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Wednesday - November 3, 2021
9:00-13:00 Vendor webinars
Thursday - November 4, 2021
13:30-18:30

Virtual event highlighting current Trends & Views - RAFA 2021 : PROGRAM

Wednesday - November 3, 2021 Chairs: Jana Hajslova & Michel Nielen
Thursday - November 4, 2021 Chairs: Jana Pulkrabova & Stefan van Leeuwen

13:45-14:00

Welcome at RAFA 2021

Jana Hajslova & Michel Nielen, RAFA chairs
Session 1

Analytical challenges I
14:00-14:20

L1

Fighting food frauds exploiting chromatography-mass spectrometry & infrared technologies: scientific literature vs real industrial approaches

Fraud in food commodities is very common in all regions of the globe and today occurs at different places of the supply chain: the demand for rapid and confirmatory analytical methods has increased in recent years.

This presentation intends to illustrate a comparison between chromatography-mass spectrometry (mainly) & infrared technologies (few other examples) solutions to detect food fraud suggested in scientific literature and practices actually implemented by food companies.

This will be done taking into consideration several different types of matrixes/food chains (wheat/cereals, fruits, nuts and nut products, eggs & egg products,...) and integrating the information about industrial behaviour, directly collected through informal interviews and official open days & workshops; executed during FoodIntegrity European Funded Project.

The academic and food industry worlds are not aligned thus far and should interact better, in particular revolving around the flexibility and performances of chromatography-MS solutions.

References:
[1] Suman et al.
Fighting food frauds exploiting chromatography-mass spectrometry technologies: Scenario comparison between solutions in scientific literature and real approaches in place in industrial facilities Trends in Analytical Chemistry 142 (2021) - https://doi.org/10.1016/j.trac.2021.116305
[2] McGrath et al.
The potential of handheld near infrared spectroscopy to detect food adulteration: Results of a global, multi-instrument inter-laboratory study Food Chemistry 353 (2021) - https://doi.org/10.1016/j.foodchem.2020.128718
[3] De Girolamo et al.
Tracing the Geographical Origin of Durum Wheat by FT-NIR Spectroscopy Foods 8 (2019) - https://doi.org/10.3390/foods8100450
[4] Suman et al.
Hazelnut and apple products traceability through Near Infrared spectroscopy approach Proceedings of Sensor Fint 2021 - Smart Spectral Sensors for Agri-Food Quality and Process Control Workshop - Porto (Portugal) 30/09/21-01/10/21

Michele Suman
Barilla G. R. F.lli SpA, Advanced Laboratory Research, Parma, Italy
14:20-14:40

L2

How can we measure and tackle the world wide occurrence of mycotoxins in view of climate change?

The Food and Agriculture Organization (FAO) estimated the global food crop contamination with mycotoxins to be 25%. In order to assess the rationale for this figure which dates back to prior 1985, the relevant literature was reviewed and data of around 500 000 analyses from the European Food Safety Authority and large global survey for aflatoxins, fumonisins, deoxynivalenol, T-2 and HT-2 toxins, zearalenone and ochratoxin A in cereals and nuts were examined by M. Eskola et al. 2019. Using different thresholds, i.e., limit of detection, the lower and upper regulatory limits of European Union (EU) legislation and Codex Alimentarius standards, the mycotoxin occurrence was estimated. Impact of different aspects on uncertainty of the occurrence estimates presented in literature and related to our results are critically discussed. Current mycotoxin occurrence above the EU and Codex limits appears to confirm the FAO 25% estimate, while this figure greatly underestimates the occurrence above the detectable levels (up to 60-80%). The high occurrence is likely explained by a combination of the improved sensitivity of analytical methods and impact of climate change. The latter leading to unexpected occurrence patterns and a shift of pathogens towards the poles. It is of immense importance that the detectable levels are not overlooked as through diets, humans are exposed to mycotoxin mixtures which can induce combined adverse health effects. Hence, both, sensitive and fully validated LC-MS-based methods as well efficient integrated strategies to mitigate the mycotoxin issue along food and feed chains are needed. This presentation will summarize the state-of-the-art and recent developments in tackling these global challenges.

Rudolf Krska
University of Natural Resources and Life Sciences, Vienna, IFA-Tulln, Austria
14:40-15:00

L3

Potential of imaging mass spectrometry in characterisation of food crops

Mass Spectrometry Imaging (MSI) is a technique used to visualize the spatial distribution of biomolecules such as peptides, proteins, lipids or other organic compounds by their molecular masses.
By collecting mass spectra spot by spot, the sample is scanned and the MS data is used to map the distribution of selected compounds across the sample. This results in pictures of the spatially resolved distribution of a compound pixel by pixel. Most common ionization technologies in the field of MSI are DESI imaging and MALDI imaging. This approach has been used so far mainly in to study the spatial distribution of biomolecules within tissues, and other biological media.
Sample ionization is a major challenge of the technique, especially when the target is on small molecules. In addition, data mining is often cumbersome, due to the size of the generated dataset. Therefore, dedicated, and user-friendly software may be of great support for interpretation.
Although often applied in clinical science, MSI can be of great support in understanding the effect of biotic and abiotic stressors in crops. Plant response to environmental factors may not always end up in an over/under accumulation of specific metabolites, but it more often gives rise to change in their spatial distribution.
We have recently applied MSI for elucidating the spatial distribution of secondary metabolites in plants, following mycotoxin contamination. Atmospheric-pressure scanning microprobe matrix-assisted laser desorption/ionization (AP-SMALDI) MSI, for instance, is versatile enough to cover a broad range of plant metabolites, allowing simultaneous localization clues about the putative activation and the spatial, histological distribution of specific metabolic pathways. In this regard, modern metabolomics coupled with MSI may allow a proper insight into the interplay between plants and mycotoxins, highlighting potential variations and correlations between multiple metabolites.
In this talk, the opportunities and the limitation offered by MSI will be discussed, with a particular focus on the mapping of small molecules in crops.

Acknowledgements: LR and CDA are grateful to Prof. Bernhard Spengler and Dr. Dhaka Ram Bhandari, Institute of Inorganic and Analytical Chemistry, Justus Liebig University Giessen (Germany) for the scientific and technical support in the MSI analysis as well as for the fruitful collaboration and discussion. LR acknowledges the German-Italian Research Short-Term Scholarship (DAAD-2019, ID:91714320) funded by the German Academic Exchange Service (DAAD).

Chiara Dall'Asta
University of Parma, Parma, Italy
15:00-15:20

L4

Dimensional Chromatography with Quadruple Parallel Mass Spectrometry, LC3MS4, for Infant/Adult Formula Analysis

Two-dimensional liquid chromatography (2D-LC) is becoming less exotic and more mainstream as commercially available instruments proliferate. Despite the impressive capabilities of 2D-LC, there are times when even 2D-LC is not sufficient for an adequate separation. One-dimensional reversed-phase HPLC is not adequate to completely resolve all milk (or infant formula) triacylglycerol (TAG) molecular species, which are highly complex, and contain many isomers of TAGs, especially TAGs with short-chain fatty acids. Even conventional 2D-LC is not completely satisfactory. A new approach was developed, in which two second dimensions were used simultaneously for dual parallel comprehensive 2D-LC, giving three dimensions of chromatography, with quadruple parallel (x4) mass spectrometry. A pair of contact-closure (CC) controlled UHPLC switching valves were coupled to a timed CC circuit to allow the second 2D separation, ²D(2). The first second dimension, ²D(1), used the commercially available system, in which conventional 2D-LC improved the separation of components, and all components eluted in the first modulation period from a 50 mm C30 column. The 2D(2), which employed a 100 mm C30 column, produced an even better separation of components, but required more than one modulation period to elute, resulting in multi-cycle chromatography or "controlled wraparound" chromatography. Electrospray ionization (ESI) mass spectrometry (MS) was used in parallel with atmospheric pressure photoionization (APPI) MS for LC1MS2 to monitor the first dimension RP-HPLC, while ESI-MS monitored the 50 mm C30 UHPLC (LC1MS1) and ESI-MS also monitored UHPLC using the 100 mm C30 column (LC1MS1), to produce LC1MS2 x (LC1MS1 + LC1MS1) = LC3MS4.

Craig Byrdwell
U.S. Dept. of Agriculture, Agricultural Research Service, Beltsville Human Nutrition Research Center, USA
15:20-15:30

L5

Towards decentralized food safety: a FoodSmartphone perspective

Until recently, food safety was considered a job for governments and industry. However, as consumers become more aware about food safety and the associated risks, we are witnessing a rise in user-friendly food safety tests, such as lateral flow immunoassays (LFIAs) which will allow for affordable and sensitive on-the-go food testing - that's where FoodSmartphone comes in. We believe that smartphones have the potential to transform food analysis, allowing consumers to test foods in their kitchen and producers to test crops in the field (step 1). The FoodSmartphone approach can also reduce laboratory costs, as high-end analytical instruments, such as mass spectrometry (MS), will be reserved for suspect samples which have been initially screened by end-users with a smartphone-based method (step 2). All that is needed is a portable assay able to detect a target compound or a specific class of compounds, a smartphone and, in some cases, lightweight 3D-printed, auxiliary parts for optimised image capture. This talk will give an overview of the FoodSmartphone project, touching on a step 1 integrated device for LFIA based total allergen detection by end-users and an innovative step 2 method for direct analysis of LFIAs for mycotoxin detection by MS. We envision that the concepts and techniques presented here will pave the way for the FoodSmartphone approach by combining on-site screening and lab-based confirmation of a range of food safety applications.

Georgina Ross
Wageningen University & Research, Wageningen, The Netherlands
15:30-15:40

Chat discussion with speakers L1-L5

Led by Jana Hajslova & Michel Nielen
15:40-16:00

Refreshment break
Session 2

Metabolomics
16:00-16:20

L6

Metabolomics and lipidomics approaches to advance food Science and nutrition research

The main objective of this presentation is to present the latest results obtained in our research group (Foodomics Lab) regarding the use of metabolomics and lipidomics techniques based on LC-HRMS and GC-HRMS to investigate the neuroprotective activity of bioactive compounds obtained from different natural sources (including algae, plants, by-products of the olive oil and orange juice industry, etc.) combining green extraction processes, a systematic study of the chemical composition using advanced analytical techniques (GC-QTOF-MS/MS and LC-QTOF- MS/MS), with in vitro models and cellular models of Alzheimer's disease. The conclusion of this work is that metabolomics and lipidomics techniques help to identify biomarkers related to the neuroprotective activity of the most active natural compounds found in model cells.

Alejandro Cifuentes
National Research Council of Spain, Madrid, Spain
16:20-16:40

L7

Deciphering the gut microbiota function by LC/GC-MS metabolomics approaches

Dietary patterns, or the food we eat, are the sum of many small molecules foreign to the body. After being ingested and digested, nutrients are altered by the trillions of microorganisms that inhabit our gastrointestinal (GI) tract, shaping the chemical structures of such compounds and thus modifying the lifespan, bioavailability, and biological effects. In this context, dietary patterns modulate the gut microbiome (community) and alter its functions by modulating the production of different gut microbial metabolites (GMMs). To date, an array of in vitro and in vivo models have been proposed to study gut microbiota functionality. In most cases, 16S rRNA has been predominantly used to define which microorganisms are present. On a few occasions, metagenomics has also been employed to resolve differences in the functional potential of microbial communities. However, metagenomics alone offers little information on which microbial traits contributed to human physiology. Since genes predicted from metagenomes may not necessarily be expressed. For this reason, the combination of multi-omics data revealing metabolic activities is highly required to decipher diet-gut microbiota interactions and the intestinal ecosystem.
During this presentation, I will discuss my vision of the diet-gut microbiota field and what we can achieve in the future. Preliminary data will accompany this presentation. First, the function of two distinct gut microbiota compositions, from obese and lean healthy subjects, fed with different foods (apple, apple pectin, and cellulose) will be compared. In this project, the metagenomic analysis revealed differences at the taxonomical level, and then the microbiome's functional activity was investigated in-depth by different pipelines. A further goal of this project is to measure metabolic outcomes of lean and obese gut microbiotas. Because metabolomics directly measures metabolites, these end-products may act as signaling markers at the local level (gut epithelium-gut microbiota) and promote gut health. We paid particular attention to the biotransformation of dietary fibers and phytochemicals, such as proanthocyanidins. Lastly, we investigated gut epithelium responses to specific gut microbial metabolites.

Josep Rubert
Wageningen University & Research, Wageningen, The Netherlands
16:40-17:00

L8

Confidence in metabolite annotations from high-resolution MS/MS in food digestion along the human GI tract

The human small intestine remains an elusive organ to study due to the difficulty of retrieving samples in a non-invasive manner. Stool samples as a surrogate do not reflect events in the upper gut intestinal tract. As proof of concept, this study investigates time-series samples collected from the upper gastrointestinal tract of a single healthy subject. Samples were retrieved using a small diameter tube that collected samples in the stomach and duodenum as the tube progressed to the jejunum, and then remained positioned in the jejunum during the final 8.5 hours of the testing period. Lipidomics and metabolomics liquid chromatography tandem mass spectrometry (LC-MS/MS) assays were employed to annotate 828 unique metabolites using accurate mass with retention time and/or tandem MS library matches.
However, False Discovery Rates (FDR) for LC-MS/MS annotations remain poorly studied. Which adduct species should we expect? At which (relative) retention time should a compound be found? How many alternative isomers and isobar molecules should we consider? We work on integrated workflows using 19 published databases and software packages to assist in structural characterizations, including MassBank.us with more than 650,000 public MS/MS spectra. Here, we show how these packages were employed, with extensions for (1) NIST hybrid search to yield chemical class information on all unknown compounds that did not have direct hits in experimental or in-silico MS/MS libraries; (2) "Entropy Similarity" as measure for MS/MS matching to improveo FDR over classic dot-product similarity matching ; (3) retention time libraries for both HILIC and RP liquid chromatography methods for more than 4,000 metabolite standards, used for retip.app retention time prediction; (4) hydrogen/deuterium exchange data to limit the chemical search space by the number of acidic protons in MS/MS spectra.
Annotated metabolites were clustered based on correlation to reveal sets of biologically related metabolites. Typical clusters included bile metabolites, food metabolites, protein breakdown products, and endogenous lipids. Acylcarnitines and phospholipids were clustered with known human bile components supporting their presence in human bile, in addition to novel human bile compounds 4-hydroxyhippuric acid, N-acetylglucosaminoasparagine and 3-methoxy-4-hydroxyphenylglycol sulfate. Food metabolites were observed passing through the small intestine after meals. Acetaminophen and its human phase II metabolism products appeared for hours after the initial drug treatment, due to excretion back into the gastrointestinal tract after initial absorption. This exploratory study revealed novel trends in timing and chemical composition of the human jejunum under standard living conditions.

Oliver Fiehn
UC Davis West Coast Metabolomics Center, Davis, USA
17:00-17:20

L9

One class modelling, a simple approach to botanical authentication

One-class modelling is a non-targeted approach based on principal component analysis (PCA) for authentication of foods or botanical materials. Neither an adulterant nor a specific component (marker compound) needs to be specified. A one-class non-targeted method employs only the data for the samples of the authentic material and any other material is potential adulterant. Non-targeted data are generally comprehensive chromatographic or spectral data that encompasses as many of the samples chemical properties as possible. For example, flow injection mass spectrometry will generally yield a nominal mass spectrum (profile) of around 1000 ions (e.g., m/z 100 to 1000 m/z) or variables. Since neither an adulterant or marker compound is specified, adulteration can only be detected if one or more of the variables of the unknown sample are statistically different from that of the authentic profile. Thus, the emphasis has shifted from what is the adulterant to simply examining deviations in the variables.
PCA provides two statistical values that express the fit of a sample to the model. The Hotelling T² statistic provides the sample variance within the model and the Q statistic that provides the distance of the sample from the model. The latter is the best means for determining subtle variations between a sample and the model. In addition, pre-processing with autoscaling (division of all the data for each variable by the standard deviation of the variable) provides normalization of the variance of each variable to a value of 1.0. This reduces the impact of variables with large intensities and places equal emphasis on all variables. Thus, the Q statistic for PCA with autoscaling is very sensitive to differences in intensities. This sensitivity is increased if the intensity of the model variable is small. Thus, adulteration, which presents a new signal at a variable that was previously at the baseline level, is readily observable. Adulteration based on an increase in an existing peak is less easily detected in terms of absolute intensity but equally detectable in terms of relative intensity change.
The relative sensitivity of the Q statistic for any variable can be easily predicted. In silico modelling can be used to predict the sensitivity of any variable.
With the pure spectra of any adulterant, in silico modelling can predict the limit of detection. Thus, one-class modelling using principal component analysis with autoscaling pre-processing and analysis of variance using the Q statistic is a powerful tool for detecting adulteration and predicting limits of detection.

James Harnly
U.S. Dept. of Agriculture, Agricultural Research Service, Beltsville Human Nutrition Research Center, USA
17:20-17:40

Chat discussion with speakers L6-L9

Led by Jana Hajslova & Michel Nielen
17:40-18:30

Opportunities for networking

Session 3

Food fraud & authentication
09:00-09:20

L10

From targeted to non-targeted analysis for food authentication: challenges of change

It is scientifically agreed that non-targeted analytical methods for food and feed authentication have the potential to strengthen official control. In particular, because these methods are able to uncover unknown adulterations in addition to the detection of already known fraudulent practices. The investigation of the complete spectrum or chromatogram of a product sample in question in comparison to appropriate reference data allows its comprehensive characterisation (like a fingerprint) and enables the detection of any deviation from typical product properties.
However, the ongoing process to implement non-targeted analysis alongside targeted analysis is accompanied by various challenges throughout the entire analytical process, from sampling to data handling. From our perspective, three aspects need further attention and will be taken into account in this presentation:
A) Adulteration: Development of innovative approaches for the detection of unknown additions/anomalies,
B) Standardization & Quality Assurance: Efforts with regard to comparability and validation of fingerprinting methods, and
C) Digitalization: Creation of user-friendly solutions for data management, exchange & evaluation.

Carsten Fauhl-Hassek
Federal Institute for Risk Assessment, Berlin, Germany
09:20-9:40

L11

Diving into the beer metabolome and discover signatures of materials, technologies and reactions

The last years have seen a tremendous development all fields within food analysis and the generic field of foodomics [1]. In particular for food metabolomics, the number of identified and quantified metabolites has dramatically increased [2]. Therefore, metabolomics is a promising concept to unravel the complex metabolome of beers.
In this study, the compositional space of a diverse set of beer samples is uncovered by a single analytical approach using direct injection Fourier Transform Ion Cyclotron Mass Spectrometry (FTICR MS) for the first time. The unrivaled resolution on mass measurements combined with mass accuracy and compositional networks uncovers and assigns structural information to thousands of yet unknown metabolites in the beer matrix [3]. Multiple statistical models enable the assignment of different molecular fingerprints to certain beer attributes such as the beer type, the grain used, the use of specialty dark malts or the type of fermentation. The annotation of sum formulae provides information in excess of the m/z space and reveals the nature of discriminating features as beers from barley, wheat, rice and corn can be differentiated from each other by sets of discriminating markers [4]. For instance, benzoxazinoid hexosides deriving from the wheat's secondary metabolism are uncovered as significant markers for the use of whole wheat grains, which raises the potential of FTICR MS for quality control and inspection purposes. The detection of fingerprints for light versus dark and top versus bottom fermented beers reinforces the role of FTICR MS in fundamental studies revealing Maillard reaction patterns [5].

References:
[1] Rychlik M, Kanawati B, Schmitt-Kopplin P (2017) Foodomics as a Promising Tool to Investigate the Mycobolome, Trends in Analytical Chemistry. doi.org:10.1016/j.trac.2017.05.006
[2] Rychlik M, Schmitt-Kopplin (2020) Reading from the Crystal Ball: The Laws of Moore and Kurzweil Applied to Mass Spectrometry in Food Analysis, Front. Nutr. 7:9. doi: 10.3389/fnut.2020.00009
[3] Pieczonka S, Lucio M, Rychlik M, Schmitt-Kopplin P (2020) Decomposing the molecular complexity of brewing, npj Science of Food (2020) 4:11. https://doi.org/10.1038/s41538-020-00070-3
[4] Pieczonka S, Paravicini S, Rychlik M, Schmitt-Kopplin P (2021) On the Trail of the German Purity Law, Front. Chem. 9:715372. doi: 10.3389/fchem.2021.715372
[5] Pieczonka S, Hemmler D, Moritz F, Lucio M, Zarnkow M, Jacob F, Rychlik M, Schmitt-Kopplin P (2021) Hidden in its color: A molecular-level analysis of the beer's Maillard reaction network, Food Chem. 361: 130112. doi: 10.1016/j.foodchem.2021.130112

Michael Rychlik
Technical University of Munich, Freising, Germany
09:40-10:00

L12

Alternative proteins for conventional animal products in China: regulatory process

The globe demand for protein is changing, creating opportunities in the agricultural and food market for several alternative sources of protein s such as cell-based meat and plant-based meat, as well as fermentation protein bases meat. The presentation will address the range of alternative proteins from perspective of science, food safety, technology and regulatory aspects. It offers an opportunity to discuss the novel technology of plant-base and cultivated meat. With the proliferation of new alternative proteins, food safety authorities need to be prepared to regulate new food categories. Understanding the technology and the process behind the alternative proteins production is essential to create regulation that will protect the safety of consumers and will allow them to make choices.

Yongning Wu
China National Center for Food Safety Risk Assessment, Beijing, China
10:00-10:10

L13

Wine varietal identification: solution of an uneasy task

One of the major challenges in wine analysis [1] is authentication of grape variety from which the wine is produced. Here, we have developed and validated an authentication method based on metabolomic fingerprinting, using ultra-high-performance liquid chromatography coupled with quadrupole orbitrap high resolution mass spectrometry (U-HPLC-HRMS/MS). In total, 101 red and 97 white wines, five varieties in each group (all authentic), were analysed within our study. Filtered samples, without any other processing, were directly injected into system. After the data mining and data pre-treatment steps, principal component analysis (PCA) was used to explore the data structure. The obtained unsupervised PCA models revealed a notable clustering according to the grape varieties. Subsequently, orthogonal partial least squares discriminant analysis (OPLS-DA) was used to create supervised binary models. To obtain the best models possible, an in-house R script for optimization of data filtration parameters was developed and implemented. From over 85 000 models calculated, those with the highest prediction abilities were chosen, validated, and arranged into a Rooted Binary Decision Directed Acyclic Graph (RBDDAG), which was used for wine variety authentication. In case of white wines, 96 % of samples in positive (ESI+) and 94 % of samples in negative ionization mode (ESI-) were correctly classified. Regarding red wines, 95 % of samples in ESI+ and 94 % of samples in ESI- were correctly classified. Worth mentioning that the aforementioned accuracies are calculated for the whole RBDDAG structure. Multiple individual binary models achieved prediction ability as high as 100%, however, for some of the binary models present in the RBDDAG structure, correct classification of the grape variety appeared to be more problematic due to a higher genetic (and therefore metabolomic) similarity between varieties. The results obtained within our study indicate the potential suitability of the developed method for official control purposes.

References:
[1] https://ec.europa.eu/jrc/en/research-topic/food-authenticity-and-quality

Leos Uttl
University of Chemistry and Technology Prague, Prague, Czech Republic
10:10-10:20

L14

The story of a leaf: a 2-tiered system for tea authenticity

Tea is the world's most popular beverage and have been traded worldwide for centuries, with an annual market worth $200 Bn (2019) with >10% growth rate. Its difference in geographic origin, quality, and taste leads to a huge variance in its price (With more than >50 folds for certain GI products). Recent studies and reports have also revealed the potential fraud issues in developing countries and tea extract drinks. The vulnerability of the global tea supply chain and its safeness have therefore become as major concerns for consumers and technical challenges towards regulation inspections. As an extension and forward program for EU-China collaboration schemes, we aim to develop a 2-Tiered analytical toolbox under the global network and platform of IGFS and integrate with artificial intelligence and machine learning to enhance its performance. Eventually to provide solutions for end users of deferent application levels.

Di Wu
Queen's University Belfast, Belfast, United Kingdom
10:20-10:40

L15

Food authenticity from a regulatory science perspective

The fight against food fraud has been explicitly mentioned in the European Green Deal as a policy priority ("... actions to combat food fraud, including strengthening enforcement and investigative capacity at EU level..."). International collaboration on food safety research is already well established, while for food authenticity this is not yet the case. Therefore, the fight against food fraud calls for an international and systemic approach involving cooperation and consultation among all stakeholders at all levels of the food chain. There is a great need for international harmonization in order to be able to detect and, what is more important, prevent or at least minimize fraud. To bridge this gap and support European Commission services regulating the feed-food chain as well as Member States authorities, the EC Knowledge Centre for Food Fraud and Quality was created and installed at the Joint Research Centre. The main remit of its operation is to provide validated knowledge for addressing the issue in a holistic manner, such as the design principles of early warning systems, horizon scans, vulnerability assessment tools and appropriate analytical methods, which are among the critical elements for equipping regulatory agencies as well as industry in the fight against food fraud.

Franz Ulberth
European Commission, Joint Research Centre, Geel, Belgium
10:40-11:00

Chat discussion with speakers L10-L15

Led by Jana Pulkrabova & Stefan van Leeuwen
11:00-11:20

Refreshment break
Session 4

Analytical challenges II
11:20-11:40

L16

Comparison of analytical methods and results: "Devil is in the details"

When comparing analytical methods, even small, seemingly insignificant details or parameters may have considerable impact on the reported results. Official or standard methods try to address these issues but their implementation might not be easy or practical if there are too prescriptive in terms of the employed equipment or products. Therefore, a balancing act is needed to allow the use of equivalent instruments or items, without significantly affecting the method performance and results. System suitability and data acceptability criteria are very important in that respect in addition to highlighting critical method steps, parameters or issues, such as potential interferences.
This presentation will explore various examples of details, method parameters and analytical approaches that could play an important role in the overall analysis and lead to differences in analytical results. For instance, many details should be considered in the initial preparation of the received samples, which is commonly a critical step in the overall process, yet often only marginally described in the actual methods. The use of mass spectrometry for analyte detection reduced the need for highly optimized chromatographic separations and opened door to faster analytical runs, but there are cases where chromatographic resolution of certain peaks (may not be only our target analytes) are important for accurate results even when mass spectrometric detection is used. And details behind calibration approaches or the way that the result is expressed as represent another set of examples of potential pitfalls that could be encountered in the analytical practice.

Katerina Mastovska
Eurofins Scientific, USA
11:40-12:00

L17

Scientific and regulatory challenges and developments for EU Reference Laboratories in food safety areas

After the food incidents of the 90's the European Commission adopted the General Food Law, GFL (Regulation (EC) No 178/2002) setting the general principles, requirements and procedures related to all stages of food and feed production and distribution. Many legal acts were regularly adopted, setting for example new maximum limits for substances in various food or feed commodities. While the GFL is still relevant twenty years later, it needs to address better new challenges and issues such as food sustainability, novel foods, nutrition labelling or recycling of food contact materials. How will this impact our analytical work? Concrete examples from several EURLs will be presented and discussed.

Piotr Robouch
European Commission, Joint Research Centre, Geel, Belgium
12:00-12:20

L18

Transition to a circular economy: analytical challenges for contaminants in the food chain

The circular economy rapidly develops in all parts of society, and the need to go circular is also expressed for food and feed production. At national and international level numerous initiatives towards the circular economy are established, both at the policy (e.g. EU's Green Deal), research and technology level. However, the food safety in a circular economy -crucial for the societal acceptance- does not receive the required attention yet. There are several examples where hazardous chemical substances show up in circular products, such as food and feed, but also in food contact materials like plastics. These substances may be present from previous applications, or generated during the recycling process. Clearly, there is a need to address the safety by state-of-the-art chemical analysis to unravel new contaminant sources, to determine exposure levels and support effective circular economy policies in the food chain. The aim of this talk is to demonstrate some examples of safety issues in the circular food production chains, and provide examples of testing strategies that can adequately address the needs for safety testing in these circular agrochains. Analytical challenges will be addressed and some examples will be provided where food safety is compromised by contaminants in circular food production.

Stefan van Leeuwen
Wageningen Food Safety Research (WFSR), part of Wageningen University & Research, The Netherlands
12:20-12:30

L19

Smartphone-based pesticide residue screening: goals achieved & challenges to be faced

The ever-increasing demand for food production, unfortunately, still requires a widespread use of pesticides to prevent pests and boost agricultural productivity. Among them, carbamate (CM) and organophosphate (OP) insecticides are widely applied due to their global availability as well as their minimum environmental persistence. Nevertheless, CMs and OPs show relatively high acute toxicity and their residues in food may result in chronic or even acute toxicity incidents. The toxicity of CMs and OPs arises from the inhibition of cholinesterases, a vital enzyme family in the neural system of insects and mammals including humans. To monitor pesticide residues, chromatographic methods are predominantly utilized. Although they provide selectivity, low limits of detection (LODs) and wide linear ranges, chromatographic methods are also costly and time-consuming requiring highly skilled personnel. To counter these limitations and provide an innovative solution as part of the EU-funded "FoodSmartphone" project, we have been working on a novel approach for CM and OP screening, which combined enzyme recognition to smartphones and 3D-printing technology.
Briefly, acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) assays with smartphone readout were developed and validated in various matrices including fruits, vegetables and human serum. In the first case, the assays can be used for rapid (within 10 min) OP and CM screening utilizing an in-house lab-on-a-chip (LOC) prototype device with certain autonomous features. In detail, (i) samples and reagents can be injected using silicone finger pumps eliminating the need of pipettes; (ii) acceptance/rejection of each measurement based on its dynamic response using a principal component analysis (PCA) of smartphone-generated data. The cost per prototype was estimated about 0.30 € and the attained detectability (LOD=0.050 mg kg-1 for carbofuran) was enough to monitor highly contaminated samples as those reported in the EU Rapid Rapid Alert System for Food and Feed (RASFF). Regarding the application in human serum samples, acute intoxication incidents due to neurotoxic OP insecticides have been occasionally reported. To enable rapid monitoring at an early intoxication stage, a microfluidic paper-based analytical device (µPAD) able to screen for chlorpyrifos-oxon, the toxic chlorpyrifos metabolite was developed. Impressively, the µPAD was pocket-size, cost-efficient (0.70 € per device), required minimal sample preparation (just a sample dilution) and attained robust results.
Undoubtedly the above-mentioned achievements significantly progressed the field, however, there are still challenges that need to be faced. Firstly, the use of multistep lab-based sample preparation protocols remains a bottleneck reducing the portability potential of smartphone-based methods. Another aspect that can significantly improve the current status is the development of smartphone apps for a specific assay providing real-time "one-click" results. Last but not least, large scale commercialization of smartphone-based methods will be achieved only if the result interphone variation will be adjusted. All in all, within this project, a paradigm shift towards portable food contaminant analysis has been initiated and at-home analysis can be realistically considered in the upcoming years.

Aristeidis Tsagkaris
University of Chemistry and Technology, Prague, Czech Republic
12:30-12:50

L20

Determining the source of our soya using an analytical toolbox

Food fraud is a growing and the problem for consumers, the food industry and regulators. Criminals will exploit any opportunity to gain economic advantages by cheating on the way that food is produced and false claims made about it. A new opportunity is emerging for fraudsters in terms of the strong desire of consumers to purchase sustainable food. This is turn has put pressure on the markets to deliver this. One of the most controversial crops harvested in the world is soya but to the strong links it has with sustainability and rainforest destruction. Many claims are now being made that soya being sold into the markets is sustainable yet data would show that only about 25% of the world's production is sustainably produced. A major research project is now underway to determine the origin of soya being sold into the markets in terms of the country and region of the country the soya was grown. In addition evidence of the soya being grown on recently deforested land will be sought. A wide range of analytical tools will be employed in the project provided as part of the Agilent Thought Leaders award made to the Institute for Global Food Security.

Christopher Elliott
Queen's University Belfast, Belfast, United Kingdom
12:50-13:10

Chat discussion with speakers L16-L20

Led by Jana Pulkrabova & Stefan van Leeuwen
13:10-13:20

Summary of RAFA 2021, Trends & Views

Michele Suman
Barilla Food Research Labs, Parma, Italy
13:20-13:30

Announcement of RAFA 2022

Jana Hajslova & Michel Nielen, RAFA chairs