Occurrence of relevant mycotoxins in food commodities consumed in Chile.

The aims of this study were to analyse the occurrence of aflatoxins, aflatoxin M1 (AFM1), fumonisins, ochratoxin A (OTA), patulin (PAT), zearalenone (ZEN) and deoxynivalenol (DON) in foodstuffs consumed in Chile between 2008 and 2017 and to estimate the contribution of main contaminated foodstuff in human exposure by the probable daily intake (PDI) estimation. In 9 years of surveillance, 2020 food samples were analysed with an occurrence of 18.2% and with 2.7% of the samples being over the Chilean regulation. The occurrence of mycotoxins in food were 16% for aflatoxins, 6% for AFM1, 30% for OTA, 12% for DON, 7% for PAT, 21% for fumonisins and 2% for ZEN. The estimated median PDI of DON because of bread consumption was 129.2 ng/kg bw/day for children and 96.0 ng/kg bw/day in adults. Median PDI because of capsicum consumption was 0.006 ng/kg bw/day for OTA and 0.005 ng/kg bw/day for aflatoxins. Median PDI of aflatoxins was estimated at 0.02 ng/kg bw/day for spices and 0.04 ng/kg bw/day for nuts consumption. In children, the median PDI of AFM1 for dairy consumption was 0.07 ng/kg bw/day. The derived margin of exposure (MoE) values ranged from 1133 to 8500 suggested that aflatoxins would be of public health concern. The PDI of the other mycotoxins did not show a health risk. This is the first survey of mycotoxins in food made in Chile; further research is needed to improve surveillance and guidelines based on national risk assessments and considering sensitive population groups.

Neuro-2a cell-based assay for toxicity equivalency factor - proposal and evaluation in Chilean contaminated shellfish samples.

There are two official PSP detection methods (mouse bioassay and HLPC-FLD) and a number of alternative methods. Ethical considerations have led to regulations being adopted in some countries that limit or prohibit the application of mouse bioassay. Analytical methodologies (e.g. HPLC-FLD or LC-MSMS) have the disadvantages of not being able to detect new toxins or analogues or reflecting the overall toxicity of the sample. In addition, they require highly trained personnel and expensive equipment, which are not always available. In this work, we have evaluated a method based on the Neuro-2a cell-based assay to detect substances that inhibit voltage-dependent sodium channels (Manger's method). We tested PSP standards and natural samples contaminated with PSP. Here we demonstrate that the adapted Manger's method is suitable for calculating Toxicity Equivalency Factors (TEF) for STX-analogues. The method was shown to be useful for screening contaminated natural samples in concentrations above the regulatory limit for these toxins (80 µg STX equivalents/100 g shellfish). We were able to detect PSP in 19 natural mollusc samples from South Chile despite the presence of other marine toxins. These preliminary results suggest that the method could be used as a first step in screening programmes.

Matrix effects on a cell-based assay used for the detection of paralytic shellfish toxins in bivalve shellfish samples.

Detecting marine biotoxins such as paralytic shellfish toxins (PSTs) is essential to ensuring the safety of seafood. The mouse bioassay is the internationally accepted method for monitoring PSTs, but technical and ethical issues have led to a search for new detection methods. The mouse neuroblastoma cell-based assay (Neuro-2a CBA) using ouabain and veratridine (O/V) has proven useful for the detection of PSTs. However, CBAs are sensitive to shellfish-associated matrix interferences. As the extraction method highly influences matrix interferences, this study compared three extraction protocols: Association of Official Analytical Chemists (AOAC) 2005.06, AOAC 2011.02 and an alternative liquid-liquid method. These methods were used to assess the matrix effect of extracts from four commercially important bivalve species (Chilean mussel, Magellan mussel, clam and Pacific oyster) in Neuro-2a CBA. Extracts from all three protocols caused a toxic effect in Neuro-2a cells (without O/V) when tested at a concentration of 25 mg of tissue-equivalent (TE) ml(-1). The greatest toxicity was obtained through the AOAC 2011.02 protocol, especially for the Chilean mussel and Pacific oyster extracts. Similar toxicity levels (less than 15%) were observed in all extracts at 3.1 mg TE ml(-1). When assessed in Neuro-2a CBA, AOAC 2005.06 extracts presented the lowest matrix interferences, while the highest interferences were observed for AOAC 2011.02 in Magellan mussel and clam extracts. Finally, the AOAC 2005.06 and alternative protocols were compared using Chilean mussel samples fortified with 40 and 80 µg STX per 100 g meat. The AOAC 2005.06 method demonstrated better results. In conclusion, the AOAC 2005.06 extracts exhibited the fewest interferences in the Neuro-2a CBA. Therefore, this extraction method should be considered for the implementation of Neuro-2a CBA as a high-throughput screening methodology for PST detection.

Detección del alucinógeno 25C-NBOMe en orina por dos técnicas cromatográficas / Detection of hallucinogen 25C-NBOMe in urine by two chromatographic techniques / Detecção de alucinógeno 25C-NBOME na urina através de duas técnicas cromatográficas

El consumo de drogas de síntesis ha ido en aumento. Estos nuevos derivados sintéticos son análogos estructurales de la feniletilamina N-sustituida. Este grupo ha provocado severos casos de intoxicación e incluso probablemente la muerte de varios consumidores. El principal derivado es conocido como 25C-NBOMe y se consume en estampillas idénticas al LSD. En este trabajo se desarrolla una metodología analítica para la determinación de 25C-NBOMe mediante cromatografía planar instrumental (cromatografía en capa delgada de alta resolución) y cromatografía de gases con detector de masas (CG/EM) como técnicas alternativas de fácil manejo y costo. Estas metodologías demostraron ser robustas y confiables para el propósito previsto.

Consumption of synthetic drugs has increased. These new synthetic derivatives are structural analogs of N-substituted phenylethylamine, and this group has caused severe cases of poisoning and even probably the death of several users. The main derivative is known as 25C-NBOMe and it is consumed in blotters in the same manner as LSD. In this work, an analytical methodology for 25C-NBOMe determination by instrumental planar chromatography high-performance thin-layer chromatography (HPTLC) and gas chromatography with mass detector (GC/MS) were developed as alternative techniques; they are easy to use and low cost. These methods proved to be robust and reliable for the intended purpose.

O consumo de drogas sintéticas vem aumentando. Esses novos derivados sintéticos são análogos estruturais de feniletilamina N-substituída, este grupo tem causado casos graves de intoxicação e, até mesmo, provavelmente, a morte de vários consumidores. O principal derivado é conhecido como 25C-NBOMe e consumido em selos idênticos ao LSD. Neste trabalho é desenvolvida uma metodologia analítica para a determinação de 25C-NBOMe através de cromatografia planar instrumental (cromatografia em camada delgada de alta resolução) e cromatografia gasosa com detector de massas (CG/EM) como técnicas alternativas de fácil utilização e custo. Estas metodologias demonstraram serem robustas e fiáveis para a finalidade a que se destinam.