Dr. Bert Popping

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1 ALLERGENS The Analytical Challenge to Meet Legislative Requirements and Consumer Demands Dr. Bert Popping

2 PART I INTRODUCTION TO FP6 PROGRAM MoniQA

3 Towards the harmonisation of analytical methods for monitoring food quality and safety in the food supply chain Towards the harmonisation of analytical methods for monitoring food quality and safety in the food supply chain Contract N0. FOOD-CT Contract N0. FOOD-CT

4 Funded by the European Commission within the Sixth Framework Programme (FP6) Topic T : Quality and safety control strategies for food (NOE) Coordinator: Roland Ernest Poms, ICC 33 member institutions from 20 countries / 4 continents Currently: over 100 registered institutions from 35 countries A total of 155 researchers including 40 doctoral students Currently more than 400 scientists worldwide Duration: 5 years starting 1 February 2007 EU funding: 12.3 M

5 Stakeholders ndustry / SMEs Food Manufacturers Retailer Food Industry Lobby Method providers / Test kit providers Providers of Reference Material Laboratories Trade organisations Authorities National / International Regulatory Bodies Validation, Standardisation, Proficiency Testing Bodies Authorities / Policy makers European Commission esearch & Science Universities Research institutes - PhD students - Young scientists - Senior Scientists Consumer Consumer Protection Group Consumer organisations The press and media

6 Working Groups 1. Microbiological Contaminants 2. Biotoxins/Phycotoxins 3. Chemical Contaminants 4. Food Allergens 5. Food Authenticity 6. Food Additives 7. Emerging Issues 8. Qualitative Method Validation Method Performance Quality Assurance and Control Sampling Measurement Uncertainty Legislation Socio-Economic Impact Traceability Harmonisation 9. Socio-Economic Impact

7 Working Groups Food Allergens Supporting databases Contributing to validation guidelines Method collection Validation studies certification Publishing position paper Consulting Training and dissemination / meetings Preparation for company audits Food Allergen Analysis Workshop, Halifax, CAN, May 2008 Method Performance Quality Assurance and Control Sampling Uncertainty Legislation Economic Impact Traceability Harmonisation

8

9 PART II ALLERGENS

10 LABELLING REGULATION IN EUROPE

11

12

13

14

15 and/or other allergens? like fish? mustard celery? regulated allergens or not??

16 2000/13/EC

17 Deals with: Product name Ingredient list BBE Irradiation storage conditions etc..but not: allergens

18 Amendment EC: Allergen Labelling Directive

19 Labelling Directive EC Mandatory labelling of ingredients containing or made of allergens:

20 Important: Labelling has to be understood by the consumer Labelling of allergens has to be understood by the (allergic) consumer: Maltodextrin -> Maltodextrin from wheat Lysozyme -> Lysozyme from egg Flavours -> Flavours from wheat

21 Problems Even highly processed products which no longer contain the offending substance (e.g. allergenic protein) need to be labelled No threshold is set for labelling of allergens, neither clinical, analytical nor action limit

22 emporary Exemptions from abelling Directive 2005/26/EC

23 Exemptions

24 Directive 2006/142/EC : Lupine & Molluscs

25 Summary of what we had so far 13/2000/EC : general food labelling directive 89/2003/EC : amendment to 13/2000/EC: Allergen labelling Directive 2005/26/EC: exemption from allergen labelling directive 2006/142/EC: added lupine and mollusc to labelling directive confused anybody???

26 Another Amendment in 2007

27 Finalising the Allergen and Exemptions List

28 The subtle differences to 2005/26/EC Which exemptions are no longer granted: Lysozyme from egg used in wines Albumin from egg used as fining agent Milk-casein used as fining agent in Applewine (Cidre) and wine Almonds and Walnut extracts used as flavours in Spirits Celeryoil Celeryseedoil. Celeryseedoleoresin Mustardoil Mustardseedoil, Mustardseedoleoresin Applications for exemptions can still be submitted at any time! => amendment of the amendment of the amendment of the.

29 but not quite yet final The Gluten Regulation 2009/41/EC JANUARY 2009

30 Gluten Labelling in 2009/41/EC Products as consumed by the final consumer: Gluten reduced products <= 100 mg/kg: shall bear very low gluten Gluten reduced products <= 20mg/kg: may bear glutenfree

31 DETECTION OPTIONS

32 Most Important Detection Methods USAGE TYPE ELISA PCR (LPA) LC-MS/MS

33 ELISA

34

35 ELISA

36 ELISA but Gluten ELISA Vendor A Vendor B Soy drink - Vanilla 21,86 ppm < MIN Soy drink - unsweet. 59,43 ppm < MIN Dry glucose < MIN < MIN Soy drink of another 15,53 ppm <MIN gluten-free Producer Soy flour < MIN < MIN FALSE-POSITIVES DUE TO MATRIX EFFECTS YOU NEED TO KNOW YOUR TEST & MATRIX!!

37 ELISA but Vendor A Simple handling 150 min

38 ELISA but Vendor A Problems with Std kurve

39 PCR

40

41 PCR Standard PCR / Real-time PCR LPA Technology

42 PCR but Indirect assay: detects only DNA, not the allergenic protein. Theoretically, DNA may be present but the offending allergenic protein may be absent Quantification of protein levels not possible (DNA- Quantification may not relate to proteins present) LPA: rarely used takes 16hrs

43 LC-MS/MS

44

45 LC-MS/MS but Sensitivity not as low as desireable Still laborious Very expensive equipment Quantification still a major issue

46 SUMMARY Three major techniques: ELISA, PCR/LPA, LC- MS/MS No perfect technique all techniques have their place

47 THANK YOU FOR YOUR ATTENTION Bert Popping

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