Including non-dietary sources into an exposure assessment of the European Food Safety Authority: The challenge of multi-sector chemicals such as Bisphenol A.

In the most recent risk assessment for Bisphenol A for the first time a multi-route aggregate exposure assessment was conducted by the European Food Safety Authority. This assessment includes exposure via dietary sources, and also contributions of the most important non-dietary sources. Both average and high aggregate exposure were calculated by source-to-dose modeling (forward calculation) for different age groups and compared with estimates based on urinary biomonitoring data (backward calculation). The aggregate exposure estimates obtained by forward and backward modeling are in the same order of magnitude, with forward modeling yielding higher estimates associated with larger uncertainty. Yet, only forward modeling can indicate the relative contribution of different sources. Dietary exposure, especially via canned food, appears to be the most important exposure source and, based on the central aggregate exposure estimates, contributes around 90% to internal exposure to total (conjugated plus unconjugated) BPA. Dermal exposure via thermal paper and to a lesser extent via cosmetic products may contribute around 10% for some age groups. The uncertainty around these estimates is considerable, but since after dermal absorption a first-pass metabolism of BPA by conjugation is lacking, dermal sources may be of equal or even higher toxicological relevance than dietary sources.

Time- and temperature-dependent migration studies of Irganox 1076 from plastics into foods and food simulants.

The study provides an exhaustive set of migration data for octadecyl 3-(3,5-di-tert-butyl-4-hydroxyphenyl)propionate (Irganox 1076) from low-density polyethylene (LDPE) in several food matrices. Irganox 1076 was used as a model migrant because it represents one of the typical substances used as an antioxidant in food packaging polymers. Kinetic (time-dependent) migration studies of Irganox 1076 were performed for selected foodstuffs chosen with different physical-chemical properties and in relation to the actual European food consumption market. The effect of fat content and of the temperature of storage on the migration from plastic packaging was evaluated. The results show that migration increased with fat content and storage temperature. All data obtained from real foods were also compared with data obtained from simulants tested in the same conditions. In all studied cases, the kinetics in simulants were higher than those in foodstuffs. The work provides data valuable for the extension of the validation of migration model developed on simulants to foodstuffs themselves.

Identification and quantification of the migration of chemicals from plastic baby bottles used as substitutes for polycarbonate.

The results of a study on the analytical identification and quantification of migration of chemicals from plastics baby bottles found in the European Union market made of materials that are now present as substitutes for polycarbonate (PC) are reported. A total of 449 baby bottles with a focus on first age or sets of bottles were purchased from 26 European Union countries, Canada, Switzerland and the USA. From this collection, which contained several duplicates, a total of 277 baby bottles were analysed. The materials included different types of plastic such as PC, polyamide (PA), polyethersulphone (PES), polypropylene (PP), but also silicone, and from the United States a co-polyester marketed under the trade name Tritan™. The bottles were subjected to the conventional migration test for hot fill conditions, i.e. 2 h at 70°C. The simulant used was that specified in European Union legislation (2007/19/EC) for milk, i.e. 50% ethanol. In a first phase 1, migration was conducted since the scope of this investigation was a screening rather than a true compliance testing check. Second and third migrations were performed on selected articles when migrated substances exceeded limits specified in the legislation. In order to verify some materials, a portion of the bottle was cut to run an FT-IR fingerprint to confirm the nature of the polymer. The migration solutions in general showed a low release of substances. Results showed that bottles made of PP and silicones showed a greater number of substances in the migration solutions and in greater quantity. Chemicals from PP included alkanes, which could be found in >65% of the bottles at levels up to 3500 µg kg⁻¹; and benzene derivatives in 17% of the baby bottles and found at levels up to 113 µg kg⁻¹. Some substances were found on a regular basis such as plasticisers, esters and antioxidants (e.g. tris(2,4-di-tert-butylphenyl)phosphate, known as Irgafos 168. Some substances found were not included in the Community positive list, which means that those should not be found even in the first migration. Such substances included 2,6-di-isopropylnaphthalene (DIPN), found in 4% of the bottles at levels up to 25 µg kg⁻¹, 2,4-di-tert-butyl phenol (in 90% of the bottles at levels up 400 µg kg⁻¹). Moreover, bisphenol A (BPA) was detected and quantified in baby bottles made of PA, but limited to one brand and model specific (but labelled BPA free). Results for baby bottles made of silicone also indicated the presence of components, e.g. potentially coming from inks (benzophenone, diisopropyl naphtahalene - DIPN, which could come for example from the presence of instruction leaflets in the bottles). In the case of silicone, phthalates were also found in relevant concentrations, with levels for DiBP and DBP from the first migration test of 50-150 µg kg⁻¹ and DEHP at levels 25-50 µg kg⁻¹.

Comparison of migration from polyethersulphone and polycarbonate baby bottles.

This work presents two analytical methods developed for measuring three components of polyethersulphone (PES) and applying them to the migration testing of 30 baby bottles made of PES. The study also provides migration results under the same conditions for bisphenol A (BPA) from 40 polycarbonate baby bottles using a well-established method adapted to low concentrations. For PES bottles, migration of diphenyl sulphone (DPS), 4,4'-dichlorodiphenyl sulphone (DCPS) and 4,4'-dihydroxydiphenyl sulphone (DHPS; also known as bisphenol S) was carried out using two different analytical methods with detection limits of 0.1-0.3 µg/kg, and, therefore, much below their respective European Commission Directive 2002/72/EC legislative migration limits of 50-3000 µg/kg, respectively. In parallel, 40 bottles made of polycarbonate were analysed for the migration of BPA using a method validated at EU level and modified to give a lower detection limit of 0.1 µg/kg. Migration tests were conducted into the simulant for milk 50% EtOH (as per Commission Regulation No. 321/2011 of 1 April 2011) according to the test conditions from the guidelines on test conditions for articles in contact with foodstuffs (with a focus on kitchenware) prepared by the EU Reference Laboratory and its network of National Reference Laboratories. None of the 30 bottles made of PES released any detectable amounts of DCPS or DHPS and only two bottles released a very low amount of DPS of ~1 µg/kg in the milk food simulant compared to a regulatory limit of 3000 µg/kg. For PC bottles, 32 bottles of 40 (80%) did not release BPA above the LOD of 0.1 µg/kg (in any of the three migration tests performed on each bottle). The other 20% of bottles exhibited only very minor migration, where the highest level in the first migration test was 1.83 µg/kg and most bottles did not release detectable BPA in the second and third test. Only one bottle, with a migration level of 1.08 µg/kg, in the first test still showed a detectable level in the last migration test (i.e. 0.42 µg/kg). It is important to note that the legal limit (European Commission Directive 2002/72/EC) was still 600 µg/kg for polycarbonate bottles at the time of purchase, preceding the precautionary ban taking effect from 1 June 2011 (Commission Directive 2011/8/EU; Commission Regulation No. 321/2011). This confirms that the likelihood of migration of BPA is very low and remains at very minute amounts. The results also suggest the absence of release from PES bottles based on the set of bottles investigated.

Rapid multi-analyte quantification of benzophenone, 4-methylbenzophenone and related derivatives from paperboard food packaging.

Photo-initiators are used in printing inks hardened with UV light and one of the most commonly used photo-initiators is benzophenone (BP). Recent notifications under the Rapid Alert System for Food and Feed have shown migration of 4-methylbenzophenone (4-MBP) from packaging into cereals. A specific migration limit exists for BP of 0.6 mg kg(-1) for its use as an additive in plastics. There is no specific European legislation covering cardboard boxes and/or printing inks for food contact use. However, due to the high levels detected, the European Food Safety Authority (EFSA) published recommendations and the Standing Committee for the Food Chain and Animal Health endorsed a limit of 0.6 mg kg(-1) for the sum of BP and 4-MBP. While studies have been published on photo-initiators in the past, there is a fundamental lack of data on 4-MBP especially for its combined analysis with others. We present an HPLC method with diode array detector to simultaneously determine the levels of BP, 4-MBP as well as 7 other possible derivatives from secondary packaging for food applications. The method was tested and applied to 46 samples of paperboard for secondary packaging collected both from supermarkets and directly from a paperboard supplier. In addition, a survey was conducted on recycled paperboard (n = 19) collected from a supplier, to evaluate the background quantity of BP and other derivatives in recycled board. The most abundant photo-initiator found in the survey was BP, in 61% of samples, and 4-MBP was found in 30% of the samples. It seems that these compounds are used to replace one another. Other derivatives were found in minor quantities. Traces of BP were also found in 42% of the samples of recycled, unprinted board.

Development of a multimethod for the determination of photoinitiators in beverage packaging.

Photoinitiators are ink components used in the printing of food packaging. The aim of the present study was to develop a multimethod for the analysis of photoinitiators in paperboard packages and commercial beverage packages. The proposed method is simple and fairly rapid. Extraction is carried out with acetonitrile. A reverse-phase high performance liquid chromatography (HPLC) method with UV detection was developed to quantify the 6 selected photoionitiators. Total HPLC time analysis was 30 min and the flow-rate was 1 mL/min. Method validation parameters such as linearity and detection limit are reported. Recovery of all photoinitiators (at 1 mg/kg) from orange juice was satisfactory (mean = 88.4%; ranging from 84.6% to 93.4%). Recovery of benzophenone and Irgacure 651 was good (ranging from 85.2% to 98.1% and 86.1% to 92.2%, respectively) at the 4 concentrations studied (0.1, 0.2, 0.5, and 1 mg/kg). Commercial beverage packages were analyzed and when photoinitiators were detected, the beverages were also analyzed.

Development of an analytical method for the determination of photoinitiators used for food packaging materials with potential to migrate into milk.

Photoinitiators are used in the curing process during UV printing of food carton labels. The alarm concerning the detection of a photoinitiator, 2-isopropyl thioxanthone (ITX), in food samples packed with cartons printed with UV-cured inks has focused the attention of legislative authorities on the potential migrants from packaging inks into foods. For this reason it is very important to carry out analytical methods for the detection of those compounds in food as potential migrants from packaging. The aim of the present work was to develop a multimethod for the analysis of 6 photoinitiators in milk. The selected photoinitiators were Irgacure 184, benzophenone, Irgacure 651, Irgacure 907, Quantacure ITX, and Quantacure EHA (2-ethylhexyl-4-dimethylaminobenzoate). Milk (10 mL) extraction was carried out by using ammoniac and hexane. The supernatant was evaporated and the residue was redissolved with acetonitrile. Then, the extract was analyzed by HPLC-UV. Calibration lines were carried out over the concentration range of 0.1 to 10 mg/L. The calibration data presented high correlation coefficients (>0.9999). Mean recoveries (n = 6) of the 6 photoinitiators were 83.4% (residual standard deviation = 2.3%) at 0.5 mg/kg and 81.0% (residual standard deviation = 4.6%) at 1 mg/kg. Several milk samples and their respective packaging cartons were analyzed. Results were confirmed by HPLC-mass spectrometry.

Analytical screening of polyadipates and other plasticisers in poly(vinyl chloride) gasket seals and in fatty food by gas chromatography-mass spectrometry.

Fourteen poly(vinyl chloride) (PVC) gasket seals and 15 samples of pesto, tomatoes sauces, olive oil and olives in oil were analysed for the additives and plasticisers. The systematic screening of the representative samples revealed that epoxidised soybean oil (ESBO) was the principal plasticiser in eight gaskets and the concentrations of the substance ranged from 15% to 42%. Diisodecyl phthalate (DIDP) as main plasticiser was revealed in three samples and the amount ranged from 37 to 41%. Polyadipate was added in four samples at 16-46%. The concentration of ESBO exceeded 60 mg kg(-1) in three food samples with extreme at 281.9 mg kg(-1) and average 61.3 mg kg(-1). Di-(2-ethylhexyl) phthalate (DEHP) was detected in six food samples and it ranged from 2.5 to 8.7 mg kg(-1). The concentration of DEHP exceeded SML (3 mg kg(-1)) in five cases. The concentration of total polyadipate (PA) was 16.3 mg kg(-1) in average. All eight positive food samples for total PA showed the concentrations below SML (30 mg kg(-1)), which are specified for the fraction of polyadipates lower than 1000 Da. The migration rate of polyadipate in the food ranged from 1 to 7%. At such low transfer rate the polyadipates could be promising plasticisers for PVC gasket seals used in food industry.

Solid-phase micro-extraction (SPME-GC) and sensors as rapid methods for monitoring lipid oxidation in nuts.

Dry foods with high fat content are susceptible to lipid oxidation, which involves a quality deterioration of the product, since this process is responsible for the generation of off-flavours. Hexanal is considered to be a good shelf-life indicator of such oxidation products. In addition, due to its high volatility, hexanal can be easily determined by fast headspace analytical techniques. For this reason an electronic nose comprising ten metal oxide semiconductors (MOS) and a solid-phase microextraction (SPME) coupled with gas chromatography and flame ionization detector (GC-FID) method were compared in order to determine hexanal formed in hazelnuts during storage under different conditions (room temperature, 40 degrees C, ultraviolet light, with and without oxygen scavenger). The results obtained by the two methods showed a good correlation, confirming the possibility of using a multi-sensor system as a screening tool for the monitoring of shelf-life and oxidation state of nuts.

Survey of 2-ethylhexanoic acid in baby food.

A solid-phase microextraction method was developed and used to perform a survey of 2-ethylhexanoic acid (2-EHA) in 63 samples of baby food from 13 European Union countries, as well as from Norway and Switzerland. The levels of 2-EHA did not exceed 3.4 mg kg(-1). The concentrations of 2-EHA in 53 samples (84%) were in the range between <0.1 and 0.5 mg kg(-1). Ten samples showed the presence of the compound in the range between 0.5 and 3.4 mg kg(-1). An exposure assessment showed that the intake of 2-EHA does not exceed the tolerable daily intake (TDI) for infants of 6-12 months of age except one occurrence when taking the worst-case food consumption data available and the highest level of contamination of the survey. In the other cases, the levels of 2-EHA are at 13-fold below the TDI.

Solid-phase microextraction method for the determination of hexanal in hazelnuts as an indicator of the interaction of active packaging materials with food aroma compounds.

Fatty foods are susceptible to lipid oxidation resulting in deterioration of product quality due to the generation of off-flavours. Hexanal is a good indicator of rancidity. Therefore, a method based on solid-phase microextraction (SPME) coupled to gas chromatograph with flame ionization detection was developed to determine hexanal formation in hazelnuts during storage. Optimum conditions were as follows: carboxen-polydimethylsiloxane 75 microm fibre, extraction time 10 min, equilibrium time 10 min and equilibrium temperature 60 degrees C. The effect of oxygen scavengers on the oxidation process was also evaluated by measuring hexanal formation in hazelnuts stored with/without oxygen absorber sachets. Oxygen scavengers were shown to reduce oxidation; however, analysis of the sachet revealed that other volatile compounds from the headspace were also absorbed.

European survey of contamination of homogenized baby food by epoxidized soybean oil migration from plasticized PVC gaskets.

The results are reported of a European survey of the contamination of baby food with epoxidized soyabean oil (ESBO). Fat from the sample was extracted, transmethylated and derivatized for analysis by gas chromatography/mass spectrometry (GC/MS). Analysis was carried out of 248 samples of various types of foodstuffs from the 15 European Union Member States as well as Switzerland. ESBO was found in 95 of 248 samples analysed at levels from 1.5 to 135.2 mg kg(-1). The highest concentrations were found in main dish mixed foods such as vegetables and meat or carbohydrate- and vegetable-based mixes. Levels exceeded 30 mg kg(-1) in 15% of the samples, and 4% exceeded the overall migration limit fixed for plastics in the European Union of 60 mg kg(-1). This level of contamination is significant, confirming results from previous surveys and indicating a risk for the tolerable daily intake to be exceeded. These results suggest that an emphasis must be placed for more systematic research with a focus on the parameters for migration.

Evaluation of some screening methods for the analysis of contaminants in recycled polyethylene terephthalate flakes.

A range of different analytical techniques were used to test recycled polyethylene terephthalate (PET) flakes for potential chemical contaminants. The techniques used were headspace gas chromatography coupled to mass spectrometry (GC-MS), liquid extraction followed by GC-MS, supercritical-fluid extraction followed by GC-MS, and migration testing followed by elemental analysis using inductively coupled plasma mass spectroscopy (ICP-MS). The PET samples were 50 representative samples taken from 600 that had been collected throughout Europe and which had been screened for potential contaminants using a single technique of high-temperature GC-MS. Six of the 50 samples tested had been spiked with a selection of model contaminants, three samples were virgin PET flakes and two of the samples were from supercleaning processes. All samples were analysed 'blind' in this exercise. The qualitative results showed that most of the contaminants came from the first use, being flavour-aroma compounds from soft drinks. The quantitative analysis found concentrations under a few mg kg(-1) in the polymer, except for the spiked samples. Element migrations were low and only calcium, silicon and sodium had median migrations above 50 microg l(-1). This in-depth analysis of recycled PET flakes did not identify any significant contaminants that had not already been detected by the high-temperature static GC-MS screening method, thus demonstrating its utility.

Creation of on-line solutions in support of the dissemination of legislative and analytical information on food-contact materials.

To facilitate access to European Commission (EC) documentation and provide a public service resource of relevant information an Internet site dedicated to food-contact materials was created (http://cpf.jrc.it/webpack/) in 1998. The site contains legislative information in downloadable format, information on contacts for authorities and organizations in the field of food-contact materials, as well as information on methods for overall and specific migration. A satellite site (http://cpf.jrc.it/smt/) is dedicated to physicochemical and spectral information (FT-IR, GC-MS, NMR) as well as analytical methods on an increasing number of regulated substances. Many of these, as well as substances submitted for evaluation, are held in a centralized databank at the JRC in Ispra and aliquots can be made available to the public upon request. The site also recently expanded to provide an incentive for analytical networking between institutions by including information pages on current EC-funded and national projects and conference announcements.

Time-temperature study of the kinetics of migration of BADGE (bisphenol-A-diglycidyl-ether) into a fatty medium.

The migration kinetics of bisphenol-A-diglycidyl-ether (BADGE) from processed and non-processed model cans into vegetable oil was investigated as a function of the process treatment and the temperature of storage. Cans were either not heat-treated at all or were processed at 115 degrees C for 30 min or for 1 h after filling with oil. Each series of experiments comprised 30 samples and was further divided into three groups to be stored at different temperatures (20, 40 and 60 degrees C). Aliquots from the samples were taken at regular intervals for > 1 year. Samples were analysed for BADGE by high-performance liquid chromatography with fluorescence detection. The results showed that temperature processing had the largest effect on migration of BADGE. Storage temperature also significantly influenced migration from non-processed cans, in particular at higher storage temperatures such as 60 degrees C. Some samples were Subjected to 60 degrees C storage after an initial period at 20 degrees C and an effect on migration was also noted, although to a much lesser extent than from processing. The results of migration at higher temperatures were also correlated to the potential degradation of BADGE from oxidation products.

Occurrence of bisphenol-F-diglycidyl ether (BFDGE) in fish canned in oil.

The levels of bisphenol-F-diglycidyl ether (BFDGE) were quantified as part of a European survey on the migration of residues of epoxy resins into oil from canned fish. The contents of BFDGE in cans, lids and fish collected from all 15 Member States of the European Union and Switzerland were analysed in 382 samples. Cans and lids were separately extracted with acetonitrile. The extraction from fish was carried out with hexane followed by re-extraction with acetonitrile. The analysis was performed by reverse phase HPLC with fluorescence detection. BFDGE could be detected in 12% of the fish, 24% of the cans and 18% of the lids. Only 3% of the fish contained BFDGE in concentrations considerably above 1 mg/kg. In addition to the presented data, a comparison was made with the levels of BADGE (bisphenol-A-diglycidyl ether) analysed in the same products in the context of a previous study.

An Internet compendium of analytical methods and spectroscopic information for monomers and additives used in food packaging plastics.

An internet website (http:¿cpf.jrc.it/smt/) has been produced as a means of dissemination of methods of analysis and supporting spectroscopic information on monomers and additives used for food contact materials (principally packaging). The site which is aimed primarily at assisting food control laboratories in the European Union contains analytical information on monomers, starting substances and additives used in the manufacture of plastics materials. A searchable index is provided giving PM and CAS numbers for each of 255 substances. For each substance a data sheet gives regulatory information, chemical structures, physico-chemical information and background information on the use of the substance in particular plastics, and the food packaging applications. For monomers and starting substances (155 compounds) the infra-red and mass spectra are provided, and for additives (100 compounds); additionally proton NMR are available for about 50% of the entries. Where analytical methods have been developed for determining these substances as residual amounts in plastics or as trace amounts in food simulants these methods are also on the website. All information is provided in portable document file (PDF) format which means that high quality copies can be readily printed, using freely available Adobe Acrobat Reader software. The website will in future be maintained and up-dated by the European Commission's Joint Research Centre (JRC) as new substances are authorized for use by the European Commission (DG-ENTR formerly DGIII). Where analytical laboratories (food control or other) require reference substances these can be obtained free-of-charge from a reference collection housed at the JRC and maintained in conjunction with this website compendium.

Monitoring of bisphenol-A-diglycidyl-ether (BADGE) in canned fish in oil.

A survey at the European levels was initiated on the quantification of bisphenol-A-diglycidyl-ether (BADGE) in canned fish in oil in order to assess the exposure of BADGE. A total of 382 canned fish sample were collected from all 15 Member States and Switzerland and analysed for BADGE in fish. The fish was extracted first with hexane and reextracted with acetonitrile, followed by a membrane filtration and reverse phase HPLC analysis with fluorescence detection. The analysis of the fish showed that about 3% of the samples contained BADGE at a level above 1 mg/kg. The samples exceeding the limit by a larger margin were mostly from anchovy cans and cans manufactured in 1991-1995.

Stability testing of selected plastics additives for food contact in EU aqueous, fatty and alternative simulants.

Within the framework of the AIR3-CT94-2360 EU-project, the stability of three plastics additives in three EU aqueous and fatty food simulants and in two alternative simulants was studied under various time-temperature conditions. The additives tested were bis(2-ethylhexyl) adipate (DEHA), bis(2-ethylhexyl) phthalate (DEHP) and octadecyl 3-(3,5-di-tert-butyl-4-hydroxyphenyl) propionate (Irganox 1076). The various test conditions included exposures of 10 days at 40 degrees C, 1 h at reflux temperature for all aqueous simulants, 10 days at 40 degrees C and 1 h 175 degrees C for the olive oil and 2 days at 20 degrees C and 3 h at 60 degrees C for the isooctane simulant. Following the exposure, the additive samples were extracted from aqueous simulants with hexane. A sonication step was necessary to ensure maximum extraction of control samples. In the case of the isooctane simulant, the samples were analyzed directly from the simulant. The oil samples were extracted by acetonitrile. The extracts of samples exposed to various heat conditions as well as unexposed spiked controls and blanks were analysed by gas chromatography (GC) on a non-polar (5%--phenyl)-methylpolysiloxane capillary column with high temperature capabilities. The results showed that DEHA, DEHP and Irganox 1076 were stable at 40 degrees C and at reflux temperature in ethanolic or acidic aqueous simulants. The various additives were also stable in the organic isooctane simulant as well as in the fatty simulant olive oil. Studies on the stability of such additives used in food packaging are designed for regulatory purposes as an aid to decide whether the legislation should regulate limits for plasticizers based on a quantity in the food packaging itself or based on an ingested dose by the consumer.