aflatoxin-g2 and zearalenol

In this study, the exposure of a German population (n = 101) to mycotoxins was assessed using an LC-MS/MS urinary multibiomarker approach. Food consumption of the participants was documented with a food frequency questionnaire to correlate mycotoxin exposure with individual nutritional habits.. The presence of 23 urinary biomarkers including trichothecenes (deoxynivalenol (DON), DON-3-glucuronide (DON-3-GlcA), T-2 toxin, HT-2 toxin (HT-2, HT-2-toxin-4-glucuronide (HT-2-GlcA), fumonisins (fumonisin B1, fumonisin B2), aflatoxins (aflatoxin B1, aflatoxin G2, aflatoxin B2, aflatoxin M1), zearalenone and derivatives (zearalanone, α-zearalenol, β-zearalenol, zearalenone-14-O-glucuronide, zearalanone-14-O-glucuronide, α-zearalenol-14-O-glucuronide/β-zearalenol-14-O-glucuronide), ochratoxin A, ochratoxin alpha, enniatin B and dihydrocitrinone was evaluated using a validated, sensitive "dilute and shoot"-LC-MS/MS method applying Scheduled MRM(TM) technology. Six mycotoxins and urinary metabolites were detected (DON, DON-3-GlcA, zearalenone-14-O-glucuronide, T-2 toxin, enniatin B, and dihydrocitrinone) in 87% of the samples in single- or co-occurence. Only DON and DON-3-GlcA were detectable in quantifiable amounts. A provisional mean daily intake of 0.52 μg DON/kg body weight was calculated. No statistical evidence for the correlation of staple food intake and urinary biomarker concentration could be determined.. The results of this study suggest a low everyday exposure of the investigated German population to mycotoxins, but reveal peak exposures above the widely accepted tolerable daily intake to DON in parts of the population.

Topics: Adult; Aflatoxins; Chromatography, High Pressure Liquid; Chromatography, Liquid; Feeding Behavior; Female; Food Contamination; Food Microbiology; Fumonisins; Germany; Glucuronides; Humans; Male; Mycotoxins; Ochratoxins; Reproducibility of Results; Surveys and Questionnaires; T-2 Toxin; Tandem Mass Spectrometry; Trichothecenes; Young Adult; Zearalenone; Zeranol

A new immunoblot approach called ELISAGRAM was devised that combines the sensitivity and selectivity of competitive ELISA with the capacity of high performance thin layer chromatography (HPTLC) to separate structurally related haptens. The procedure involved (1) separation of haptens by monoclonal antibody, (3) incubation of NC with hapten-enzyme conjugate to identify unreacted antibody binding sites. (4) detection of bound enzyme conjugate with a precipitating substrate and (5) visual or densitometric assessment of inhibition bands indicative of a cross-reacting hapten. The technique was applied to two major mycotoxin families, the zearalenones and aflatoxins (AFs), which are to toxicological significance. Detection limit for zearalenone and alpha-zearalenol in the method was 300 pg/assay. AFB1, AFB2, and AFG1 were detectable at 380 pg/assay and AFG2 was detectable at 1500 pg/assay. Multiple standard curves for the zearalenones and AFs could be constructed using scanning densitometry. Cross-reactivity in ELISAGRAM curves was analogous to that found in competitive ELISA. This procedure should be widely applicable to the simultaneous quantitation and confirmation of multiple haptens with a single cross-reactive antibody.

Topics: Aflatoxin B1; Aflatoxins; Chromatography, Thin Layer; Cross Reactions; Densitometry; Enzyme-Linked Immunosorbent Assay; Haptens; Immunoblotting; Zearalenone; Zeranol