Aflatoxin immunoaffinity columns

Sharman, M. and Gilbert, J., Automated aflatoxin analysis of foods and animal feeds using immunoaffinity column clean-up and high-performance liquid chromatographic determination, /. Chromatogr., 543, 220, 1991. 

A Patey, M Sharman, J Gilbert. Liquid chromatographic determination of aflatoxins level in peanut butter using an immunoaffinity columns clean-up. method international collaborative trials. J Ass Off Anal Chem 74(1) 76-81, 1991. 

Aflatoxins in Com, Raw Peanuts and Peanut Butter Immunoaffinity Column Method (Aflatest P column) Vicam 128... 

Another area where IAP is found to be beneficial is in the testing of afiatoxins. The development of Aflatest using IAP enrichment has allowed sample testing in minutes with limited laboratory facilities and without need for hazardous aflatoxin standards (135). An AOAC/IUPAC collaborative study was conducted to evaluate the effectiveness of the immunoaffinity column (Aflatest) for determination of aflatoxin. The method has been given official first action by the AOAC (128). 

Work Item H determination of aflatoxin Bi and total aflatoxins by immunoaffinity column clean-up and HPLC in fig paste, pistachios, peanut butter and paprika powder. 

DRAGACCI s and FREMY J M (1996), Application of immunoaffinity column cleanup to aflatoxin Ml , J Food Protect, 59, 1011-1013. 

Recently we completed an evaluation of the effectiveness of the Aflatest (Vicam) immunoaffinity column for analysis of corn, peanuts and peanut butter for aflatoxin (11). In this procedure the sample is extracted with Me0H/H20, filtered, and the extract diluted to <30% MeOH with water. An aliquot is then applied to the immunoaffinity column. The column is then washed with water and the aflatoxins eluted with methanol. Total aflatoxins are then determined by solution fluorometry with bromine (SFB), or individual toxins by reverse phase high performance liquid chromatography with post column I2 derivatization (PCD). In the collaborative study corn samples naturally contaminated with aflatoxins and samples of corn, peanuts and peanut butter spiked at 30, 20 and 10 ng/g, were analyzed by 24 collaborators. The results of this study are tabulated in Table V. 

Aflatest. This immunoaffinity column also met the lower detection requirement for the USFDA at a permissible limit of 20 ppb for aflatoxin. Qualitative results can be obtained using a florisil tip, however, the florisil tip was found difficult to interpret. Evaluation showed the quantitative analysis to be as accurate as other HPLC procedures, but much more rapid. Using the Aflatest for sample extract purification, and HPLC with post column derivatization using iodine, a limit of quantitation of 2.3 ppb for B and G is easily achieved. Sensitivity to B2 and G- is lower due to less specificity of the Aflatest antibodies to tnese toxins. 

AOAC International (2000a) Aflatoxin B1 and total aflatoxins in peanut butter, pistachio paste, tig paste, and paprika powder immunoaffinity column-liquid chromatography with postcolumn derivatization. In Official methods of analysis of AOAC International, 17th ed. Gaithersburg, MD, USA, AOAC International (AOAC Method 999.07). 

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. 

The usefulness of immunochemical techniques for the fluorimetric determination of mycotoxins has been widely demonstrated. In addition to the use of immunoaffinity column cleanup followed by LC with fluorescent detection, several FIAs have been described for the determination of aflatoxins, deoxynivalenol, and fumonisins using fluorescence polarization immunoassay or time-resolved fluor-oimmunoassay. Also, although the application of flu-oroimmunosensors to food analysis has been relatively limited, there are some examples of the use of this approach to the determination of some mycotoxins such as aflatoxin Bi and fumonisin B2. 

Aflatoxins in corn, raw peanuts, and peanut butter - immunoaffinity column (Aflatest) method... 

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... 

AOAC 2003.02, determination of aflatoxins in animal feed using immunoaffinity column cleanup and HPLC with postcolumn derivatization... 

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... 

Sharma, M. and Marquez, C. 2001. Determination of aflatoxins in domestic pet foods (dog and cat) using immunoaffinity column and HPLC. Anim. Feed Sci. Technol. 93 109-114. 

Stroka, J., von Host, C., Anklam E., and Reutter, M. 2003. Immunoaffinity column cleanup with liquid chromatography using post-column bromination for determination of aflatoxin Bl in cattle feed Collaborative study. J. AOAC Int. 86 1179-1186. 

Trucksess, M. W., Stack, M. E., Nesheim, S., Page, S. W., Albert, R., Hansen, T. J., and Donahue, K. F. 1991. Immunoaffinity column coupled with solution fluorometry or liquid chromatography postcolumn derivatization for determination of aflatoxins in corn, peanuts and peanut butter Collaborative study. J. Assoc. Offi. Anal. Chem. 74 81-88. 

Dragacci, S., Grosso, R, and Gilbert, J. 2001. Immunoaffinity column cleanup with liquid chromatography for determination of aflatoxin Ml in liquid milk Collaborative study. J. AOACInt. 84, 2 437 43. 

Muscarella, M., Lo Magro, S., Palermo, C., and Centonze, D. 2007. Validation according to European Commission Decision 2002/657/EC of a confirmatory method for aflatoxin Ml in milk based on immunoaffinity columns and high performance liquid chromatography with fluorescence detection. Anal. Chim. Acta 594 257-264. 

Ip, S. P. and Che, C. T. 2006. Determination of aflatoxins in Chinese medicinal herbs by high-performance liquid chromatography using immunoaffinity column cleanup Improvement of recovery. / Chromatogr.A 1135 241-244. 

Basically the cleanup step consists of one of the two approaches already described in the aflatoxin section (a) the use of solid-phase extraction (SPE) columns, or (b) the use of immunoaffinity (IA) columns. 

Immunoaffinity (lA) methods are based on the principle of molecular recognition via very selective antigen-antibody interactions. Their use for sample preparation prior to TLC has been mostly for determination of toxins. The following are selected examples the steroid animal drug trenbolone and its metabolite in bovine urine " fumonisin B1 in corn with methanol-water (80 20) extraction followed by lA column cleanup " aflatoxins Bl, B2, Gl, and G2 in foods regulated within... 

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