Protection and Control of Geographical Indications - Tools for detecting IPR violations and food fraud
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1 Protection and Control of Geographical Indications - Tools for detecting IPR violations and food fraud Food Authenticity testing in Eurofins : the use of isotopic and NMR methods to control the origin and the traceability of food Freddy THOMAS 1st October 2015 EXPO Milano MK-PRE edt Version: 1
2 Eurofins laboratories: the origin Invention of the SNIF-NMR method (Site-specific Natural Isotopic Fractionation studied by deuterium Nuclear Magnetic Resonance) allowing the determination of the botanical origin of alcohol (Pr. G and M. Martin, Nantes University) 1987 Creation of EUROFINS Laboratory, specializing in the application of isotopic methods to food testing FINS is the French acronym for SNIF Today: Eurofins Analytics France laboratory based in Nantes (France) is the group Competence Centre for authenticity 2
3 Eurofins Analytics France Group authenticity competence centre Nantes, France 3
4 Authenticity Is a natural product or ingredient actually from its reputed source? Why is it necessary to check with analytical means? Verify raw materials Protect brand image Protect consumer s health and avoid headlines like. 4
5 Article UFC Que Choisir, 540, October
6 Eurofins Analytics France: more than 25 years of experience in food authenticity testing 1994 FAIM Food Authenticity: Issues and Methodologies European Project coordinated by Eurofins about available analytical solutions to check food authenticity 20 years later, after many EU projects (MEDEO, COFAWS, PURE JUICE, WINE DB, TRACE.)
7 Analytical tools for authenticity testing Chemical composition methods (e.g. chromatography) Identification & quantification of defined compounds Stable isotopes Verification of the origin of molecular species from the measurement of heavy/light stable isotope ratios Molecular Biology Identification of species, varieties, etc. through the DNA profile of the food product Profiling methods Investigation of the spectroscopic or chromatographic fingerprint of a whole food matrix 7
8 Stable isotopes: a powerful authentication tool The content of... Is influenced by And can reveal Examples 13 C, 2 H Botanical origin Addition of sugars from other plants Fruit juices 13 C, 2 H Synthetic pathway Addition of artificial substances Flavours : vanillin O, 2 H Origin of water Dilution with tap water Wines, NFC juices 13 C, 18 O, 15 N 2 H, etc. Geographic origin Mislabeling of origin All products 13 C, 15 N Diet Feeding regime Animal products 8
9 Stable isotope analyses Two main techniques for measuring stable isotope ratios SNIF-NMR Measures non-statistical deuterium distribution within a given molecule Method pioneered by Eurofins IRMS : Isotopic Ratio Mass Spectrometry Access to overall isotope ratios of a product or constituent Combustion or Pyrolysis of the organic product Can be coupled to GC unit 9
10 Official isotopic methods for food testing Method Product Fraction Technique Isotope ratios AOAC Official method fruit juice Ethanol (from fermentation) SNIF-NMR (D/H) I, (D/H) II,R AOAC Official method honey honey & proteins IRMS 13 C/ 12 C AOAC Official method maple syrup Ethanol (from fermentation) SNIF-NMR (D/H) I, (D/H) II,R AOAC Official method fruit juice & maple syrup Ethanol (from fermentation) IRMS 13 C/ 12 C AOAC Official method Vanillin Vanillin SNIF-NMR (D/H) i CEN (TC174 N108, ENV 12140) fruit juice Sugars IRMS 13 C/ 12 C CEN (TC174 N109, ENV 12141) fruit juice water IRMS 18 O/ 16 O OIV-MA-AS311-05, OIV/OENO 381/2009 wine & spirits Ethanol SNIF-NMR (D/H) I, (D/H) II,R OIV-MA-AS312-06, OIV/OENO 381/2009 wine & spirits Ethanol IRMS 13 C/ 12 C OIV-MA-AS2-12, OIV/OENO 381/2009 wine & spirits Water IRMS 18 O/ 16 O CEN, EN :2012 vinegar Acetic acid SNIF-NMR (D/H) CH3 CEN, EN :2012 vinegar Acetic acid IRMS 13 C/ 12 C CEN, EN :2012 vinegar Water IRMS 18 O/ 16 O AOAC = Association of Official Analytical Chemists (USA) CEN = European Committee for Standardization (EU) OIV = International Organisation of Vine and Wine 10
11 Authenticity of Wine and Spirits ORIGIN Agreement with declared botanical origin of alcohol Absence of unauthorised, exogenous ingredients Varietal compliance Stable isotope analyses Agreement with declared provenance Isotope ratios + other parameters 11
12 Authenticity of Wine and Spirits WINE COMPONENTS Water Ethanol IRMS Distillation SNIF-NMR 2 H/ 1 H 18 O/ 16 O 2 H/ 1 H CH 2 CH 3 OH IRMS 13 C/ 12 C 12
13 Official methods (AOAC) / limits (AIJN CoP) d 13 C ( ) AOAC Cane, maize Beet Orange (D/H) I ( ppm ) AOAC
14 Wine official control: EC 2729/2000 (isotopic database) ) Chaptalisation 2) Watering 3) Origin 4) Vintage From C. Guillou, BEVABS JRC Ispra, Italie 14
15 Wine official control: a challenge for Europe WINE Data Base project Grapes Project funded by the European Community under the Competitive and Sustainable Growth Programme ( ), Measurement and Testing Activity Establishing of a Wine Data Bank for Analytical Parameters for Wines from Third countries Contract N : G6RD-CT BfR, Germany (coordinator) FERA (ex CSL), United Kingdom Eurofins Scientific Analytics, France VUB, Belgium, BEVABS, JRC Ispra, Italy, UGOA, Italy, Technische Universität Berlin, Germany Universitatea de Stiinte Agricole si Medicina Veterinaria, Romania, National Institude for Wine Qualification, Hungary, Ministry of Finance, Czech Republic Micro-vinification acc. EC reg. 2729/2000 Reference wines Physico-chemical analyses Commercial wines SNIF-NMR ICP-MS HPLC, GC IRMS Chemistry Stable isotopes Elemental analysis «classical» analysis Statistics Within a country Between countries Exploratory analysis Discrimination analysis WINE 6 All commercial wines Model 1 (8 var.) Parameter = Distillation WATER Equilibration CO Combustion 2 ETHANOL Root O/ 16 O 13 C/ 12 C IRMS (D/H) I (D/H) II 2 H NMR Root 1 Hungary Czech Republic Romania South Africa 15
16 Applications to other products: traceability studies 2 H, 18 O Local climatic conditions 15 N, 34 S Agricultural practices, local «pollution» 13 C, 15 N Influence of diet in animals Confirmation of specifications for PDO, PGI labels Examples: Mozzarella, Extra Virgin Olive Oil, balsamic vinegar, Iberico ham. 16
17 Analytical tools for authenticity testing Chemical composition methods (e.g. chromatography) Identification & quantification of defined compounds Stable isotopes Verification of the origin of molecular species from the measurement of heavy/light stable isotope ratios Molecular Biology Identification of species, varieties, etc. through the DNA profile of the food product Profiling methods Investigation of the spectroscopic or chromatographic fingerprint of a whole food matrix 17
18 Need for new analytical approach : untargeted approach Official control today The future R&D project coordinated by Eurofins 18
19 High Resolution 1H Nuclear Magnetic Resonance Observation domain: H-containing compounds, ppm to % Key Strengths: Large linearity range Short Turn-Around Time High Reproducibility Discrimination power Lab 1 Lab 2 19
20 Benefits of a non-targeted method The 1 H-NMR spectrum is obtained on the whole matrix Unique spectral fingerprint Detects markers and spectral effects of adulteration Authenticity proof Allows to quantify key-quality parameters Allows to confirm botanical origins of mono-floral honey Alternative to conventional methods 20
21 Wine analysis by untargeted 1H-NMR Classification Country Appelation Vintage Variety 21
22 Another example: caviar authenticity Protection of the new PGI Caviar d Aquitaine 22
23 State of the art Even if these methods are not yet official, some food sectors use the 1H-NMR untargeted approach for their daily controls in our lab: Fruit Juice (supported by SGF) Honey (method presented at Apimondia / IHC workshop) Coffee (tested by several coffee associations) Milk (batch comparison, adulteration and degradation) Spices (adulteration saffron, curcumin ) Wine (discussion with OIV) Caviar (French trade association) 23
24 Conclusion: our best offer to fight against food adulteration: stable isotopes and NMR Stable isotope analysis constitutes a powerful tool for product authentication A number of routine applications exist (wines, spirits, fruit juices, honey, flavours, ) Recognized as official methods by EU / CEN / AOAC / OIV / AIJN. Used by public control laboratories But isotopic techniques also have limitations... The sample preparation is limited by the need for large amounts of pure compounds to be isolated without isotopic fractionation Need for large and up-to-date databases Untargeted analysis : from R&D to official control Consolidation of database Standardisation of the statistical approach Need for officialisation 24
25 Thanks for your attention We see only what we want to see or what we can see at the moment Freddy THOMAS 25
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