Emerging technologies and their impact on regulatory science.

There is an evolution and increasing need for the utilization of emerging cellular, molecular and in silico technologies and novel approaches for safety assessment of food, drugs, and personal care products. Convergence of these emerging technologies is also enabling rapid advances and approaches that may impact regulatory decisions and approvals. Although the development of emerging technologies may allow rapid advances in regulatory decision making, there is concern that these new technologies have not been thoroughly evaluated to determine if they are ready for regulatory application, singularly or in combinations. The magnitude of these combined technical advances may outpace the ability to assess fit for purpose and to allow routine application of these new methods for regulatory purposes. There is a need to develop strategies to evaluate the new technologies to determine which ones are ready for regulatory use. The opportunity to apply these potentially faster, more accurate, and cost-effective approaches remains an important goal to facilitate their incorporation into regulatory use. However, without a clear strategy to evaluate emerging technologies rapidly and appropriately, the value of these efforts may go unrecognized or may take longer. It is important for the regulatory science field to keep up with the research in these technically advanced areas and to understand the science behind these new approaches. The regulatory field must understand the critical quality attributes of these novel approaches and learn from each other's experience so that workforces can be trained to prepare for emerging global regulatory challenges. Moreover, it is essential that the regulatory community must work with the technology developers to harness collective capabilities towards developing a strategy for evaluation of these new and novel assessment tools.

Regulatory landscape of nanotechnology and nanoplastics from a global perspective.

Nanotechnology and more particularly nanotechnology-based products and materials have provided a huge potential for novel solutions to many of the current challenges society is facing. However, nanotechnology is also an area of product innovation that is sometimes developing faster than regulatory frameworks. This is due to the high complexity of some nanomaterials, the lack of a globally harmonised regulatory definition and the different scopes of regulation at a global level. Research organisations and regulatory bodies have spent many efforts in the last two decades to cope with these challenges. Although there has been a significant advancement related to analytical approaches for labelling purposes as well as to the development of suitable test guidelines for nanomaterials and their safety assessment, there is a still a need for greater global collaboration and consensus in the regulatory field. Furthermore, with growing societal concerns on plastic litter and tiny debris produced by degradation of littered plastic objects, the impact of micro- and nanoplastics on humans and the environment is an emerging issue. Despite increasing research and initial regulatory discussions on micro- and nanoplastics, there are still knowledge gaps and thus an urgent need for action. As nanoplastics can be classified as a specific type of incidental nanomaterials, current and future scientific investigations should take into account the existing profound knowledge on nanotechnology/nanomaterials when discussing issues around nanoplastics. This review was conceived at the 2019 Global Summit on Regulatory Sciences that took place in Stresa, Italy, on 24-26 September 2019 (GSRS 2019) and which was co-organised by the Global Coalition for Regulatory Science Research (GCRSR) and the European Commission's (EC) Joint Research Centre (JRC). The GCRSR consists of regulatory bodies from various countries around the globe including EU bodies. The 2019 Global Summit provided an excellent platform to exchange the latest information on activities carried out by regulatory bodies with a focus on the application of nanotechnology in the agriculture/food sector, on nanoplastics and on nanomedicines, including taking stock and promoting further collaboration. Recently, the topic of micro- and nanoplastics has become a new focus of the GCRSR. Besides discussing the challenges and needs, some future directions on how new tools and methodologies can improve the regulatory science were elaborated by summarising a significant portion of discussions during the summit. It has been revealed that there are still some uncertainties and knowledge gaps with regard to physicochemical properties, environmental behaviour and toxicological effects, especially as testing described in the dossiers is often done early in the product development process, and the material in the final product may behave differently. The harmonisation of methodologies for quantification and risk assessment of nanomaterials and micro/nanoplastics, the documentation of regulatory science studies and the need for sharing databases were highlighted as important aspects to look at.

Regulatory landscape of dietary supplements and herbal medicines from a global perspective.

The number of Individuals that use dietary supplements and herbal medicine products are continuous to increase in many countries. The context of usage of a dietary supplement varies widely from country-to-country; in some countries supplement use is just limited to general health and well-being while others permit use for medicinal purposes. To date, there is little consensus from country to country on the scope, requirements, definition, or even the terminology in which dietary supplement and herbal medicines categories could be classified. Transparent science-based quality standards for the ingredients across these regulatory frameworks/definitions becomes even more important given the international supply chain. Meanwhile, there has been a rapid advancement in emerging technologies and data science applied to the field. This review was conceived at the Global Summit on Regulatory Sciences that took place in Beijing on September 2018 (GSRS2018) which is organized by Global Coalition for Regulatory Science Research (GCRSR) that consists of the global regulatory agencies from over ten countries including the European Union. This review summarizes a significant portion of discussions relating to a longitudinal comparison of the status for dietary supplements and herbal medicines among the different national jurisdictions and to the extent of how new tools and methodologies can improve the regulatory application.

Characterisation of food grade titania with respect to nanoparticle content in pristine additives and in their related food products.

Titanium dioxide is a white colourant authorised as food additive E171 in the EU and is applied in a range of food products. Currently the EU specifications for E171 do not refer to the characterisation of particle size distribution; however, this may be requested in the near future. Only a few studies have been published to date reporting data on the size distribution of food grade titanium dioxide. The aim of this study was to characterise the size distribution of titanium dioxide particles contained in eight confectionery products and the pristine titanium dioxide samples used in each of the products. This allowed the direct comparison of the particle size distribution in both the pristine and the extracted materials. By using various analytical techniques, such as transmission electron microscopy, single particle inductively coupled plasma mass spectrometry (sp-ICPMS) and centrifuge liquid sedimentation (CLS) for the characterisation and quantification of the titanium dioxide particle sizes, the impact of the instrumentation on the results was systematically studied. The volume-specific surface area (VSSA) and crystalline structure were also determined for all additives.

Managing the analytical challenges related to micro- and nanoplastics in the environment and food: filling the knowledge gaps.

This paper identifies knowledge gaps on the sustainability and impacts of plastics and presents some recommendations from an expert group that met at a special seminar organised by the European Commission at the end of 2018. The benefits of plastics in society are unquestionable, but there is an urgent need to better manage their value chain. The recently adopted European Strategy for Plastics stressed the need to tackle the challenges related to plastics with a focus on plastic litter including microplastics. Microplastics have been detected mainly in the marine environment, but also in freshwater, soil and air. Based on today's knowledge they may also be present in food products. Although nanoplastics have not yet been detected, it can be assumed that they are also present in the environment. This emerging issue presents challenges to better understand future research needs and the appropriate immediate actions to be taken to support the necessary societal and policy initiatives. It has become increasingly apparent that a broad and systematic approach is required to achieve sustainable actions and solutions along the entire supply chain. It is recognised that there is a pressing need for the monitoring of the environment and food globally. However, despite the number of research projects increasing, there is still a lack of suitable and validated analytical methods for detection and quantification of micro- and nanoplastics. There is also a lack of hazard and fate data which would allow for their risk assessment. Some priorities are identified in this paper to bridge the knowledge gaps for appropriate management of these challenges. At the same time it is acknowledged that there is a great complexity in the challenges that need to be tackled before a really comprehensive environmental assessment of plastics, covering their entire life cycle, will be possible.

Comparative, collaborative, and integrative risk governance for emerging technologies.

Various emerging technologies challenge existing governance processes to identify, assess, and manage risk. Though the existing risk-based paradigm has been essential for assessment of many chemical, biological, radiological, and nuclear technologies, a complementary approach may be warranted for the early-stage assessment and management challenges of high uncertainty technologies ranging from nanotechnology to synthetic biology to artificial intelligence, among many others. This paper argues for a risk governance approach that integrates quantitative experimental information alongside qualitative expert insight to characterize and balance the risks, benefits, costs, and societal implications of emerging technologies. Various articles in scholarly literature have highlighted differing points of how to address technological uncertainty, and this article builds upon such knowledge to explain how an emerging technology risk governance process should be driven by a multi-stakeholder effort, incorporate various disparate sources of information, review various endpoints and outcomes, and comparatively assess emerging technology performance against existing conventional products in a given application area. At least in the early stages of development when quantitative data for risk assessment remain incomplete or limited, such an approach can be valuable for policymakers and decision makers to evaluate the impact that such technologies may have upon human and environmental health.

Radical scavenging activity, anti-bacterial and mutagenic effects of cocoa bean Maillard reaction products with degree of roasting.

Raw, pre-roasted and roasted Cocoa samples were separated into four different molecular weight fractions (> 30, 30-10, 10-5 and < 5 kDa) with ultrafiltration and tested for their antibacterial, mutagenic, as well as their radical-scavenging effects. Radical-scavenging effects were tested with electro paramagnetic resonance spectroscopy, anti-mutagenicity in the Salmonella microsome assay (with and without metabolic activation), and antibacterial effects by incubating the fractions with several strains of Bifidobacteria, Enterobacter and Escherichia, and observing their growth. The radical-scavenging activity and reducing substance concentrations increased, particularly in the 5-10-kDa roasted fraction. Chromaticity testing elucidated that the 10-5-kDa fraction was one of the darkest fractions. The Salmonella microsome assay showed neither mutagenic nor anti-mutagenic effects in any of the samples at any of the different concentrations applied when using TA98, TA100 and TA102. All fractions reduced the growth of pathogenic bacteria, in particular at the highest concentration of 100 microg/mL; however, the same trends were also observed for Bifidobacteria.

Results of a European inter-laboratory comparison study on the determination of EU priority polycyclic aromatic hydrocarbons (PAHs) in edible vegetable oils.

A collaborative study on the analysis for 15 + 1 EU priority PAHs in edible oils was organised to investigate the state-of-the-art of respective analytical methods. Three spiked vegetable oils, one contaminated native sunflower oil, and one standard solution were investigated in this study. The results of 52 laboratories using either high-performance liquid chromatography with fluorescence detection or gas chromatography with mass-selective detectors were evaluated by application of robust statistics. About 95% of the laboratories were able to quantify benzo[a]pyrene together with five other PAHs included in the commonly known list of 16 US-EPA PAHs. About 80% of the participants also quantified seven additional PAHs in most samples, two of which were benzo[b]fluoranthene and benzo[k]fluoranthene, which were also known from the EPA list. Only about 50% of the participants quantified cyclopenta[cd]pyrene, benzo[j]fluoranthene, and benzo[c]fluorene. The robust relative standard deviations of the submitted results without discrimination between the methods applied ranged between 100% for 5-methylchrysene in spiked olive oil and 11% for the same analyte in spiked sunflower oil. The results clearly showed that for these analytes the methods of analysis are not yet well established in European laboratories, and more collaborative trials are needed to promote further development and to improve the performances of the respective methods.

Peanut and hazelnut traces in cookies and chocolates: relationship between analytical results and declaration of food allergens on product labels.

Accidental exposure to hazelnut or peanut constitutes a real threat to the health of allergic consumers. Correct information regarding food product ingredients is of paramount importance for the consumer, thereby reducing exposure to food allergens. In this study, 569 cookies and chocolates on the European market were purchased. All products were analysed to determine peanut and hazelnut content, allowing a comparison of the analytical results with information provided on the product label. Compared to cookies, chocolates are more likely to contain undeclared allergens, while, in both food categories, hazelnut traces were detected at higher frequencies than peanut. The presence of a precautionary label was found to be related to a higher frequency of positive test results. The majority of chocolates carrying a precautionary label tested positive for hazelnut, whereas peanut traces were not be detected in 75% of the cookies carrying a precautionary label.

Acrylamide in coffee: review of progress in analysis, formation and level reduction.

This paper summarizes the progress made in understanding the formation of acrylamide in coffee, as well as potential reduction strategies, as presented during the joint CIAA/EC workshop on acrylamide, held in Brussels in March 2006. Currently, there are no concrete measures to reduce acrylamide concentrations in roast and ground coffee without appreciably changing the organoleptic properties of the product. Certain approaches, such as steam roasting, have been tried on a laboratory scale, albeit without affording a significant reduction. More work on the mechanisms governing the "loss" of acrylamide during storage of roast and ground coffee is warranted, and studies in this direction have been initiated. Finally, risk/benefit analysis must be addressed in a complex food such as coffee, known to harbour numerous health beneficial/chemoprotective compounds with antioxidant and antimutagenic properties.

Quantification of milk fat in chocolate fats by triacylglycerol analysis using gas-liquid chromatography.

The development and in-house testing of a method for the quantification of milk fat in chocolate fats is described. A database consisting of the triacylglycerol profiles of 310 genuine milk fat samples from 21 European countries and 947 mixtures thereof with chocolate fats was created under a strict quality control scheme using 26 triacylglycerol reference standards for calibration purposes. Out of the individual triacylglycerol fractions obtained, 1-palmitoyl-2-stearoyl-3-butyroyl-glycerol (PSB) was selected as suitable marker compound for the determination of the proportion of milk fat in chocolate fats. By using PSB values from the standardized database, a calibration function using simple linear regression analysis was calculated to be used for future estimations of the milk fat content. A comparison with the widely used butyric acid method, which is currently used to determine the milk fat content in nonmilk fat mixtures, showed that both methods were equivalent in terms of accuracy. The advantage of the presented approach is that for further applications, i.e., determination of foreign fats in chocolate fats, just a single analysis is necessary, whereas for the same purpose, the C4 method requires two different analytical methods.

Determination of cocoa butter equivalents in milk chocolate by triacylglycerol profiling.

An analytical approach for the detection and quantification of cocoa butter equivalents (CBEs) in milk chocolate is presented. It is based on (i) a comprehensive standardized database covering the triacylglycerol composition of a wide range of authentic milk fat (n=310), cocoa butter (n=75), and CBE (n=74) samples and 947 gravimetrically prepared mixtures thereof, (ii) the availability of a certified cocoa butter reference material (IRMM-801) for calibration, (iii) an evaluation algorithm, which allows a reliable quantification of the milk fat content in chocolate fats using a simple linear regression model, (iv) a subsequent correction of triacylglycerols deriving from milk fat, (v) mathematical expressions to detect the presence of CBEs in milk chocolate, and (vi) a multivariate statistical formula to quantify the amount of CBEs in milk chocolate. The detection limit was 1% CBE in chocolate fat (0.3% CBE in milk chocolate, having a fat content of 30%). For quantification, the average error for prediction was 1.2% CBE in chocolate fat, corresponding to 0.4% in milk chocolate (fat content, 30%).

Analysis of phenolic acids in honeys of different floral origin by solid-phase extraction and high-performance liquid chromatography.

The determination of 18 aromatic and arylaliphatic carboxylic acids in honey from different floral origin using solid-phase extraction (SPE) and reversed-phase high performance liquid chromatography (RP-HPLC) is reported. The behaviour of the solutes on SPE cartridges was predicted from preliminary calculations involving the pK(a) constants of the carboxylic groups, the n-octanol:water partition coefficients and the distribution coefficients at different pH values of the conditioning and washing solvents. The proposed SPE isolation and pre-concentration of the acids was achieved on reversed-phase Bond Elut C18 cartridges using an acetonitrile:tetrahydrofuran (1:1, v/v) elution system. RP-HPLC separations were performed on a Spherisorb ODS-2 column using linear gradient elution with a mobile phase composed of 20 mm phosphate buffer (pH 2.92) and methanol, and with UV detection. The reported SPE and RP-HPLC methods were applied to the analysis of 49 authentic honey samples from various floral sources and the results indicate that they may serve with respect to the quantitative control of a number of phenolic acids in plant-derived foods and medicinal plants.

Collaborative trial validation study of two methods, one based on high performance liquid chromatography-tandem mass spectrometry and on gas chromatography-mass spectrometry for the determination of acrylamide in bakery and potato products.

A European inter-laboratory study was conducted to validate two analytical procedures for the determination of acrylamide in bakery ware (crispbreads, biscuits) and potato products (chips), within a concentration range from about 20 microg/kg to about 9000 microgg/kg. The methods are based on gas chromatography-mass spectrometry (GC-MS) of the derivatised analyte and on high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) of native acrylamide. Isotope dilution with isotopically labelled acrylamide was an integral part of both methods. The study was evaluated according to internationally accepted guidelines. The performance of the HPLC-MS/MS method was found to be superior to that of the GC-MS method and to be fit-for-the-purpose.

Method validation for determination of the 15 European-priority polycyclic aromatic hydrocarbons in primary smoke condensates by gas chromatography/mass spectrometry: interlaboratory study.

A collaborative study was conducted to validate an analytical method for quantification of the 15 polycyclic aromatic hydrocarbons (PAHs) regarded in 2002 as a health concern by the former Scientific Committee on Food of the European Commission (SCF) in primary smoke condensates. The method is based on gas chromatography/mass spectrometry of a cyclohexane extract with solid-phase cleanup through silica gel. The analytes were detected in the selected-ion monitoring mode and quantified by using 3 isotopically labeled internal standard compounds. Seventeen laboratories participated in the collaborative validation study, of which 12 reported valid results. The data were subjected to Cochran, single Grubbs, and double Grubbs tests for statistical outliers. A maximum of 2 outliers was eliminated before further statistical evaluation of the method performance characteristics. Depending on the analyte, the results showed relative standard deviations for repeatability between 4.2 and 30% and for reproducibility from 9.9 to 60%. The recoveries varied between about 50 and 85%, except those for cyclopenta[cd]pyrene, dibenzo[a,l]pyrene, and dibenzo[a,h]pyrene. Nevertheless, because Commission Directive 2005/10/EC allows for a recovery range of 50-120% for (BaP) benzo[a]pyrene in various foods, it can be concluded that the method performs appropriately within the analytical range between 5 and 25 microg/kg of primary smoke condensate. For BaP the validated analytical range covered 5-20 microg/kg, and for benzo[a]anthracene (BaA) 10-25 microg/kg. The method is suitable for monitoring BaP and BaA at their respective maximum permitted levels of 10 and 20 microg/kg. Three analytes, benzo[b]-, benzo[j]-, and benzo[k]-fluoranthene could not be separated by all of the participants and were therefore treated as the sum. Nevertheless, with this method the pattern of the respective concentrations of these 15 PAHs can be monitored in primary smoke condensate as suggested by the SCF.

Validation of two commercial lateral flow devices for the detection of peanut proteins in cookies: interlaboratory study.

Results are reported for an interlaboratory validation study of 2 commercially available Iateral flow devices (dipstick tests) designed to detect peanut residues in food matrixes. The test samples used in this study were cookies containing peanuts at 7 different concentrations in the range of 0-30 mg peanuts/kg food matrix. The test samples with sufficient and proven homogeneity were prepared in our laboratory. The analyses of the samples (5 times per level by each laboratory) were performed by 18 laboratories worldwide which submitted a total of 1260 analytical results. One laboratory was found to be an outlier for one of the test kits. In general, both test kits performed well. However, some false-negative results were reported for all matrixes containing < 21 mg peanuts/kg cookie. It must be stressed that the test kits were challenged beyond their cut-off limits (> or = 5 mg/kg, depending on the food matrix). One test kit showed fewer false-negative results, but it led to some false-positive results for the blank materials. The sensitivity of the dipstick tests approaches that achieved with enzyme-linked immunosorbent assays.