013, Bioelectrochemistry.

[22]  J. Gilbert,et al.  Survey for co-occurrence of ochratoxin A and aflatoxin B1 in dried figs in Turkey by using a single laboratory-validated alkaline extraction method for ochratoxin A. , 2005, Journal of food protection.

[23]  K. Jørgensen Occurrence of ochratoxin A in commodities and processed food--a review of EU occurrence data. , 2005, Food additives and contaminants.

[24]  E. Pedersen,et al.  OCCURRENCE OF OCHRATOXIN A AND CITRININ IN CEREALS ASSOCIATED WITH MYCOTOXIC PORCINE NEPHROPATHY , 2009 .

[25]  J Gilbert,et al.  Liquid chromatographic method with immunoaffinity column cleanup for determination of ochratoxin A in barley: collaborative study. , 2000, Journal of AOAC International.

[26]  F. Palmisano,et al.  Determination of ochratoxin A in pig tissues by liquid–liquid extraction and clean-up and high-performance liquid chromatography , 2004, Analytical and bioanalytical chemistry.

[27]  S. Pepeljnjak,et al.  Co-occurrence of Aflatoxins, Ochratoxin A, Fumonisins, and Zearalenone in Cereals and Feed, Determined by Competitive Direct Enzyme-Linked Immunosorbent Assay and Thin-Layer Chromatography , 2009, Arhiv za higijenu rada i toksikologiju.

[28]  A. G. Frenich,et al.  Multi-mycotoxin analysis in eggs using a QuEChERS-based extraction procedure and ultra-high-pressure liquid chromatography coupled to triple quadrupole mass spectrometry. , 2011, Journal of chromatography. A.

[29]  Liangqia Guo,et al.  Microsphere-based flow cytometric immunoassay for the determination of citrinin in red yeast rice. , 2012, Food chemistry.

[30]  R. Manderville,et al.  Combined Toxic Effects of Ochratoxin A and Citrinin, In Vivo and In Vitro , 2008 .

[31]  Jian-hui Jiang,et al.  Ultrasensitive electrochemical immunosensor for ochratoxin A using gold colloid-mediated hapten immobilization. , 2009, Analytical biochemistry.

[32]  Julio Raba,et al.  Citrinin (CIT) determination in rice samples using a micro fluidic electrochemical immunosensor. , 2011, Talanta.

[33]  睦代 門平,et al.  Food and Agriculture Organization (FAO) , 2002, International Organizations and the Law of the Sea 2001.

[34]  R. Manderville,et al.  An update on direct genotoxicity as a molecular mechanism of ochratoxin a carcinogenicity. , 2012, Chemical research in toxicology.

[35]  W. Horwitz Official Methods of Analysis , 1980 .

[36]  A. Pfohl-Leszkowicz,et al.  Preliminary data on the presence of mycotoxins (ochratoxin A, citrinin and aflatoxin B1) in black table olives "Greek style" of Moroccan origin. , 2006, Molecular nutrition & food research.

[37]  R. Dietrich,et al.  Co-occurrence of ochratoxin A and citrinin in cereals from Bulgarian villages with a history of Balkan endemic nephropathy. , 2000, Journal of agricultural and food chemistry.

[38]  P. Burdaspal,et al.  Liquid Chromatographic Method with Immunoaffinity Column Cleanup for Determination of Ochratoxin A in Barley: Collaborative Study , 2000 .

[39]  A. Pfohl-Leszkowicz,et al.  Occurrence of aflatoxin B1, citrinin and ochratoxin A in rice in five provinces of the central region of Vietnam , 2007 .

[40]  A. Pfohl-Leszkowicz,et al.  Ochratoxin A and Aristolochic Acid Involvement in Nephropathies and Associated Urothelial Tract Tumours , 2009, Arhiv za higijenu rada i toksikologiju.

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

[42]  W. Horwitz,et al.  Official methods of analysis of AOAC International , 2010 .

[43]  Meihua Yang,et al.  Molecularly imprinted polymer-based solid phase clean-up for analysis of ochratoxin A in ginger and LC-MS/MS confirmation , 2013 .

[44]  M. Sabater-Vilar,et al.  Mutagenicity of commercial Monascus fermentation products and the role of citrinin contamination. , 1999, Mutation research.

[45]  M. Castegnaro,et al.  Analysis of some breakfast cereals on the French market for their contents of ochratoxin A, citrinin and fumonisin B1: development of a method for simultaneous extraction of ochratoxin A and citrinin , 2005 .

[46]  Ö. Tokuşoğlu,et al.  Citrinin risk in black and green table olives: simultaneous determination with ochratoxin-A by optimized extraction and IAC-HPLC-FD. , 2010 .

[47]  R. Manderville,et al.  Structure-activity relationships for the fluorescence of ochratoxin A: insight for detection of ochratoxin A metabolites. , 2008, Analytica chimica acta.

[48]  R. Manderville,et al.  Bioactivation and DNA adduction as a rationale for ochratoxin A carcinogenesis , 2008 .

[49]  Yanghao Guo,et al.  Preparation of synthetic antigen and monoclonal antibody for indirect competitive ELISA of citrinin , 2012 .

[50]  Mariana Tozlovanu,et al.  Ochratoxin A in Roasted Coffee from French Supermarkets and Transfer in Coffee Beverages: Comparison of Analysis Methods , 2010, Toxins.

[51]  A. Visconti,et al.  Determination of ochratoxin A in wine by means of immunoaffinity column clean-up and high-performance liquid chromatography. , 1999, Journal of chromatography. A.

[52]  V. Dohnal,et al.  Occurrence of ochratoxin A and citrinin in Czech cereals and comparison of two HPLC methods for ochratoxin A detection , 2010, Food additives & contaminants. Part A, Chemistry, analysis, control, exposure & risk assessment.

[53]  N. Kitabatake,et al.  A Major Decomposition Product, Citrinin H2, from Citrinin on Heating with Moisture , 2002, Bioscience, biotechnology, and biochemistry.

[54]  Dawei Deng,et al.  Surface plasmon resonance biosensor for the detection of ochratoxin A in cereals and beverages. , 2009, Analytica chimica acta.

[55]  W. K. Anslow,et al.  Studies in the biochemistry of micro-organisms , 1933 .

[56]  R. Manderville,et al.  New molecular and field evidences for the implication of mycotoxins but not aristolochic acid in human nephropathy and urinary tract tumor. , 2007, Molecular nutrition & food research.

[57]  Ingrid Bazin,et al.  Rapid Visual Tests: Fast and Reliable Detection of Ochratoxin A , 2010, Toxins.

[58]  J. Câmara,et al.  A survey of the occurrence of ochratoxin A in Madeira wines based on a modified QuEChERS extraction procedure combined with liquid chromatography–triple quadrupole tandem mass spectrometry , 2013 .

[59]  M Castegnaro,et al.  Balkan endemic nephropathy and associated urinary tract tumours: a review on aetiological causes and the potential role of mycotoxins , 2002, Food additives and contaminants.

[60]  A. Agostiano,et al.  Effect of β-cyclodextrin on spectroscopic properties of ochratoxin A in aqueous solution , 2007 .