Séverine Goscinny

Sources of PCDD/Fs, non-ortho PCBs and PAHs in sediments of high and low impacted transboundary rivers (Belgium–France)

PAHs, PCDD/Fs and non-ortho PCBs have been assessed in Yser and Upper-Scheldt river sediments. Higher contamination levels were observed in the Upper-Scheldt sediments: maximum concentrations for the 16 US-EPA PAHs, PCDD/Fs and non-ortho PCBs respectively amount to 8.9 mg kg(-1), 12 ng TEQ kg(-1) and 5.1 ng TEQ kg(-1). Diagnostic PAH ratios in sediments and atmospheric samples suggest that the PAH compounds are from pyrolytic origin, more specifically combustion processes. The huge consumption of coal in cokes-ovens and smelters and its use for house-heating in Northern France, although decreasing during the last decades, are in support of that suggestion. PCDD/F fingerprints in sediments and deposition material indicate that OCDD is the dominant congener. In addition use of pentachlorophenol (PCP) in the past led to a minor contribution of PCDD/Fs in our sediment samples. Non-ortho PCBs form a substantial fraction of the total TEQ concentrations observed in the sediments. Since the 1980s and 1990s a substantial reduction of the PCDD/F sediment concentrations is observed, but this is not the case for the PAHs.

Travelling-wave ion mobility time-of-flight mass spectrometry as an alternative strategy for screening of multi-class pesticides in fruits and vegetables.

This paper reports a novel approach to screening multi-class pesticides by ion mobility time-of-flight mass spectrometry detection. Nitrogen was selected as mobility gas. After optimization of the different ion mobility parameters, determination of matrix effect on the drift times was conducted using different matrix extracts. The results showed that drift time values are not influenced by the matrix and also are independent of the concentration within the working range for 100 pesticides tested, making drift time a powerful additional identification tool. Based on statistics, 2% variation criteria provides a good fit for all the pesticides targeted, and could be considered as a maximum acceptable criteria associated with the drift time parameter for identification purpose. This 2% value is in agreement with already reported criteria, for instance, for GC or LC retention time in European documents. Finally, the well-known feature of mobility to separate complex mixtures was also tested to obtain purified extracted mass spectra of pesticides present in fruit extract.

Dietary exposure of the Belgian adult population to non-dioxin-like PCBs.

Non-dioxin-like polychlorinated biphenyls (ndl-PCBs), and some of their metabolites, might initiate neurological, neuroendocrinological, immunological and carcinogenic effects. Dietary exposure of the Belgian adult population to ndl-PCBs was investigated in this study. Foods from five food groups, collected in Belgium in 2008, were analyzed by GC-MS/MS for the six indicator PCBs (PCB 28, 52, 101, 138, 153 and 180). Results were expressed as the sum of the six congeners. A dietary exposure assessment was performed, combining ndl-PCBs levels found in food with data from the national food consumption survey of 2004. Fish and fish products were the dominating food group in terms of contamination level, with the highest levels measured in the composite sample «other fishes» (18.58 ng/g FW). The dietary exposure of the Belgian population (n=3083) to ndl-PCBs ranged from 5.33 ng/kg b.w./day on average to 16.10 ng/kg b.w./day at the 99th percentile, using the lower bound concentration. The mean dietary exposure mainly originates from Fish and fish products (54.3%), followed by dairy products (28.5%). As neither EFSA nor JECFA have set a Tolerable Daily Intake for ndl PCBs, uncertainty remains about how to interpret the exposure data in terms of public health.

Determination of caramel colorants’ by-products in liquid foods by ultra-high-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS)

2-Methylimidazole, 4-methylimidazole (2-MI and 4-MI), 2-acetyl-4-(1,2,3,4-tetrahydroxybutyl) imidazole (THI) and 5-hydroxymethylfurfural (5-HMF) are neo-formed compounds generated during the manufacture of caramel colours and are transferred to the processed food. These contaminants are known to have a toxicological profile that may pose health risks. Hence, to characterise THI, 2- and 4-MI and 5-HMF levels in liquid foods, an ultra-high-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed and sample preparation was divided into two analytical strategies depending on the concentration range expected in the type of foods targeted. For the determination of the imidazole substitutes (THI, 2- and 4-MI), a sample enrichment and clean-up step by strong cation solid-phase extraction was developed. This method is capable of quantifying over a range of 5 ng ml–1 (LOQ) to 500 ng ml–1 with recoveries of 75.4–112.4% and RSDs of 1.5–15%. For determination of 5-HMF, a standard addition method was applied covering the linear range of 0.25–30 µg ml–1 with RSDs from 2.8% (for intraday precision) to 9.2% (for intermediate precision). The validated analytical methods were applied to 28 liquid food samples purchased from local markets. THI was found only in the beer samples at levels up to 141.2 ng ml–1. For 2-MI, non-quantifiable traces were observed for all samples, while 4-MI was observed in all samples with large concentration variations (from < LOQ to 563.9 ng ml–1). 5-HMF was found at expected concentrations, except for a sherry vinegar sample (113 µg ml–1), which required a high level of dilution before following the standard addition protocol.

Low-calorie sweeteners in food and food supplements on the Italian market

This study determines the occurrence and concentration levels of artificial low-calorie sweeteners (LCSs) in food and food supplements on the Italian market. The analysed sample set (290 samples) was representative of the Italian market and comprised of beverages, jams, ketchups, confectionery, dairy products, table-top sweeteners and food supplements. All samples were analysed via UPLC-MS/MS. The method was in-house validated for the analysis of seven LCSs (aspartame, acesulfame-K, saccharin, sucralose, cyclamate, neotame and neohesperidin dihydrochalcone) in food and for five LCSs (aspartame, acesulfame-K, saccharin, cyclamate and sucralose) in food supplements. Except for cyclamate in one beverage which exceeded the maximum level (ML) with 13%, all concentrations measured in food were around or below the ML. In food supplements, 40 of the 52 samples (77%) were found to be above the ML, with exceedances of up to 200% of the ML.

Assessment of dietary intake of 10 intense sweeteners by the Italian population

The aim of the present study was to monitor the consumption of foods containing intense sweeteners present on the Italian food market and to investigate whether the Italian general population (aged >3-65+) was at risk for exceeding the Acceptable Daily Intake (ADI) of 10 intense sweeteners. A food label survey was performed in Rome (Italy), using market share data to identify the brands more representative of the market. A sample of 326 foods (table-top sweeteners included), beverages and food supplements containing intense sweeteners was collected and analyzed in order to establish the concentration levels. Intense sweeteners were only found in foods belonging to 8 sugar-free food categories out of 37 regulated. The dietary exposure was estimated using the tiered approach. Food consumption data from the last Italian national survey (INRAN-SCAI 2005-06) were combined with Maximum Levels at Tier 2, and with the actual concentration of sweeteners in the collected food products at Tier 3. The estimated exposure among consumers of sweeteners in Italy was well below the ADIs, in both tiers; non-alcoholic beverages, table-top sweeteners and food supplements were main contributors to exposure.

Occurrence of Organophosphorus Flame Retardants and Plasticizers (PFRs) in Belgian Foodstuffs and Estimation of the Dietary Exposure of the Adult Population

The occurrence of 14 organophosphorus flame retardants and plasticizers (PFRs) was investigated in 165 composite food samples purchased from the Belgian market and divided into 14 food categories, including fish, crustaceans, mussels, meat, milk, cheese, dessert, food for infants, fats and oils, grains, eggs, potatoes and derived products, other food (stocks), and vegetables. Seven PFRs [namely, tri-n-butyl phosphate (TnBP), tris(2-chloroethyl) phosphate (TCEP), tris(1-chloro-2-propyl) phosphate (TCIPP), tris(1,3-dichloro-2-propyl) phosphate (TDCIPP), triphenyl phosphate (TPHP), 2-ethylhexyldiphenyl phosphate (EHDPHP), and tris(2-ethylhexyl) phosphate (TEHP)] were detected at concentrations above quantification limits. Fats and oils were the most contaminated category, with a total PFR concentration of 84.4 ng/g of wet weight (ww), followed by grains (36.9 ng/g of ww) and cheese (20.1 ng/g of ww). Our results support the hypothesis that PFR contamination may occur during industrial processing and manipulation of food products (e.g., packaging, canning, drying, etc.). Considering the daily average intake of food for the modal adult Belgian (15-64 years of age), the dietary exposure to sum PFRs was estimated to be ≤7500 ± 1550 ng/day [103 ± 21 ng/kg of body weight (bw)/day]. For individual PFRs, TPHP contributed on average 3400 ng/day (46.6 ng/kg of bw/day), TCIPP 1350 ng/day (18.5 ng/kg of bw/day), and EHDPHP 1090 ng/day (15 ng/kg of bw/day), values that were lower than their corresponding health-based reference doses. The mean dietary exposure mainly originated from grains (39%), followed by fats and oils (21%) and dairy products (20%). No significant differences between the intakes of adult men and women were observed.

Identification of epidermal growth factor (EGF), in an unknown pharmaceutical preparation suspected to contain insulin like growth factor 1 (IGF-1).

The last 30years are marked by a tremendous evolution in the comprehension of human physiology. Although many physiological processes remain enigmatic, our increased understanding has led to a considerable development of medicines. A small, but yet a more progressively important sub fraction of this drug market consists of recombinant proteins and peptides. Although in most western countries a prescription is required for the injection of these compounds, some of them can easily be acquired from illicit internet pharmacies whether or not disguised as ‘research companies’. Some of these drugs can not only be used to cure ourselves but also to improve our sports performance, deepen our tan, and strive to the ideals imposed by society. Reports originating from controlling agencies residing in Germany, the UK, Denmark, Norway, Italy, the USA, Australia, and Belgium show that monoclonal anti‐cancer antibodies, insulins, growth hormones, human chorionic gonadotropin, Clostridium botulinum toxin type A, potential doping peptides, putative anti‐obesity drugs, skin tanning peptides, neuropeptides and a putative anti‐cancer polypeptide were identified in seized samples. The danger of these illegal polypeptides resides in the fact that these polypeptide products are not produced under controlled environment and therefore may contain the wrong ingredient, the wrong dosage, bacteria, viruses, heavy metals, etc., which could result in severe health issues. In some cases these illegal acts had a deadly outcome. Furthermore, for many of these polypeptide drugs, there is a limited knowledge of their effects and/or side‐effects (long and short term) hence clinical studies have not been terminated or even worse were given a negative advice due to overall potential adverse health effects. Global actions are on‐going to protect the sometimes unaware public from these malignant activities. In Belgium, the Federal Agency for Medicines and Health Products (FAMHP) is responsible for this task and upon their request unknown pharmaceutical preparations suspected to contain illegal drugs are regularly subject to analysis in our official medicines control laboratory (OMCL). At the end of 2014, an unknown pharmaceutical preparation, claimed to contain insulin like growth factor 1 (IGF‐1) was submitted for analysis by immunoblotting against IGF‐1 and analogues, prior to liquid chromatography‐tandem mass spectrometry (LC‐MS/MS). Based on the interpretation of the western blot and the LC‐MS data we were not able to identify this substance as being any form of insulin like growth factor (IGF), including IGF‐1, IGF‐2 or synthetic analogues potentially used as doping agent. Further analysis by LC‐MS/MS and subsequent peptide sequencing showed that the substance present in the sample corresponds to epidermal growth factor (EGF). EGF plays a major role in the regulation of cell growth, proliferation and differentiation and is therefore frequently used in cell culture. This polypeptide, consisting of 53 amino acids, with sequence NSDSECPLSHDGYCLHDGVCMYIEALDKYACNCVVGYIGERCQYRDLKWWELR and 3 intramolecular disulfide bonds has an average Molecular weight of 6215.9Da. This polypeptide is known to interact with the EGF‐receptor (EGFR) and can promote tumour cell motility and invasion. Therefore the blocking of EGF binding to the EGFR and further downstream signalling pathway is currently marked as a target for anti‐cancer therapy for certain cancers (reviewed in Yewale et al.). Indeed, a vaccine targeting EGF, named CimaVax‐EGF, results in the production of antibodies directed against EGF and this vaccine is being used as cancer therapy against non‐small‐cell lung carcinoma in Cuba. The CimaVax vaccine consists of recombinant EGF chemically conjugated to the protein P64K from Neisseria meningitides. It induces the production of antibodies directed against EGF and results in the prevention of EGF to attach to EGFR and thus inhibiting the signal that tells cancer cells to grow and divide. Nowadays, the product is undergoing further trials for possible licensing in Europe and the United States. Although no injectable legal medicine is available on the market, the product is used in the cosmetic industry. It is thought that the topical application of EGF would enhance wound healing and stimulate skin rejuvenation. Taken together, we believe that the purchaser was unaware of the presence of EGF in the sample. This case of potential dangerous wrong labelling clearly demonstrates the peril of purchasing potential doping agents via the black market.