Comparison of Monoclonal Antibody-Based Enzyme-Linked Immunosorbent Assay with Thin-Layer Chromatography and Liquid Chromatography for Aflatoxin B1 Determination in Naturally Contaminated Corn and Mixed Feed

Abstract

A 1-step enzyme-linked Immunosorbent assay (EUSA) method, using a highly sensitive and specific monoclonal antibody to aflatoxin B1 (AFB1), was evaluated by comparison with other methods, Including liquid chromatography (LC) and thinlayer chromatography. The detection limit of the ELISA was as low as 100 pg/assay. AFB1 contents of naturally contaminated corn samples were determined by the 3 methods. The relationships among the methods were Investigated, and good correlations were observed. Mixed feeds were also subjected to AFB1 determination by the 3 methods. For our ELISA system, 3 types of sample preparations were tested. For analysis of mixed feed by ELISA, samples must first be purified by column chromatography. When the relationship between LC and ELISA was also investigated, results were found to have a good correlation coefficient.

Comparison of cultural and analytical methods for determination of aflatoxin production by Mississippi DeltaAspergillusisolates

“…Successful application of fluorescence enhancers in quantitative analysis has so far been limited to β-CDs, which are thought to act by complexing fluorescent mycotoxins in a hydrophobic hole in the center of the saccharide ring structure (Vazquez et al 1992). CD fluorescence enhancers have been extensively used in quantitative liquid chromatography methods for aflatoxins (Vazquez et al 1991;Hongyo et al 1992;Franco et al 1998;Chiavaro et al 2001), as well as for two other mycotoxins, ochratoxins and zearalenones (Seidel et al 1993).…”

Comparative Study of Three Different Methods for the Determination of Aflatoxins in Tahini

“…Hongyo et al (20) suggested using the same extraction solvent to increase the correlation between ELISA and HPLC methods. In addition, the same extraction procedure allows measuring only the effect of detection variances (37).…”

“…The immunological methods that have been used since the 1990s are accepted as official methods for aflatoxin determination in some food commodities; however, it is still necessary to evaluate their efficiency in different foodstuffs and to correlate their results with other accepted analytical methods (19). High-performance liquid chromatog-raphy (HPLC) and enzyme-linked immunosorbent assay (ELISA) methods were found to be highly correlated for the analysis of total aflatoxins in foodstuffs such as peanut, peanut butter, and corn and for aflatoxin B 1 in corn and mixed feeds, whereas in other foodstuffs, such as cereals, low correlations were observed (20). ELISA methods could be used only for screening purposes with the detection limit above the regulatory limits because they are not reliable when used as a quantitative method (21,22).…”

“…Similarly, the high regression obtained for HPLC and fluorometry methods may be due to the same cleanup procedure performed in both methods. Hongyo et al(20) associated the high correlation coefficient values obtained for ELISA methods to the high specificity and reproducibility of the monoclonal antibody used in their study. The type of food matrix may also have an effect on correlation values between methods.…”

Production of ultrasensitive antibodies against aflatoxin B1

“…Due to the ubiquitous nature of these fungal species and the constant possibility of their growth and aflatoxin production in various foods and feeds, there has been a concerted effort to develop sensitive aflatoxin assays. Although liquid chromatographic methods are sensitive and accurate, they are expensive, time‐consuming and unsuitable for the analysis of many samples (Hongyo et al . 1992).…”