The fatty acids (FA) profile was determined in n-3 enriched (Columbus™) Belgian eggs and pork in order to evaluate to what extent the n-3 fatty acids, which are very sensitive to oxidation, are resistant to storage or cooking. In standard eggs or pork, no change of the fatty acid profile was observed after storage or cooking without culinary fat, as well as in Columbus™ eggs and pork after storage. Some cooking processes (eggs in custard and meat in oven) induced a slight significant loss of n-3 fatty acids in Columbus™ eggs or pork (11.1% in fat from eggs cooked in custard vs. 15.3% in raw Columbus™ eggs and 11.0% in fat from oven cooked meat vs. 11.6% in raw Columbus™ meat). As expected, when Columbus™ pork is cooked with culinary fat, its fatty acid profile is modified according to the nature of the fat used.

Nitrofurans, banned antibiotics in the European Union (EU), have often been found in imported aquaculture products in the past and are still found nowadays according to the Rapid Alert System for Feed and Food (RASFF) of the European Commission. A quantitative method based on liquid chromatography coupled to isotopic dilution tandem mass spectrometry (LC–IDMS/MS) was developed for the determination of the residues of four nitrofuran antibiotic residues in shrimps. The experimental protocol consisted of an acid-catalysed release of protein-bound metabolites, followed by derivatisation with 2-nitrobenzaldehyde (NBA). Then, a double liquid–liquid extraction with ethyl acetate was performed before LC–IDMS/MS analysis by positive electrospray ionisation (ES+) with multiple reaction monitoring (MRM) of two transitions per compound. An “in-house” validation of the method for shrimp analysis was conducted according to the EU criteria for the analysis of veterinary drug residues in foods. The decision limits (CCalpha) were 0.08–0.36μgkg−1 and the detection capabilities (CCbeta) were 0.12–0.61μgkg−1, which are both below the minimum required performance limit (MRPL) set at 1μgkg−1 by the EU. The developed method was applied to evaluate the elimination of furazolidone residues in shrimp muscles after a contamination experiment. After 28days of decontamination, a concentration of 115μgkg−1 of furazolidone metabolite 3-amino-2-oxazolidinone (AOZ) was still measured in the shrimp muscle.

Chloramphenicol (CAP) is an antibiotic used for the treatment of bacterial infections in human and veterinary medicine. The use of CAP was prohibited in the European Union in 1994. Control laboratories are required to use suitably validated analytical methods to check sample compliance with the regulation. A quantitative method based on liquid chromatography coupled to isotopic dilution tandem mass spectrometry was developed for the determination of chloramphenicol in honey, shrimp, and poultry meat. The experimental protocol consisted of a liquid–liquid extraction with ethyl acetate. Separation and detection were realized, respectively, by a 2690 Waters HPLC (Milford, MA, USA) and a Micromass Triple Quadrupole mass spectrometer (Micromass, Manchester, UK), equipped with an electrospray source. The effects of mobile-phase additives on the response of LC/ESI/MS were examined. Two different HPLC columns were tested: the X-Terra from Waters and the Alltima HP C18 HL from Alltech (Deerfield, IL, USA). A validation of the method was conducted according to the EU criteria for the analysis of chloramphenicol in foods. The decision limits (CCα) were 0.04, 0.03, 0.07 μg kg−1, and the detection capabilities (CCβ) were 0.05, 0.04, 0.08 μg kg−1 for honey, shrimp, and poultry meat, respectively. Those values are below the minimum required performance limit set at 0.3 μg kg−1 by the EU and 0.1 μg kg−1 by Belgium. Our protocol has the advantage to propose a unique extraction method working as well for honey, shrimp, and poultry meat, contrary to similar published methods in which a different extraction method is used for each type of matrix.

Acrylamide (CH2=CHCONH2), a neurotoxic and potentially carcinogenic substance for human health, is in the glare of the spotlights for a few years. This is mostly due to the fact that acrylamide was found worldwide in various heated foodstuffs. Levels reported in the literature vary from 25 to 2000 µg/kg and potato products are considered as containing the highest level in acrylamide. A possible pathway of synthesis of acrylamide is the Maillard reaction between reducing sugars and the amino acid asparagine. The aim of this study was to develop a liquid chromatography/mass spectrometry method to analyse as quickly as possible acrylamide in a variety of Belgian food samples such as potatoes, French fries, crisp bread, coffee, corn-flakes, etc. The sample preparation consisted in a liquid/liquid extraction, a centrifugation, followed by purification with Solid Phase Extraction (SPE). The instruments used were a Waters 2690 Alliance HPLC system coupled to a Micromass Quattro Ultima Platinum triple-quadrupole mass spectrometer. The analysis was performed in MS/MS mode using isotopic dilution technique for quantification. An internal 13C3 labelled standard was added prior to extraction. Quantification in MS/MS mode was calculated by reconstructing the ion current with the most abundant daughter ions for native and 13C labelled standard (ions of m/z 55 and 58).

The objective of this study was to evaluate if the analysis results of dioxins and dioxin-like PCBs in food matrices obtained using the screening CALUX (Chemically Activated LUciferase gene eXpression) method, could be used for quantitative risk assessment.Contamination data of dioxins and dioxin-like PCBs in food matrices (dairy product, eggs, fish, animal fat and vegetable oil) obtained by the CALUX screening method and by the reference method (GC-HRMS) have been compared. Significant differences between the two methods were observed. Median concentrations of dioxins with the CALUX methods for the food matrices are about twice the median concentration of dioxins with the GC-HRMS. These discrepancies indicate that the data obtained using the CALUX method within the framework of the Belgian official control program cannot be used in risk assessment, specifically for an accurate estimation of the dietary intake of dioxins and dioxin-like PCBs. This can be explained, on one hand, by the promiscuous nature of the Aryl hydrocarbon Receptor (AhR) (whose ligand activation is responsible of the response measured in the CALUX assay) as it effectively binds to a large number of diversely shaped compounds not belonging to the 17 dioxins/furans and the 12 dioxin-like PCBs congeners. On the other hand, the assay is implemented as a screening tool for compliance monitoring where emphasis on method performance is required around the maximum levels, leading to a lack of accuracy for lower or higher concentrations.

Due to the large number of applications of bisphenol-A (BPA), the human exposure routes are multiple. We aimed to review shortly the food and non-food sources of BPA, and to evaluate their contribution to the human exposure. Food sources discussed here include epoxy resins, polycarbonate and other applications, such as paperboard and polyvinylchloride materials. Among the non-food sources, exposures through dust, thermal paper, dental materials, and medical devices were summarized. Based on the available data for these exposure sources, it was concluded that the exposure to BPA from non-food sources is generally lower than that from exposure from food by at least one order of magnitude for most studied subgroups. The use of urinary concentrations from biomonitoring studies was evaluated and the back-calculation of BPA intake seems reliable for the overall exposure assessment. In general, the total exposure to BPA is several orders of magnitude lower than the current tolerable daily intake of 50 μg/kg bw/day. Finally, the paper concludes with some critical remarks and recommendations on future human exposure studies to BPA.

The output of a pesticide surveillance program (detection frequency and number of exceeding measures) can lead to unnecessary concern among consumers since they lack information concerning the actual exposure. In this study, the exposure to pesticide residues through fruit and vegetable consumption is evaluated based on the 2008 surveillance data of the Belgian Federal Agency for the Safety of the Food Chain (FASFC). Results (deterministic and probabilistic approach) demonstrate that the chronic exposure of the adult population (>15 years) is generally under control, even at high or frequent consumption of fruit and vegetables. For most of the pesticide residues studied, the exposure is one hundred times lower than the ‘acceptable daily intake’ or ADI. With regard to children (2–5 years) who consume regularly or large amounts of fruit and vegetables, there are however, indications that for some pesticides the ADI can be exceeded. Nevertheless, due to the large uncertainty in these calculations, a more detailed study is required for this vulnerable group of consumers. In addition, it was demonstrated that washing and peeling of fruit and vegetables result in an exposure that is probably five to six times lower.

Summary A two‐plate microbiological method to screen residues of the most commonly used antibiotics in animal production, named new two‐plate test (NTPT), has been optimised and validated, according to criteria derived from the European Commission Decision 2002/657/CE. This screening method used two media at different pH seeded with a single bacteria strain ( Bacillus subtilis) . The method consists of a simple extraction, followed by the application of the extract on Petri plates. The method detected, in pork and chicken muscles, most of the antibiotics from six groups (tetracyclines, (fluoro)quinolones, penicillins, macrolides, aminoglycosides and sulfonamides) and florfenicol at concentration very close to maximum residue limits used in the EU. The screening capacity of the NTPT was compared with another screening technique, the Premi‐Test ® , by analysing spiked samples as well as real samples of meat from pork and chicken sold for local consumption, in the Red River Delta region (Vietnam). The NTPT described here appeared to detect more samples than the Premi‐Test ® , showing its interest for Vietnamese control laboratories, as a screening method to monitor antibiotic residues in chicken and pork meat, before sending the suspected samples to the confirmatory step.

Microbiological inhibition screening tests could play an important role to detect residues of antibiotics in the different animal food products, but very few are available for the aquaculture products in general, and for shrimps in particular. A two-plate microbiological method to screen shrimp for residues of the most commonly used antibiotics has been developed and validated according to criteria derived from the European Commission Decision 2002/657/CE. Bacillus subtilis was used as a sensitive strain to target antibiotics. Culture conditions on Petri plates (pH of medium) were selected to enhance the capacity of antibiotic detection. Antibiotic residues were extracted from shrimps using acetonitrile/acetone (70/30, v/v) before application on Petri plates seeded with B. subtilis. The method was validated using spiked blank tissues as well as antibiotic treated shrimps with enrofloxacin and tetracycline, two antibiotics often found to be used in shrimp production. For tetracyclines and (fluoro)quinolones, the detection capability was below the maximum residue limit (MRL), while it was around the MRL for sulfonamides. The specificity of the microbiological screening was 100% in all cases while the sensitivity and accuracy was 100% in almost all cases. The capacity of the method to detect contaminated samples was confirmed on antibiotic treated shrimps, analyzed in parallel with a confirmatory method (Liquid Chromatography coupled to mass spectrometry (LC–MS)).

Aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor mediating the adverse effects of dioxins and polycyclic aromatic hydrocarbons (PAHs). In this study, we investigated the genetic-, time-, dose-, species- and tissue-dependent AhR-mediated agonistic/antagonistic activities of three food flavonoids: quercetin, chrysin and genistein. To that end, four stably transfected cell lines were used in cell-based luciferase reporter gene assays: three lines were transformed with the ptKLuc vector harbouring four dioxin-responsive elements (DREs) upstream of the thymidine kinase promoter and the luciferase gene (HepG2-Luc, T-47D-Luc and H4IIE-ULg). The fourth is a patented cell line transformed with a different construct: H4IIE DR-CALUX®. Both H4IIE cells were compared for their genetic construction. Human hepatoma (HepG2-Luc) and human breast tumour (T-47D-Luc) cells were compared for tissue-dependent effects. Rat hepatoma (H4IIE-ULg) and human hepatoma (HepG2-Luc) cells were compared for species-dependent activities. We concluded that quercetin, chrysin and genistein act in a time-, dose-, species- and tissue-specific way. For example, genistein displayed agonistic activities when exposed to rat hepatoma cells during 6h but not after 24h. Flavonoids displayed agonistic/antagonistic activities in human breast tumour cells, depending on the exposure time, while in human hepatoma cells, only antagonistic activities of flavonoids were measured. In addition, we report, in all the cells, a synergy between an isoflavone and two food contaminants; the 2,3,7,8-tetrachlorodibenzo-p-dioxin and 3-methylcholanthrene, a PAH. In rat cells, this synergy occurred when cells were exposed to flavonoids and contaminant for 6h, while it was observed in human cells only after 24h.

The quantitative detection of allergens in the food chain is a strategic health objective as the prevalence of allergy continues to rise. Food allergenicity is caused by proteins either in their native form or in forms resulting from food processing. Progress in mass spectrometry greatly opened up the field of proteomics. These advances are now available for the detection and the quantification of traces of allergenic proteins in complex mixtures, and complete the set of biological tests used until now, such as ELISA or PCR. We review methods classified according to their ability to simultaneously quantify and identify allergenic proteins and underline major advances in the mass-spectrometric methods.