Ochratoxin immunoaffinity columns

B Zimmerli, R Dick. 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 Chrom B 666 85-99,1995. 

M Sharman, S Mac Donald, J Gilbert. Automated liquid chromatographic determination of ochratoxin A in cereals and animal products using immunoaffinity column clean-up. J Chiom 603 285-289, 1992. 

A Pittet, D Tornare, A Huggett, R Viani. Liquid chromatographic determination of ochratoxin A in pure and adulterated soluble coffee using an immunoaffinity column cleanup procedure. J Agric Food Chem 44 3564-3569,1996. 

International Organization of Vine and Wine, Measuring ochratoxin A in wine after going through an immunoaffinity column and HPLC with fluorescence detection, Compendium of International Methods of Wine and Must Analysis Vol. 2 (2006). 

Visconti, A., Pascale, M. and Centonze, G. (1999) Determination of ochratoxin A in wine by means of immunoaffinity column clean-up and high-performance liquid chromatography,/. Chromatogr. A, 864 (1), 89-101. 

Chan, D., MacDonald, S.J., Boughtflower, V. Brereton, P. (2004) Simultaneous determination of aflatoxins and ochratoxin A in food using a fully automated immunoaffinity column clean-up and liquid chromatography-fluorescence detection. J. Chromatogr. A, 1059, 13-16. 

Methods for analysing ochratoxin A were thoroughly reviewed in the previous report (Annex 1, reference 153). At that time, it was reported that validated analytical methods were available for the determination of ochratoxin A in maize, barley, rye, wheat, wheat bran, wholemeal wheat, roasted coffee, wine and beer. The best methods used LC following cleanup using immunoaffinity columns. Over 40 methods were reviewed. This review is limited to recent developments. 

For many solid samples, a solvent extraction step is still necessary. Extraction with acidic chloroform, acidic ethyl acetate containing 2 mol sodium chloride/l or methanol-sodium carbonate (1%), followed by back-extraction to sodium bicarbonate, has been recommended (Monaci Palmisano, 2004). After washing the column, ochratoxin A can be eluted with methanol and, if necessary, further purified through an immunoaffinity column. 

T. (2001) Determination of ochratoxin A in baby food by immunoaffinity column cleanup with liquid chromatography interlaboratory study. J. AOAC Int 84,1445-1452. 

Entwisle, A.C., Williams, A.C., Mann, P.J., Slack, P.T. Gilbert, J. (2000) Liquid chromatographic method with immunoaffinity column cleanup for determination of ochratoxin A in barley collaborative study. J. AOAC Int. 83,1377-1383. 

Pardo, E., Marin, S., Ramos, A.J. Sanchis, V. (2004) Occurrence of ochratoxigenic fungi and ochratoxin A in green coffee from different origins. Food Sci. Technol. Int. 10,45-49. Pascale, M. Visconti, A. (2001) Rapid method for the detennination of ochratoxin A in urine by immunoaffinity column clean-up and high-perfonnance liquid chromatography. 

AOAC 2000.02, determination of patulin in clear and cloudy apple juices and apple puree using HPLC AOAC 2000.03, determination of ochratoxin in barley using HPLC and immunoaffinity cleanup AOAC 2000.08, aflatoxin Ml in liquid milk using immunoaffinity column cleanup and HPLC AOAC 2000.09, determination of ochratoxin in a roasted coffee using HPLC AOAC 2000.16, aflatoxin Bl in baby food using an immunoaffinity column and HPLC AOAC 2001.01, determination of ochratoxin A in wine using immunoaffinity column cleanup and HPLC... 

Aflatoxins Deoxynivalenol Fumonisins Monilifomin Ochratoxin Zearalenone TLC, HPLC, ELISA, immunoaffinity column GC, HPLC, ELISA, immunoaffinity column HPLC, ELISA, immunoaffinity column HPLC TLC, HPLC, ELISA, immunoaffinity column TLC, HPLC, ELISA, immunoaffinity coluum... 

Santos, E.A. Vargas, E.A. Immunoaffinity column cleanup and thin layer chromatography for determination of ochratoxin A in green coffee. Food Addit. Contam. 2002,19 (5), 447-458. 

The main disadvantages of an immunoaffinity cleanup for OA could in some cases be the lack of specificity for ochratoxin A (since a cross-reaction with ochratoxin C can occur (60)), the potential contamination with release of ochratoxin A from the support of the IA (61), and the high cost of each column, although some authors considered the possibility of regenerating the column for further analyses (30). 

---