Newsletter Food Allergen Community

AOAC Food Allergen Community

NEWSLETTER Volume 7 | Issue 3

IN THIS ISSUE

2016

Editorial Comment

Editorial Comment

Christmas time - and some presents have already been delivered during the year

Christmas time - and some presents have already been delivered during the year

1

Featured Article

2

Fostering Consistent Guidance on Food Allergen Management Globally

2

News

4

Allergen analysis in support of consumer protection, quality control and regulatory enforcement

4

1st International Conference on Food Analysis (ICFA)

5

Scientific Developments

6

Allergen detection using mass spectrometry: MRM3 as a novel approach for increased specificity and sensitivity

AOAC also called a thought leaders meting to start a new ISPAM activity for food allergens, including ELISA and PCR methods, but excluding mass spectrometry methods. During a call at eh end of November, a draft SMPR for ELISA methods was discussed.

6

Major challenges for reliable detection of allergens in food products

7

Will allergies soon be curable?

8

During the annual meeting in Dallas, the two allergen sessions, both discussing LC-MS/MS based approaches and reference materials were among the best attended sessions (you will find some more details in this edition). And we’re very happy to report that a young scientist from Belgium, Melanie Planque, won the poster award and delivered an excellent presentation.

Upcoming Events

8

Symposium Food Fraud Prevention and Effective Food Allergen Management8

This year was special - better. A lot of progress has been made in the field of food allergens, and many new activities have started, some milestones achieved. Back in 2015, AOAC started to develop a Standard Method Performance Requirement (SMPR) document for the analysis of food allergens using mass spectrometry. After numerous discussions and revisions, AOAC SMPR® 2016.002 was approved and published in the Journal of AOAC 99(4). In addition, two LC-MS/MS methods for multiple allergen detection were submitted and discussed at the AOAC Annual Meeting. At the December meeting of the European Organization for Standardization (CEN), the status of the subgroup (CEN TC 275 WG 12 AHG 4), which deals with chromatographic methods (including LC-MS/MS) was changed from ‘dormant’ to ‘active’, indicating that there will also be some developments at European level.

In this issue, you will find some more details on the highlights from the 2016 Annual Meeting as well as some interesting articles about developments at Codex Alimentarius regarding threshold levels. You will also find some news from ‘Down Under’, where a research team aims to develop allergen vaccines and therapeutic agents. Enjoy the read and best wishes for a peaceful Christmas and a happy New Year from the Editorial Team. Bert Popping  Editorial Board Member

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Editorial Team

Editor in Chief & Graphic Design:

Carmen Diaz-Amigo 

Editorial Members: James Roberts , Bert Popping , Samuel Godefroy  Terry Koerner, Jupiter Yeung This newsletter is a public document and can be distributed. Partial reproduction is permitted with the proper indication of the author and its source. For questions contact us at [email protected].

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2016

Featured Article

Fostering Consistent Guidance on Food Allergen Management Globally

Over the last 20 years, several actions have been taken by various players with the aim to enhance the protection of food allergic consumers. While several measures targeting the improvement of ingredient labelling, with emphasis on allergenic ingredients, have been harmonized thanks to standards promulgated by the Codex Alimentarius Commission, far more needs to be done in relation to the use of precautionary statements. Allergen precautionary statements continue to be diverse, to be used under different conditions and are not necessarily supported by risk assessment. As a result, these labelling practices are not helpful to consumers. When one examines the history of food allergen management, it is easy to note that most regulatory measures that were developed at the national level took place after the Codex Alimentarius Commission known as Codex and acting as the international food standard setting body, developed its global standard on food allergen labelling in 1999. Several years were required, and expert advice was mobilized at the global level by the World Health Organization (WHO) and the Food and Agriculture Organization (FAO) of the United Nations to come-up with a set of criteria for the identification of food ingredients that have the potential to be allergenic. These ingredients were then covered by mandatory declaration on food labels. Several domestic food regulatory policies were then amended and the world witnessed the development of a number of allergen labelling regulations and legislations in Australia/ New-Zealand, Europe, the United States of America, Japan, Canada and a number of emerging economies in Latin America and Asia. All these requirements were based on the Codex Standard and ensured to prevent the omission of declaration of food ingredients known to be priority allergens, when these ingredients are deliberately added to the recipe of a prepackaged food. This did not however address all issues allergic consumers and their care givers are having with food labels. The propagation of use of allergen precautionary or advisory labeling in the form of “may contain” statements and other iterations is undermining consumers’ confidence and their trust in the reliability of information offered by food labels. While precautionary statements were initially meant to address situations of possible cross-contamination due to

an allergenic ingredient, which cannot be avoided under reasonable food processing conditions, there was no clear guidance as to how they should be applied in a manner that is risk based. Statements of different sorts are popping on food labels, with no regard to what they may mean in informing the choice of allergic consumers and their families. Others are being used with no particular justification, other than to “cover a potential liability”. The practice has even led to witnessing precautionary statements on food labels that cover the entire list of priority allergens and that are longer than the list of ingredients itself. No wonder that confusion reigns, not only for consumers, but also amongst the health professional community, whose guidance is sought in helping allergic consumers manage their avoidance of potentially offending foods. The scientific community acknowledged the problem and agreed that investing in the development of the scientific foundations for a risk-based approach to be applied would contribute to addressing this problem. A decade later, several studies resulted in the generation of thousands of data points related to thresholds for food allergen reactions. Risk assessment methodologies have also been discussed and are being adapted to food allergens as a food hazard. Allergen analytical methods have been developed and are being used to help validate sanitation practices and other quality control and management measures. We have even witnessed the development and adoption of allergen control programs by the food industry sector in an attempt to create “order and structure” as to how allergen advisory labeling should be used. While food allergen-related recalls continue to top the list of food recalls in North America, Europe and some parts of Australasia, there is limited to no leadership from domestic food regulatory jurisdictions to move forward with clear regulatory measures focusing on allergen precautionary statements. Such measures would be the cornerstone for a more predictable environment for industry and consumers on food allergen management. Learning from history to address this conundrum can be useful in this case. I would argue that like what happened in the 1990s and early 2000s, it is time for Codex and its parent organizations to act. Attention needs to be given to the development of global guidance related to allergen thresholds, allergen precautionary labeling and management of cross-contamination and adventitious presence of priority

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2016

allergens in food manufacturing. These global standards could then lead the way in driving change in domestic food regulatory requirements in this area, which could then use the legitimacy of the international process. The time has come for Codex to lead again on this issue. I realize that suggesting such an approach, can be met with cynicism, given how long some international food standards can take in the making. But Codex has also shown that when its membership wants, it can. The first Melamine standards that Codex embarked on developing in 2009, took only one year between decision to act and adoption of the standard, making it the fastest agreed-upon Codex standard to date. Opponents to this approach would also argue that there is currently no interest on the part of Codex to work in this area, nor would there be resources made available. My argument back is that momentum can indeed be created. The case can be made for Codex standards being needed, given the

discrepancies of food allergen management policies and their impacts on international food trade. The case can also be made that scientific data has been generated over the past decade or more and could be relied-upon for international expert groups to be convened by FAO and WHO, to guide Codex with the relevant scientific advice. Finally, and should there be a will to act, funding could be mobilized from various government organizations currently struggling to move forward with addressing this issue and who would benefit from pooling resources under the auspices of a collective initiative. It is possible for history to repeat itself. Twenty (20) years after the initiation of the process that led to allergen labeling measures as we know them influenced by Codex leadership, action leading to renewed international guidance on food allergen management is overdue. Samuel Godefroy  Université Laval

The Rapid Alert System for Food and Feed 2015 Report Download Report 

Download Report 

September 24 - 27, 2017 Marriott Atlanta Marquis Atlanta, Georgia, USA  More Information » www.aoac.org

Participate! Scientific Sessions dedicated to Food Allergens & Food Allergen Community Meeting

EDUCATE NETWORK COLLABORATE

131st AOAC Annual Meeting & Exposition

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2016

News

Allergen analysis in support of consumer protection, quality control and regulatory enforcement

This year, the AOAC Annual meeting felt different. Better. What was very noticeable was the strong engagement of all stakeholders in focal areas this year. Allergen analysis was one of them. Besides the partially controversial discussions about which information is needed to accept mass spectrometry based methods as First Action Method, AOAC also launched an initiative to establish an ISPAM working group for immunological and molecular biological analysis of food allergens. The allergen sessions were extremely well attended, and the session organized by Romer Labs and co-chaired by Dr. Carmen Diaz-Amigo and Prof. Sabine Baumgartner had the highest attendance of all sessions during this years’ AOAC Annual Meeting. The session featured three parts: the first part reported about advances in reference materials for food allergens, reported by Prof. Roland Poms (MoniQA). Prof. Poms even showed some sachets of reference materials. In the second advances of LCMS/MS analysis of food allergens were discussed, presented by Prof. Sabine Baumgartner, Prof. Melanie Downs and Dr. Bert Popping. The third, not scheduled part, was a wider discussion with method and equipment providers, which were asked on stage, moderated by the previously mentioned presenters. As an introduction to this discussion, the evolution and lively dynamics of European directives and regulations for food allergen labelling were shown, followed by a comparison of results in proficiency tests (PT). It became clear that quality control and enforcement of food allergens were currently challenged by the differences in results between methods. It was mentioned that contrary to other PT where the assigned value for the sample is reported at the conclusion of the test, for food allergen analysis, assigned values are per applied method (e.g. per ELISA kit used by the participants of the PT). This is mainly due to the variability of results between the different methods used. Dr Håkan Emteborg from the EC JRC Institute for Reference Materials and Measurement (IRMM) explained that this is a ‘catch 22’ situation. Since there is no reference method, no certified reference material can be manufactured. And since there is no certified reference material, it cannot be

Panellists of the allergen session at the AOAC Annual Meeting. From left to right, first row: Scott Radcliffe (Romer Labs), Vincent Paez (Sciex), Jerry Zweigenbaum (Agilent), Steve Gendel (IEH Labs), Markus Lacorn (R-Biopharm), Tony Lupo (Neogen), Bert Popping. Back row: Melanie Dawns (University of Nebraska), Sabine Baumgartner (BOKU University)

established if a method is sufficiently reliable and robust to become a reference method. The discussion continued to the calibrants used for the individual methods. It became clear that currently all method providers - in the absence of certified reference materials - use their own material, which introduced variability. All method and equipment providers agreed that commutable reference material – if used as calibrant and quality control material, as it is currently in production and distribution by MoniQA, would help eliminate some of the variabilities between the methods and contribute to the harmonisation of results, ultimately benefiting consumers, food manufacturers, enforcement authorities and ultimately affected consumers. The panellists agreed that the use of a common reference material would be a good step forward. Further contributions to this discussion were made by Covance and IEH. The participating companies, namely Agilent, Neogen, R-Biopharm, Romer Labs and SCIEX were applauded by the audience and thanked for their engaging discussion. Bert Popping  Editorial Board Member

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First International Conference on Food Analysis (ICFA) The 1st International Conference on Food Analysis (ICFA) and the workshops on proficiency testing and food allergens were held in Melbourne, Australia (21st - 24th November 2016) with the theme “Where food analysis meets industry and regulation”. The conference brought together the themes on food quality and food safety, strengthening the partnerships between food industry, food trade, food legislation and regulation, public health and the science community. Approximately 180 delegates consisting of research scientists, those involved in analytical sciences, and those associated with public health, environment and agriculture, regulation and trade participated for the event. There were over 25 international delegates representing Asia, Europe, North America and other parts of the world. The conference was organised by The National Measurement Institute – Australia, ACS Laboratories, National Testing Authorities Australia, James Cook University, Victoria University, Melbourne University and Queensland Health and Forensic and Scientific Services.

A significant emphasis was given to food allergens including plenary presentation by Prof. Andreas Lopata from James Cook University providing insight on the current research in food allergy. Other presenters included Dr. Martina Koeberl (National Measurement Institute, Australia), on identification of common allergenic legume protein by mass spectrometry, as well as Adrian Rodgers and Dr. Alice Lee. The topics discussed in the workshop on food allergens included, new technologies for allergen detection, accreditation and proficiency testing, low level gluten detection, lupin as possible mandatory allergen in Australia, allergen detection problems in processed food products and how to report analytical allergen detection results. It is expected that the conference to be held two yearly in Melbourne. Martina Koeberl National Measurement Institute, Australia

Participants of the 1st International Conference on Food Analysis.

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2016

Scientific Developments Allergen detection using mass spectrometry: MRM3 as a novel approach for increased specificity and sensitivity Food allergy is one of the major challenges in consumer protection in industrialized countries. It is estimated that 150 million people worldwide suffer from allergic reactions to food and that about 5% of the population in westernized countries is affected. The “may contain” labelling of potential allergen contaminations has been shown to provide only limited protection for allergic consumers, partially because of its extensive use. An alternative strategy would be the monitoring of allergen trace contaminations using sensitive and specific analytical tools on the basis of clinically defined allergen threshold doses. As the availability of these clinical data has strongly increased in recent years, the analytical surveillance of allergen contaminations is now within reach. There is however still a need for the development of sensitive and quantitative analytical methods. Targeted proteomics using mass spectrometry is a promising alternative technique that offers increased specificity and reproducibility compared to currently used methods. As the available clinical data indicate that the detection of minute trace levels of food allergens is required for efficient consumer protection, highest sensitivity is needed for the MS-based detection. We therefore employed a bottom-up proteomics approach for the identification of allergen-specific proteotypic peptides from six different nut species (peanut, hazelnut, walnut, pistachio, almond, cashew). Following thorough experimental validation of specificity and sensitivity, 44 suitable tryptic marker peptides for the six nut species were identified. Figure 1. Principle of the MRM3 experiment.

To ensure highest sensitivity, a targeted proteomics method using triple-quadrupole instrumentation was developed. In addition, the analytical performance of multiple reaction monitoring (MRM) and MRM3, available on QTrap instruments, was compared. In MRM3-experiments, the precursor ion is isolated in Q1, fragmented in the collision cell and the fragment ion of interest is accumulated in the linear ion trap (Q3) and subjected to fragmentation, resulting in a secondary MRM transition (Fig. 1). For the targeted method development 38 MRM3 experiments specific to 18 proteotypic peptides (three per nut species) were optimized and in parallel respective MRM transitions were recorded for all peptides. All nut species were spiked at different concentrations ranging from 0.3 µg/g to 100µg/g in three different matrices (bread, ice cream, muesli). Limits of detection of the MRM3 approach were approximately 1 μg/g or below in fortified matrix samples. Compared to the MRM-based detection, the MRM3 method showed increase in sensitivity of up to 30-fold. Regression analysis demonstrated high linearity of the MRM3 signal in spiked matrix samples together with robust intersample reproducibility, confirming that the method is highly applicable for quantitative purposes. Notably, matrix interferences observed for some peptides in MRM-mode were not present in MRM3 data demonstrating increased specificity. To the best of our knowledge, this is the first study that systematically compares MRM3 with MRM for the analysis of complex foods. Jens Brockmeyer | University of Stuttgart Reference: Korte R, Brockmeyer J. (2016) MRM3based LC-MS multi-method for the detection and quantification of nut allergens. Anal Bioanal Chem, 408(27): 7845-7855. Abstract 

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Major challenges for reliable detection of allergens in food products Food allergies have increased impressively over the last few decades. In order to protect customers with food allergies, allergens must appear clearly on food labels. Labelling food products adequately can be hard, however, because of potential cross-contaminations during food production. To help food producers in this tedious task, the food community has developed analytical methods for food allergen detection. Nonetheless, there remain issues that must be addressed in order to harmonize food allergen control. Recently, although the number of food allergen analyses has increased dramatically, regulatory thresholds still have to be established. Systems proposing thresholds in food products (VITAL, EAACI…) have been developed, but these thresholds have not yet acquired regulatory force. Method sensitivities are mostly determined by spiking solvents or matrix extracts with allergens. This approach makes methods attractive, but creates a significant gap between declared and real thresholds, especially for heatprocessed food products. Some laboratories, aware of this shortcoming, have produced and commercialized incurred materials (FAPAS, MoniQA, LGC…). Another problem is that sensitivity determinations based on measurements in only one matrix cannot guarantee sensitive detection of allergens in processed matrices or in ones with a high fat content or high acidity. That a method is fit for purpose should thus be established carefully in each case.

Despite the emergence of incurred and standard materials (from producers such as the National Institute of Standards Technology and LGC Standards), there is a lack of standards for most allergens. Unfortunately, this leads to the use of different materials (walnuts, roasted walnuts, walnut milk…). Furthermore, different members of the food community report their results in different units: ingredients, soluble proteins (BCA quantification), total proteins (theoretical content), or protein (casein, ovalbumin…). The lack of standards and guidelines makes it very hard to compare methods, so that results are almost useless for laboratories and even worse for food producers. Concerned about this lack of harmonization, the AOAC Stakeholder Panel on Strategic Food Analytical Methods (SPSFAM) has recently established requirements for the evaluation of mass-spectrometry-based methods for allergens (SMPR 2016.002 “Standard method performance requirements for detection and quantification of selected food allergens”). In its guidelines, this panel recommends reference standards, target matrices, ranges of quantification, etc. The publication of validation guidelines and the development of reference standards and materials are first steps towards harmonization and method performance testing. Yet the food community still has a long way to go for full harmonization of methods. Melanie Planque | CER Group Nathalie Gillard | University of Namur

Solutions

Identified problems

Summary of the lack of harmonization encountered by the food community and possible solutions. Proteins Expression of results

Ingredients

Determination of method sensitivity Solution for harmonization

Soluble proteins (BCA quantification)

Total proteins (Theoretical content estimation)

Spiked Reference materials and standards Regulatory Thresholds

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Protein Casein αS1, β-lactoglobulin

Incurred

Regulatory thresholds

Guidelines Mass Spectrometry methods (SMPR 2016.002)

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Will allergies soon be curable? The team of professor Lopata from the Australian James Cook University is looking to develop vaccines or therapeutic agents for cure or lessen the effect of food allergies. The Queensland based group analyses the molecular structure of proteins with the aim to modify them so they can be used for treatment of allergies and/or diagnostic tools. Professor Lopata pointed out however, that these therapeutic agents would not be used for acute treatment of allergies like an epi-pen. The development is oriented more in the direction of desensitisation. At present, the group focus on fish. According to the working group, they have blood samples from more than 200 children who are allergic to different species of fish. Professor Lopata pointed out that there are already some diagnostic tools available for cod, but that exiting diagnostics to identify cod allergy are not fail-safe on one hand and that cod is not frequently consumed in Australia on the other hand. Their research will focus on fish species more frequently consumed in Australia. According to the group, fish diagnostic tools and therapeutic tools could be ready in 2 years and other foods for which allergies exists in four years. The research is funded by the National Health and Medical Research Council. James Roberts  Australian National Measurement Institute

Upcoming Events Symposium Food Fraud Prevention and Effective Food Allergen Management January 26-27, 2017 Bari, Italy Co-organised by MoniQA Association and CNR-ISPA

Professor Andreas Lopata (Left) and Dr Aya Taki (right). Source: www.abc.net.au

AOAC Food Allergen Community Newsletter Contribute with articles, news items or suggestions. Submission deadline for the 1st issue of 2017: March 24 Send your articles to [email protected] Topics for publication

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The AOAC Food Allergen Community is a forum serving the scientific community working on Food Allergens: The community aims to help AOAC INTERNATIONAL in its consensus-based scientific and advisory capacity on methods of analysis for allergens in foods and other commodities. It is also meant to serve the broader Stakeholder Community whose objectives it is to enhance the protection of food allergic consumers worldwide. Contact us at [email protected]

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Newsletter Food Allergen Community - focos

Newsletter Food Allergen Community - focos

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