Analysis of patulin in pear- and apple-based foodstuffs by liquid chromatography electrospray ionization tandem mass spectrometry.

A liquid chromatography electrospray ionization tandem mass spectrometry method for the determination of patulin in apple- and pear-based foodstuffs was developed. The sample preparation is based on the QuEChERS procedure involving an initial extraction step with water and acetonitrile, followed by a partitioning step after the addition of magnesium sulfate and sodium chloride. The cleanup was performed by using dispersive solid-phase extraction with a mixture of magnesium sulfate, primary secondary amine sorbent, and n-octadecylsiloxane sorbent added together to the extract. The cleaned extract was finally evaporated and reconstituted in water prior to injection. Quantitation was performed by isotope dilution using ((13)C(7))-patulin as internal standard. The method was first fully validated in three different baby food products including apple-pear juice, apple-pear puree, and infant cereals. Then the scope of application of the method was extended to pear concentrate, raw apples, apple flakes (naturally contaminated), dried apples, and yogurt. The sensitivity achieved by the method in all matrices gave limits of detection (LOD) and quantitation (LOQ) of ≤0.5 and ≤10 µg/kg, respectively, which was compliant with maximum levels settled in Commission Regulation (EC) No. 1881/2006. Method performances for all matrices also fulfilled the criteria established in the CEN/TR 16059:2010 document. Indeed, recoveries were within the 94-104% range; relative standard deviations of repeatability (RSD(r)) and intermediate reproducibility (RSD(IR)) were ≤7.5 and ≤13.0%, respectively, and trueness in an infant apple drink (FAPAS 1642) was measured at 99%.

A new PCR-based bioassay strategy for the detection of type A trichothecenes in food.

Type A trichothecenes (primarily T-2 and HT-2 toxins) are common fungal metabolites found in a wide range of grains and other field crops grown in temperate climatic zones. By acting as potent inhibitors of protein synthesis, T-2 and HT-2 exert adverse effects particularly against rapidly proliferating tissues, including the bone marrow, the immune system and epithelial cells. Based on toxicity studies in laboratory and farm animals, a temporary tolerable daily intake for the sum of T-2 and HT-2 has been issued in the European Union. However, exposure assessments suggest that the combined intake of these natural compounds exceeds in many cases the proposed threshold. To further protect the consumers, it is therefore necessary to screen a large number of food samples for parts per billion levels of both T-2 and HT-2. Towards that goal, we are the first to report that these two type A trichothecenes induce fast and high-amplitude transcriptional changes in cultured human breast cancer cells. This specific response involving marker gene inductions by more than 1000-fold has been exploited to develop a real-time PCR-based screening method that displays a limit of detection of 5 ng g(-1) for T-2 and 10 ng g(-1) for HT-2. The practicability of this bioassay is demonstrated by its application to the detection of type A trichothecenes in different food matrices.

The effect of fungicidal treatment on selected quality parameters of barley and malt.

Protection of barley grain against contamination by fungi such as Fusarium spp., particularly by those producing mycotoxins, secondary metabolites with adverse health effects, is of principal importance. Fungicides applied immediately after full heading of spring barley is one method of direct protection. In this work, extensive two-year field experiments combined with a detailed chemical laboratory analysis (barley and malt) were performed with the aim to study the effect of previous crops, different fungicides, and other conditions on the selected barley and malt quality parameters (content of beta-glucans, pentosans, oxalic acid, deoxynivalenol, and gushing), while the main task was to follow the effect of the fungicide (used as a treatment to protect against pathogens, mostly Fusarium) on changes of the chemical composition in barley and malt, and gushing. It was found that the relationship between the studied factors and the parameters usually applied to the evaluation of barley and malt quality is quite complex and not straightforward. The responses show typical features of a multifactorial influence with both positive and negative correlations resulting in a decrease or increase in grain quality (concentrations of beta-glucans, pentosans, deoxynivalenol, and other studied parameters). The role of previous crops was also found to be important. The fungicides should be applied at the time of heading but not at the very beginning of this period.