MICROPLASTICS: Focus on Food and Health

Every year worldwide, more than 300 million tons of plastics are produced, half of which is designed for single use, and each year, at least 8 million tons end up in our oceans. These plastics break down with the intervention of microbes into carbon dioxide, methane and water, but this process is temperature dependent and readily occurs in the marine environment. Furthermore, the polymers most commonly used (e.g. PE, PP, PVC) are not readily biodegradable; they are subjected to weathering and fragmenting into micro- and nano-plastics and remain in the environment for hundreds of years. These small particles of plastic can be ingested by zooplankton, invertebrates and small fish, entering this way in the food-chain.JRC.F.7-Knowledge for Health and Consumer Safet

The certification of mass concentration of Beta-2-microglobulin in human serum: ERM-DA470k/IFCC

This report describes the additional certification of the mass concentration of Beta-2-microglobulin (B2M) in ERM-DA470k/IFCC, a human serum material. The material was certified following ISO Guide 34:2009. The material was released in 2008 and was certified for the mass concentration of 12 proteins in human serum. A full description of the processing steps can be found in the original report. Between unit-homogeneity was quantified and stability during dispatch and storage were assessed in accordance with ISO Guide 35:2006. Within-unit homogeneity was estimated to determine the minimum sample intake. The material was characterised by an intercomparison among laboratories of demonstrated competence and adhering to ISO/IEC 17025. Uncertainties of the certified values were calculated in compliance with the Guide to the Expression of Uncertainty in Measurement (GUM) and include uncertainties related to possible inhomogeneity, to instability and to characterisation. The material is intended for the calibration of immunoassay-based in-vitro diagnostic devices or control products for the proteins certified. As for any calibrator it should be verified that it is commutable. The material is produced in a similar manner as ERM-DA470, the use of which has led to a significant reduction in the between-method and between-laboratory variation for the proteins certified (B2M was not certified in this material) [ , ]. It was verified during the value assignment procedure that there were no significant matrix effects, and that different methods produced consistent results. However, when the material is used as a calibrator, the commutability should be verified for the particular assay concerned. The Certified Reference Material (CRM) is available in the lyophilised form of a 1.0 mL portion of serum with additives (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), sodium azide, benzamidine hydrochloride, sodium chloride and aprotinin). The material is kept under nitrogen gas in threaded glass bottles with rubber stoppers and polypropylene screw caps. The water mass fraction of the sample is (4.3 ± 0.6) mg/g. The lyophilised powder has to be reconstituted with (1.00 ± 0.01) g of distilled water. The minimum amount of reconstituted sample to be used is 2 µL. The CRM was accepted as European Reference Material (ERM®) after peer evaluation by the partners of the European Reference Materials consortium.JRC.D.2-Standards for Innovation and sustainable Developmen

China: Challenges and Prospects from an Industrial and Innovation Powerhouse

China is rapidly becoming a major industrial competitor in high tech and growth sectors. Its economic success and related industrial policies have received a high degree of attention, especially in light of its capacity to challenge the leading position of advanced economies in several fields. China aims, through the 'Made in China 2025' strategy, to become a world leader in key industrial sectors. In these sectors, it strives to strengthen its domestic innovation capacity, to reduce its reliance on foreign technologies while moving up in global value chains. This report analyses China's approach to attain a dominant position in international markets through a combination of industrial, R&I, trade and foreign direct investment policies. It offers an assessment of China's current position compared to the EU and US innovation systems across a range of dimensions. It concludes that China has become a major industrial competitor in several rapidly expanding high tech sectors, which may well result in attaining China's goal of becoming an innovation leader in specific areas. As a response, the EU will need to boost its industrial and R&I performance and develop a trade policy that can ensure a level playing field for EU companies in China and for Chinese companies in the EU.JRC.B.7-Knowledge for Finance, Innovation and Growt

Fipronil in eggs: Factsheet – December 2017

News about the so-called 'FIPRONIL case' has received widespread media coverage, causing one of the most significant food scares in Europe since the 2013 'horse meat scandal' and leading to a judiciary enquiry and withdrawal of millions of eggs from supermarket shelves. The 'FIPRONIL case' is linked to the discovery of the insecticide in contaminated eggs and egg products in several EU Member States and outside Europe. FIPRONIL is authorised to be used as veterinary medicine to combat fleas, mites and ticks in dogs and cats but forbidden for use in animals that are intended for the food chain, such as chickens. The European Commissioner for Health and Food Safety, Vytenis Andriukaitis, communicated that 26 of the 28 EU Member States reported the presence of FIPRONIL in eggs and egg products, with more than 45 countries affected worldwide, including the United States, Russia, Israel and Canada. To avoid contaminated food products entering into the food chain, food safety authorities of EU Member States ensure that eggs, egg products or chicken meat from affected farms are neither placed on the EU market nor exported to non EU-countries. In order to enssure adequate control, the Joint Research Centre (JRC) was requested to offer support in assessing the competence of the official laboratories in quantifying FIPRONIL as specified in the EU legislation.JRC.F.7-Knowledge for Health and Consumer Safet

'FAKE RICE' on African and Asian Markets. Rumour of Evidence?: Factsheet – December 2017

In June 2017, local journals and social media warned about the presence of plastic rice on the markets of Ghana, Nigeria and India. Similar rumours have been circulating in the press in Asia since 2016. This rice is made by mixing sweet potatoes and a polymer. But other versions also mention plastic pellets being mixed with normal rice. This type of falsifications may cause gastritis and other stomach related diseases. However, official controls could not confirm the presence of plastic rice.JRC.F.7-Knowledge for Health and Consumer Safet

Quantification of Protein Calibrants by Amino Acid Analysis Using Isotope Dilution Mass Spectrometry

This work demonstrates that amino acid analysis based on isotope dilution mass spectrometry (IDMS) can be applied to quantify proteins having different complexities and natures. Five proteins and one decapeptide were selected for the study: C-reactive protein (CRP), beta-2-microglobulin (B2 M), cystatine C (CysC), human serum albumin (HSA), Ara h1, and angiotensin I. The quantification was based on the determination of four amino acids, proline (Pro), isoleucine (Ile), valine (Val), and phenylalanine (Phe) within a working range between 5 and 100 pmol/injection of each amino acid, after 60 min digestion with HCl at 150 degrees C. The amino acids were selected taking into account their abundance in the protein sequence and to include the more difficult to break peptide bonds. Quantification of the protein amounts calculated from each amino acid is consistent, indicating that the method is working reliably. This consistency points to a complete hydrolysis of the proteins. The trueness of the method was proven when dry mass determination after dialysis was applied to HSA and CRP and the results were compared to those from amino acid analysis. Traceability to SI was assured by extensive characterisation of the amino acid calibrants by nuclear magnetic resonance, neutron activation analysis, and Karl Fischer titration.JRC.D.2-Reference material

Extreme DNA Bending: Molecular Basis of the Regulatory Breadth of IHF (Chapter 16)

The Integration host factor (IHF) is a heterodimeric, sequence-specific DNA-binding and DNA-bending protein found in many types of eubacteria. The sole function of IHF is to bring about a sharp curvature in the target DNA (up to = 160°). Such a drastic change in DNA shape has been evolutionarily recruited for controlling a large number of functions that depend on the architecture of given genomic sites. These include the organization of the bacterial nucleoid and the transcriptional control of distinct promoters. The growing availability of bacterial genomes allows a comparative approach to survey the regulatory breadth of IHF in a wider context. In this Chapter, we use the sequence of the IHF protein of the soil bacterium Pseudomonas putida as a starting point to examine in detail the basis of the recognition of DNA sequences by this nucleoid-associated protein, in particular the correlation between sequence conservation and DNA interaction for each of the IHF chains. This is greatly facilitated by comparing the protein sequence and the DNA binding specificity of IHF with those of similar proteins HU and the transcription factor 1 (TF1) from bacteriophage SPO1 of Bacillus subtilis. Mapping of the fully conserved amino acids and the protein-specific sites for each chain of the corresponding tridimensional structures finely correlates with those sites involved in DNA interactions and maintaining the protein dimer structure. The sequence conservation profile of the DNA-binding regions of these proteins shows that chain B of IHF is more closely related to HU/TF1 than to chain A of IHF, suggesting a separate evolutionary origin. Furthermore, some features of the DNA recognition mechanism seem to be exclusive to IHF and cannot be fulfilled by HU or TF1 proteins.HU and TF1 can be embraced by DNA as IHF can by the action of residues conserved in the three proteins (thereby explaining why HU/TF1 and IHF can be partially replaced by each other). In contrast, only the interactions mediated by tree-determinants (i.e. those residues that are specific for each chain of IHF) can afford a high DNA recognition specificity. These analyses highlight the importance of DNA binding versus DNA bending specificities for expansion of the regulatory space of such nucleoid-associated proteins.JRC.DG.D.2-Reference material

Development and optimisation of a generic micro LC-ESI-MS method for the qualitative and quantitative determination of 30-mer toxic gliadin peptides in wheat flour for food analysis.

We sometimes see manufactured bakery products on the market which are labelled as being gluten free. Why is the content of such gluten proteins of importance for the fabrication of bakery industry and for the products? The gluten proteins represent up to 80 % of wheat proteins, and they are conventionally subdivided into gliadins and glutenins. Gliadins belong to the proline and glutamine-rich prolamin family. Its role in human gluten intolerance, as a consequence of its harmful effects, is well documented in the scientific literature. The only known therapy so far is a gluten-free diet, and hence, it is important to develop robust and reliable analytical methods to quantitatively assess the presence of the identified peptides causing the so-called coeliac disease. This work describes the development of a new, fast and robust micro ion pair-LC-MS analytical method for the qualitative and quantitative determination of 30-mer toxic gliadin peptides in wheat flour. The use of RapiGest™ SF as a denaturation reagent prior to the enzymatic digestion showed to shorten the measuring time. During the optimisation of the enzymatic digestion step, the best 30-mer toxic peptide was identified from the maximum recovery after 3 h of digestion time. The lower limit of quantification was determined to be 0.25 ng/μL. The method has shown to be linear for the selected concentration range of 0.25–3.0 ng/μL. The uncertainty related to reproducibility of measurement procedure, excluding the extraction step, has shown to be 5.0 % (N = 12). Finally, this method was successfully applied to the quantification of 30-mer toxic peptides from commercial wheat flour with an overall uncertainty under reproducibility conditions of 6.4 % including the extraction of the gliadin fraction. The results were always expressed as the average of the values from all standard concentrations. Subsequently, the final concentration of the 30-mer toxic peptide in the flour was calculated and expressed in milligrams per gram unit. The determined, calculated concentration of the 30-mer toxic peptide in the flour was found to be 1.29 ± 0.37 μg/g in flour (N = 25, s y  = 545,075, f = 25 − 2 (t = 2.069), P = 95 %, two-sided).JRC.F.7-Knowledge for Health and Consumer Safet

CERTIFICATION REPORT Certification of the amount-of-substance fraction of HbA1c versus the sum of HbA0 and HbA1c in haemoglobin isolated from whole blood: ERM®-AD500/IFCC

This report describes the production of ERM-DA500/IFCC, which is a set of 6 ampoules containing haemoglobin with different levels of HbA1c, certified for the amount-of-substance fraction of HbA1c versus the sum of HbA0 and HbA1c. HbA1c is defined as the stable adduct from glucose and the N-terminal amino group of the β-chain of haemoglobin A0 that is beta-N-(1-deoxyfructos-1-yl) haemoglobin. This material was produced following ISO Guide 34:2009 [ ] and is certified in accordance with ISO Guide 35:2006 The ampoules contain buffered solutions of haemoglobin with different amount of substance concentrations of HbA1c and HbA0. The base materials (solutions of purified HbA1c and HbA0) were prepared from whole blood obtained from diabetic volunteers and provided by Roche Diagnostic GmbH, Department for Biochemical Materials, Penzberg, Germany. Mixtures of these base materials were filled into the ampoules, with each ampoule containing approximately 1 mg haemoglobin in 30 microliter solution, sealed under an atmosphere of argon. Between-unit homogeneity was quantified and stability during dispatch and storage were assessed in accordance with ISO Guide 35:2006. The certified value was obtained from the gravimetric preparation of mixtures, taking into account the purity of the base materials. The values were confirmed with two datasets obtained using the reference measurement procedure for HbA1c [ , , ]. Uncertainties of the certified values were calculated in accordance with the Guide to the Expression of Uncertainty in Measurement (GUM) and include uncertainties related to possible inhomogeneity, instability and characterisation. The material is intended for the calibration of the IFCC reference measurement procedure or for the assessment of method performance. As with any reference material, it can be used for establishing control charts or validation studies. The CRM was accepted as European Reference Material (ERM®) after peer evaluation by the partners of the European Reference Materials consortium.JRC.D.2-Standards for Innovation and sustainable Developmen

Development and Preparation of a New Serum Protein Reference Material: Feasibility Studies and Processing

Background: The availability of matrix reference materials is essential for the standardisation of (immuno)assays used to measure proteins. The reference material ERM-DA470 (previously called CRM470) certified in 1993 has led to a large degree of harmonisation of these assays. A new serum protein reference material has now been produced (ERMDA470k). It is intended to replace ERM-DA470, and will additionally be certified for beta2-microglobulin (B2M). Methods: Serum from 390 healthy donors was pooled and processed so as to stabilise, delipidate and 'maturate' it. Purified C-reactive protein (CRP) and recombinant B2M were added. Pilot batches were produced to study the stability, homogeneity, and commutability of the material. On the basis of the results with the trial batches it was decided to proceed with the processing of the main batch of a candidate reference material. Results: Two pilot batches were produced and the processed and spiked serum lyophilised after filling (1 mL). The B2M in the material was shown to be stable and commutable. For CRP, it was discovered that freeze-drying led to a decrease in measurable protein. The main batch of candidate reference material was produced and fulfilled the required criteria in terms of optical transparency, homogeneity and stability. Conclusions: A new serum protein reference material has been produced with the properties required for a serum protein reference material for 14 proteins. An apparent loss of CRP of approximately 20% was observed upon freeze-drying of the material.JRC.D.2-Reference material