Aflatoxin M1 ELISA Kit for Cheese Sample

Medicinal plants are often consumed as infusions with boiled water. Scarce information is available in the literature about the migration of mycotoxins into the resulting beverage and/or the effects of the infusion procedure on the final mycotoxin contents. The aim of the present study was to investigate the impact of the infusion process on mycotoxin contents during medicinal plant preparation. For this purpose, the contents of aflatoxins (AFs) [aflatoxin B1 (AFB1), aflatoxin B2 (AFB2), aflatoxin G1 (AFG1), aflatoxin G2 (AFG2)], zearalenone (ZEA), enniatins (ENNs) [enniatin B (ENNB), enniatin B1 (ENNB1), enniatin A (ENNA), enniatin A1 (ENNA1)] and beauvericin (BEA) were analyzed in 224 samples of medicinal plants and in their resulting beverages. The quick, easy, cheap, effective, rugged and safe extraction method (QuEChERS) was applied to the medicinal plants while the dispersive liquid-liquid microextraction procedure (DLLME) was applied to their infusions, and the mycotoxins were determined by liquid chromatography coupled to ion trap tandem mass spectrometry (LC-MS/MS-IT). The results revealed that ZEA, ENNB, ENNB1, AFB2, AFG1 and AFG2 were detected in the beverages with incidences of <= 6% and at concentrations from less than the limit of quantification (LOQ) to 82.2 mu g/L. Mycotoxins reduction ranged from 74 to 100% after the infusion process. The risk assessment revealed that the estimated daily intakes (EDIs) obtained for ZEA, ENNB and ENNB1 were far below the tolerable daily intakes (TDIs) established.

In vitro assessment of genotoxicity as an early warning tool for carcinogenicity mainly relies on recording cytogenetic damages (micronuclei, nucleoplasmic bridges) in tumour-derived mammalian cell lines like V79 or CHO. The forecasting power of the corresponding standardised test is based on epidemiological evidence between micronuclei frequencies and cancer incidence. As an alternative to destructive staining of nuclear structures a fish stem cell line transgenic for a fusion protein of histone 2B (H2B) and enhanced green fluorescent protein (eGFP) was established. The cells are derived from koi carp brain (KCB) and distinguish from mammalian culturable cells by non-tumour-driven self-renewal. This technology enables the analysis of genotoxic- and malign downstream effects in situ in a combined approach. In proof-of concept-experiments, we used known carcinogens (4-Nitroquinoline 1-oxide, colchicine, diethylstilbestrol, ethyl methanesulfonate) and observed a significant increase in micronuclei (MNi) frequencies in a dose-dependent manner. The concentration ranges for MNi induction were comparable to human/mammalian cells (i.e. VH-16, CHL and HepG2). Cannabidiol caused the same specific cytogenetic damage pattern as observed in human cells, in particular nucleoplasmic bridges. Metabolic activation of aflatoxin B1 and cyclophosphamide could be demonstrated by pre-incubation of the test compounds using either conventional rat derived S9 mix as well as an in vitro generated biotechnological alternative product ewoS9R. The presented high throughput live H2B-eGFP imaging technology using non-transformed stem cells opens new perspectives in the field of in vitro toxicology. The technology offers experimental access to investigate the effects of carcinogens on cell cycle control, gene expression pattern and motility in the course of malign transformation. The new technology enables the definition of Adverse Outcome Pathways leading to malign cell transformation and contributes to the replacement of animal testing. Summary: Complementation of genotoxicity testing by addressing initiating events leading to malign transformation is suggested. A vertebrate cell model showing "healthy" stemness is recommended, in contrast to malign transformed cells used in toxicology/oncocology.