UV-filters and musk fragrances in seafood commercialized in Europe Union: Occurrence, risk and exposure assessment.

In the framework of the FP7 ECsafeSeafood project, 62 seafood samples commercialized in Europe Union from several representative species - mackerel, tuna, salmon, seabream, cod, monkfish, crab, shrimp, octopus, perch and plaice - were analysed for residues of 21 personal care products (PCPs), including 11 UV-filters (UV-Fs) and 10 musk fragrances (musks). PCPs analysis were performed by Quick, Easy, Cheap, Effective Rugged, Safe (QuEChERS), combined with liquid-liquid extraction (LLE) or dispersive solid-phase extraction (dSPE), followed by gas chromatography-tandem mass spectrometry (GC-MS/MS). The results showed the presence in a wide range of samples of nine out of eleven UV-Fs compounds analysed, namely 2-ethylhexyl salicylate (EHS), 2-ethylhexyl,4-methoxycinnamate (EHMC), 4-methylbenzylidenecamphor (4-MBC), benzophenone-1 (BP1), benzophenone-3 (BP3), isoamyl-4-methoxycinnamate (IMC), 2,2'-dihydroxy-4,4'-dimethoxybenzophenone (DHMB), homosalate (HS), and octocrylene (OC), whereas galaxolide (HHCB), galaxolide lactone (HHCB-lactone), and tonalide (AHTN) were the most found musks. The potential risks to human health associated with the exposure to eight of the more prevalent PCPs - EHS, EHMC, 4-MBC, BP1, BP3, IMC, HHCB, and AHTN - through seafood consumption were assessed for consumers from five European countries (Belgium, Ireland, Italy, Portugal and Spain). Results showed that the human exposure to UV-Fs and musks estimated from the concentration values found in seafood and the daily consumption of concerned seafood species, were far below toxicological reference values.

Recycling of plastic waste: Screening for brominated flame retardants (BFRs).

Flame retardants are chemicals vital for reducing risks of fire and preventing human casualties and property losses. Due to the abundance, low cost and high performance of bromine, brominated flame retardants (BFRs) have had a significant share of the market for years. Physical stability on the other hand, has resulted in dispersion and accumulation of selected BFRs in the environment and receiving biota. A wide range of plastic products may contain BFRs. This affects the quality of waste plastics as secondary resource: material recycling may potentially reintroduce the BFRs into new plastic product cycles and lead to increased exposure levels, e.g. through use of plastic packaging materials. To provide quantitative and qualitative data on presence of BFRs in plastics, we analysed bromophenols (tetrabromobisphenol A (TBBPA), dibromophenols (2,4- and 2,6-DBP) and 2,4,6-tribromophenol (2,4,6-TBP)), hexabromocyclododecane stereoisomers (α-, ß-, and γ-HBCD), as well as selected polybrominated diphenyl ethers (PBDEs) in samples of household waste plastics, virgin and recycled plastics. A considerable number of samples contained BFRs, with highest concentrations associated with acrylonitrile butadiene styrene (ABS, up to 26,000,000ngTBBPA/g) and polystyrene (PS, up to 330,000ng∑HBCD/g). Abundancy in low concentrations of some BFRs in plastic samples suggested either unintended addition in plastic products or degradation of higher molecular weight BFRs. The presence of currently restricted flame retardants (PBDEs and HBCD) identified in the plastic samples illustrates that circular material flows may be contaminated for extended periods. The screening clearly showed a need for improved documentation and monitoring of the presence of BFRs in plastic waste routed to recycling.

Dietary exposure to volatile and non-volatile N-nitrosamines from processed meat products in Denmark.

Recent epidemiological studies show a positive association between cancer incidence and high intake of processed meat. N-nitrosamines (NAs) in these products have been suggested as one potential causative factor. Most volatile NAs (VNAs) are classified as probable human carcinogens, whereas the carcinogenicity for the majority of the non-volatile NA (NVNA) remains to be elucidated. Danish adults (15-75 years) and children (4-6 years) consume 20 g and 16 g of processed meat per day (95th percentile), respectively. The consumption is primarily accounted for by sausages, salami, pork flank (spiced and boiled) and ham. This consumption results in an exposure to NVNA of 33 and 90 ng kg bw(-1) day(-1) for adults and children, respectively. The exposure to VNA is significantly lower amounting to 0.34 and 1.1 ng kg bw(-1) day(-1) for adults and children, respectively. Based on a BMDL10 of 29 µg kg bw(-1) day(-1) a MOE value ≥17,000 was derived for the exposure to NA known to be carcinogenic (VNA including NSAR), indicating an exposure of low concern. The exposure to the NVNA is substantially higher and if found to be of toxicological significance the exposure may be of concern.

Formation and mitigation of N-nitrosamines in nitrite preserved cooked sausages.

Literature on formation and mitigation of N-nitrosamine (NA) and especially non-volatile NA (NVNA) in meat products is scarce and the present study is therefore a relevant contribution to the field. We found positive correlation between the levels of N-nitrosopiperidine (NPIP), N-nitrosohydroxyproline (NHPRO), N-nitrosoproline (NPRO), N-nitrosothiazolidine-4-carboxylic acid (NTCA) and N-nitroso-2-methyl-thiazolidine-4-carboxylic acid (NMTCA) and the amount of nitrite added to cooked pork sausages. The levels studied were 0, 60, 100, 150, 250 and 350 mg kg(-)(1). The levels of N-nitrosodimethylamine (NDMA) and N-nitrosopyrrolidine (NPYR) remained at or below limit of quantification. Erythorbic acid inhibited the formation of NHPRO, NPRO, NPIP and NTCA. This inhibition was for NTCA and NMTCA counteracted by addition of free iron. Ascorbyl palmitate had less inhibitory effect than erythorbic acid and a combination of the two provided no further protection. Increasing the black pepper content increased the levels of NPIP and NMTCA. Only slight effects of increased fat content and addition of tripolyphosphate were observed.

Simultaneous determination of volatile and non-volatile nitrosamines in processed meat products by liquid chromatography tandem mass spectrometry using atmospheric pressure chemical ionisation and electrospray ionisation.

A sensitive, selective and generic method has been developed for the simultaneous determination of the contents (µgkg(-1) range) of both volatile nitrosamines (VNA) and non-volatile nitrosamines (NVNA) in processed meat products. The extraction procedure only requires basic laboratory equipment and a small volume of organic solvent. Separation and quantification were performed by the developed LC-(APCI/ESI)MS/MS method. The method was validated using spiked samples of three different processed meat products. Satisfactory recoveries (50-130%) and precisions (2-23%) were obtained for eight VNA and six NVNAs with LODs generally between 0.2 and 1µgkg(-1), though for a few analyte/matrix combinations higher LODs were obtained (3 to 18µgkg(-1)). The validation results show that results obtained for one meat product is not always valid for other meat products. We were not able to obtain satisfactory results for N-nitrosohydroxyproline (NHPRO), N-nitrosodibenzylamine (NDBzA) and N-nitrosodiphenylamine (NDPhA). Application of the APCI interface improved the sensitivity of the method, because of less matrix interference, and gave the method a wider scope, as some NAs were ionisable only by APCI. However, it was only possible to ionize N-nitroso-thiazolidine-4-carboxylic acid (NTCA) and N-nitroso-2-methyl-thiazolidine-4-carboxylic acid (NMTCA) by ESI. The validated method was applied for the analysis of processed meat products and contents of N-nitrosodimethylamine (NDMA), N-nitrosopyrrolidine (NPYR), N-nitrosomethylaniline (NMA), N-nitrosoproline (NPRO), NTCA, and NMTCA were found in one or several nitrite cured meat products, whereas none were detected in non-nitrite cured bacon.

Influence of smoking parameters on the concentration of polycyclic aromatic hydrocarbons (PAHs) in Danish smoked fish.

A new method for the analysis of 25 polycyclic aromatic hydrocarbon (PAH) compounds in fish was developed, validated, and used for the quantification of PAHs in 180 industrially smoked fish products. The method included pressurized liquid extraction, gel-permeation chromatography (Bio-beads S-X3), solid-phase extraction (silica gel), and gas chromatography-mass spectrometry analysis. The sum concentration of 25 PAHs ([summation operator]PAH(25)) was highest in smoked herring (n = 3) and mackerel fillets (n = 13), with an average concentration of 320 and 235 microg kg(-1), respectively. Lowest average [summation operator]PAH(25) concentrations were obtained for indirectly smoked trout (26 microg kg(-1)). Principal component analysis was used to correlate processing parameters to PAH concentrations and to identify the effects of these parameters. The analysis showed that for salmon hot-smoking conditions lead to higher SigmaPAH(25) than cold smoking, and for other fish species direct smoking leads to higher SigmaPAH(25) than indirect smoking. Also, the usage of common alder increases the PAH contamination compared with beech. The effects of smoking time, combustion temperatures, and two types of smoke-generating material on the [summation operator]PAH(25) were also tested in a pilot plant study with smoked trout as a model fish. In addition to confirming that increased combustion temperatures and usage of common alder in comparison with beech increased [summation operator]PAH(25), it was also revealed that the PAH concentration decreased in the order fish skin >> outer layer of the fish muscle > inner part of the fish muscle.

Analysis of glyphosate residues in cereals using liquid chromatography-mass spectrometry (LC-MS/MS).

A fast and specific method for the determination of glyphosate in cereals is described. The method is based on extraction with water by ultrasonication. The samples are cleaned up and separated by high-performance liquid chromatography on a polystyrene-based reverse-phase column (clean-up) in series with an ion chromatography column (separation) using NaHCO(3) as eluent. A micro-membrane suppressor was inserted after the separator column to remove the Na(+) ions before detection by electrospray ionization mass spectrometry in the negative-ion mode. In MS/MS, mode the following transitions were monitored m/z 168--> 150 (glyphosate) and 170-->152 (internal standard 2-(13)C(15)N-glyphosate) for quantification. The mean recovery was 85% (n=32) at spiking levels from 0.03 to 0.33 mg kg(-1). From 1998 to 2001, from the analysis of about 50 samples per annum, a reduction in the glyphosate residues was observed owing to a Danish trade decision not to use grain with glyphosate residues for milling or bread production.

Processing factors and variability of pyrimethanil, fenhexamid and tolylfluanid in strawberries.

An HPLC-MS/MS method for the analysis of three pesticides in strawberries was developed and validated. Recoveries were measured at three spiking levels and ranged from 85 to 99% (mean recoveries). The effects of processing of strawberries ranging from rinsing to jam production were investigated for the three fungicides tolylfluanid, fenhexamid and pyrimethanil, which were applied under field conditions. Kresoxim-methyl was also applied in the field, but was not found in any of the samples investigated. The effect of parameters such as preharvest interval, dose, harvest time and observed pesticide concentration after harvest (initial concentration, mg kg(-1)), were examined with respect to possible reduction of the pesticides. The results from rinsing showed that all three pesticides were reduced on average by 37% for tolylfluanid, by 34% for fenhexamid and by 19% for pyrimethanil. For tolylfluanid and fenhexamid, the initial concentration significantly affected the reduction. For fenhexamid, dose could also have a minor influence on reduction. For pyrimethanil, none of the parameters significantly influenced the reduction. For jam production, cooking significantly reduced tolylfluanid by an average of 91%. For fenhexamid and pyrimethanil, a smaller reduction was seen, 25% and 33%, respectively. The reduction of tolylfluanid and pyrimethanil was affected by the preharvest interval, while fenhexamid was affected by the initial concentration. The unit-to-unit variability of fungicide contents was also investigated and the variability factors for the three fungicides were from 1.9 to 2.8.

Cumulative risk assessment of the intake of organophosphorus and carbamate pesticides in the Danish diet.

The aim of the study is to evaluate the potential cumulative effects of organophosphorus and carbamate pesticides that act through a common mechanism of toxicity, and to assess the long- and short-term risks for the Danish population. The intake estimates are based on dietary intake data collected in the Danish nation-wide food consumption survey in 1995. The pesticide data are based on the Danish pesticide residue-monitoring programme from 1996-2001. The amount of 35 organophosphorus pesticides and carbamates were included in the cumulative risk assessment. Processing factors, such as reduction of pesticide levels by rinsing and peeling, were applied in the exposure assessment. The "Toxicity Equivalence Factor" (TEF) approach was used to normalise the toxicity of the different organophosphorus and carbamate pesticides. Cumulative chronic exposure of organophosphorus and carbamates pesticides via fruit, vegetables and cereals is for adults 0.8-2% of the Acceptable Daily Intake (ADI) in chlorpyrifos equivalents, and 0.03-11% of the ADI in methamidophos equivalents; and for children 2-5% of the ADI in the chlorpyrifos equivalents, and 0.07-27% of the ADI in methamidophos equivalents. Neither Acute Reference Dose (ARfD) nor ADI was exceeded for any of the compounds studied. The results indicate that the Danish population is neither exposed to any cumulative chronic risk, nor at risk of acute exposure, from consumption of organophosphorus and carbamate pesticides from fruit, vegetables and cereals.

Method validation for strobilurin fungicides in cereals and fruit.

Strobilurins are a new class of fungicides that are active against a broad spectrum of fungi. In the present work a GC method for analysis of strobilurin fungicides was validated. The method was based on, extraction with ethyl acetate/cyclohexane, clean-up by gel permeation chromatography, (GPC) and determination of the content by gas chromatography (GC) with electron capture (EC-), nitrogen/phosphorous (NP-), and mass spectrometric (MS-) detection. Three strobilurins, azoxystrobin, kresoxim-methyl and trifloxystrobin were validated on three matrices, wheat, apple and grapes. The validation was based on recovery tests at three or four spiking levels, determined as double determinations and repeated three times (n = 6). The mean recoveries for the three detectors were in the range of 70-114%, and the LODs were between 0.004 mg/kg and 0.014 mg/kg, for all three strobilurins. The values for repeatability and reproducibility were within the limits for repeatability and reproducibility given by the Horwitz equation. Validation was not accepted for azoxystrobin in grapes on all three detectors and for azoxystrobin in apple for the MS-detector. A comparison of matrix-matched standards versus standards in solvent showed varying differences between the two calibration curves.

Investigation of the herbicide glyphosate and the plant growth regulators chlormequat and mepiquat in cereals produced in Denmark.

An LC-MS/MS method for analysing glyphosate and aminomethylphosphonic acid (AMPA) in cereals was developed. The method is based on extraction with water and detection of the ions from the fragmentation m/z 170-->88 (glyphosate) and m/z 112-->30 (AMPA), using electrospray interface in the positive mode. Investigation from the harvests of 1998 and 1999 showed residues of glyphosate and/or its degradation product AMPA in more than half of the cereal samples produced in Denmark. The average concentration of glyphosate in 46 samples from the 1999 harvest was 0.11 mg/kg compared with 0.08 mg/kg for the 1998 harvest (n = 49). Thus, the figures were well below the maximum residue limit (MRL) and no violations were observed. The plant growth regulators chlormequat and/or mepiquat were investigated in cereals from the Danish harvest of 1999 where 83% of the samples contained chlormequat (n = 46) compared with 87% of the samples from the 1997 harvest (n = 52). The average concentration of chlormequat in 1999 was 0.32 mg/kg compared with 0.23 mg/kg in 1997. At 2.9 mg/kg, one sample of wheat bran was exceeding the MRL of 2 mg/kg for wheat. The intakes of the pesticides through the diet of cereals were estimated to comprise 0.04% of the acceptable daily intake (ADI) for glyphosate and 1% of the ADI for chlormequat for an adult Dane.