As the majority of consumed food items were derived from cereals,

As the majority of consumed food items were derived from cereals, the percentage of see more carbohydrates in the overall diet was exceptionally

high. The time of consumption, ingested daily quantities and concentrations of major mycotoxins are reported in Table 1. In addition, the total quantity of mycotoxins ingested during a day of intervention is stated. Vegetables, fruits and drinks (predominantly water) which are usually not likely to be contaminated with mycotoxins were consumed ad libitum. Food items consumed during the intervention diet as well as during the cereal reduced diet (rice) were analyzed for their mycotoxin contamination level prior consumption. Urine samples were collected as 24 h urine throughout the study, for which on average 7.5 spot urine samples were combined. A 24 h period lasted from 7 am to 7 am on the next day to include the first morning urine in the sample of the previous day. The rationale was based on an experiment which revealed that first morning void is well suited to represent exposure of the prior day (Turner et al., 2009). In addition, an aliquot of each spot urine sample was taken starting on day three to investigate the kinetics of DON/ZEN metabolism and excretion and to investigate if sampling of first morning void is feasible. No spot samples were collected on the first two days, as they were designed to reach blank samples only. Samples were brought to the laboratory in the morning and frozen immediately

at −20 °C. Cereal based food samples (n = 23) were purchased from supermarkets in Vienna and analyzed on their mycotoxin contamination levels using the method of Sulyok et al. ( Sulyok et al., 2007). Samples with relatively high deoxynivalenol and zearalenone concentrations were chosen to create a reasonable diet plan (see Table 1 and Table 2). However, none of the samples exceeded the regulatory limits Avelestat (AZD9668) currently enforced in the European Union ( European Commission, 2006). This study was permitted by the ethics commission of the government of Lower Austria. Determination of urinary mycotoxins and metabolites was carried out using a recently developed and validated multi-biomarker method (Warth et al., 2012b).

This method does not require any sample preparation other than centrifugation and dilution and enables to directly quantify glucuronides of deoxynivalenol and zearalenone in human urine besides their parent toxins as well as ten other relevant mycotoxins or metabolites. Briefly, samples were allowed to reach room temperature, centrifuged for 3 min at 5600 × g and diluted 1:10 with dilution solvent (ACN/H2O: 10/90). Five μL of the diluted sample (corresponding to 0.5 μL urine) were injected to a 5500 Q-Trap system (AB Sciex, Foster City, CA) equipped with an Agilent 1290 UHPLC system (Waldbronn, Germany). Analytes were separated on an Atlantis T3 column (3.0 × 150 mm, Waters, Wexford, Ireland) with 3 μm particle size and a C18 pre-column. Gradient elution at 35 °C was performed within 18 min.

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