Food Authenticity Profiling Foods

Michal Godula

What is Food Authenticity?

Food authenticity is about misrepresentation by either mislabeling or by adulterating usually with lower cost material Typical examples: Wine authenticity illegal sugar addition to grape juice Addition of sugar to honey Addition of hazelnut oil to olive oils Mislabeling geographical origin (wine, olive oil) Mislabeling organic food products Adulteration of meat with cheaper species
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Major Food Authentication Schemes and Systems

Protected Designation of Origin (PDO) covers agricultural products and foodstuffs which are produced, processed and prepared in a given geographical area using recognized know-how.

Council Regulation (EC) No 510/2006 of 20 March 2006

Protected Geographical Indication (PGI) covers agricultural products and foodstuffs closely linked to the geographical area. At least one of the stages of production, processing or preparation takes place in the area. Traditional Speciality Guaranteed (TSG) highlights traditional character
http://ec.europa.eu/agriculture/quality/

Council Regulation (EC) No 509/2006 of 20 March 2006

DOOR Database

More than 1200 entries Different countries participate Number of different foods listed Typically: olive oils, wines, cheeses, honey, beer etc

Food Authentication Challenges

Chemically identical foods or identical chemical entities Unique marker compounds rarely found - more often small analytical differences (isotopic patterns) Large natural variability based on climatic conditions, fertilizers used, variety, processing Techniques must be able to distinguish small differences Databases of authentic foods must be available to understand natural variability
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Which Techniques Can be Applied?

Technique Infrared spectroscopy (NIR, MIR) Raman spectroscopy Nuclear Magnetic Resonance Atomic absorption and emission spectroscopy, ICP/MS Mass spectrometry (GC/MS, LC/MS) Chromatography (HPLC/RID) IRMS Parameters measured Spectral profile, characteristic frequencies Spectral profile, characteristic frequencies Spectral profile, characteristic frequencies Elemental composition Volatile components, phenolic compounds Amino acids, carbohydrates, phenolic compounds Stable Isotope Ratios

Other Approaches
DNA based methods like PCR used mainly for adulteration of meat and meat based products - addition of chicken and pork meat into minced beef - differentiation of cattle breed - adulteration of foie gras with chicken* Immunology based approach most of the techniques based on ELISA detection of specific proteins - presence of vegetable proteins in milk or meat - adulteration of sheep/goat milk with cow milk

7 * L.M. Reid et al. / Trends in Food Science & Technology 17 (2006) 344353

Data Analysis: Chemometric Approach

Typically large number of variables involved, ie delta value, elemental composition, mass spectra, compound concentration etc Multivariate analysis must be applied Supervised or unsupervised methods Unsupervised: classification of sample without any knowledge about the origin Supervised: similarity of unknown to authentic material Choosing appropriate approach is the key to successful analysis

Gonzalves, A, Trends in Analytical Chemistry, Vol. 28, No. 11, 2009

Chemometric Methods Used

Unsupervised methods ANOVA MANOVA Principal Component Analysis (PCA) - reduces dimensionality Cluster Analysis (CA) - grouping of samples based on their similarity Supervised methods (discriminant techniques) Linear discriminant analysis (LDA) - characterize or separate two or more classes Partial Least Squares (PLS) - fundamental relations between two matrices Artificial Neural Networks (ANN) - complex relationships modelling

Isotopes - Sources of Variation

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Climatic Factors Affecting D/H Ratio

Evaporation & Reprecipitation

H2O HDO H2O HDO

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Interpolated 2H of Precipitation

Produced by James Elhringer (University of Utah)

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IRMS Information Delivered

Precise Isotope Ratios Element Hydrogen Carbon Nitrogen Oxygen Sulfur others minor Isotope
2H

natural abundance [%] 0.01557 1.11140 0.36630 0.20004 4.21500

(D) 13C 15N 18O 34S

Thats where the information is

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Authenticity Control of Food and Beverages

CSIA H2 N2 CO CO2
D
15N 18O 13C 18O

Compound Specific Isotope Analysis

Multiple Element BSIA Isotope Ratio Mass Spectrometry

Bulk Stable Isotope Analysis

GasBench
Gas Specific Isotope Analysis

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Different Approaches to IRMS

BSIA
Bulk Stable Isotope Analysis
Ref. gases

Reactor
Open Split

DELTAplusXP GC Separation 2H and 18O

or

Elemental Analyzer Conversion

13C and 15N and 34S

CSIA
Compound Specific Isotope Analysis

Ref. gases

Capillary Reactor GC Separation of complex mixtures

On-line Conversion
Open Split 2H

DELTAplusXP or 18O or 13C or 15N of all Compounds

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Sample Introduction Conversion to Simple Gases

BSIA: Elemental Analyzer (EA) for C, N, S BSIA: High temperature conversion (TC/EA) for C, H CSIA: High temperature conversion (GC/TC) for C,H CO2, SO2, N2 CO2, H2 CO2, H2

18O
y TC/EA >1400 C n-y
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CO
x/2

2H

Cn Hx Oy

H2

irm GC/MS

HD

0.015 % 99.985 %

H2

Principles of Multicollector IRMS

Multi Collectors for Simultaneous Detection H2 2 N2 2 CO O2 2 CO2 2 SO2 2
under continuous flow conditions 4He is about 107 times more abundant than HD magnet m/z 4

m/z 3 (HD) m/z 2 (H2)

with energy filter

ion source

universal triple collector for N2, CO, O2,CO2, SO2

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Compound Specific Isotope Analysis - GC Isolink

Advantages of irm-GC/MS with GC-ISoLink Allows simultaneous determination of C, N and H within one sequence Combustion (C, N) x high temperature conversion (H) Provides chromatographic profile of the sample AND isotopic ratio information

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CSIA Examples Tequila Fingerprinting by GC/C-IRMS

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Honey Adulteration
Problem: addition of cheap sugars to honey Current analytical approach: EA-IRMS to assess 13C value of honey and its protein fraction (limit cca 7% of C4 sugar addition)

Solution: irm-LC/MS using IsoLink interface allows: - comparison of 13C value of different sugars - determination of the sugar pattern

TMO application note 30024 or Testing honey adulteration by 13 C-EA/LC-IRMS, Elflein L., Apidologie (2008),

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Honey Adulteration Examples

Honey 5

Honey 1
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Honey 2

TMO appnot e30024

Assessing the Origin of Cheeses in Europe

Measurands taken: Stable isotope ratios, major, trace and radioactive elements Techniques used: IRMS, AAS, ICP-MS Cheese

Lyophilization Extraction H2O

Fat extraction Hydrolysis to glycerol

Lyophilization Extraction, H2O pH 4.3 Dry insoluble fraction

Wet Digestion Dilution

IRMS
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ICP/MS, AAS

Pillonel L et al, Lebensm.-Wiss. u.-Technol. 36 (2003) 615623

Assessing the Cheese Origin in Europe

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Pillonel L et al, Lebensm.-Wiss. u.-Technol. 36 (2003) 615623

Infrared Spectroscopy
Benefits: Ease of use Fast response No or limited sample preparation Low price Limitations: Lower sensitivity Small spectral differences Typical examples: Near Infrared Spectroscopy (NIR, FT-NIR) Different Sampling accessories: Sample cup spinner heterogeneous materials, SabIR Fiber Optic Probe material identification, Viscous Liquid Sampler syrups, suspensions, Softgel detector (Gum), Fiber Optic probes in-process testing of solids and liquids
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FT-NIR to Assess Geographic Origin of Olive Oils

Antaris FT-NIR with sample cup spinner Direct measurement in 2mm path length cell 4 cm-1 resolution Spectral range - 10000 and 4500 cm-1

O. Galtier et al.; Analytica Chimica Acta 595 (2007) 136144

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FT-NIR to Assess Geographic Origin of Olive Oils

Quantification of fatty acids and triglycerols Classification in registered designation origin

O. Galtier et al.; Analytica Chimica Acta 595 (2007) 136144

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Pros/Cons of Different Analytical Approaches

Methods

Sample preparation costs

Speed

Investment and operating costs

Probability of success

NMR

IRMS

Chromatography GC/MS, LC/MS

Infrared Spectroscopy (NIR. IR, Raman)

Elemental analysis (ICP/MS, AAS, ICP)

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Which Technique is the Most Appropriate?

Isotope Ratio Mass Spectrometry considered as standard approach Organic mass spectrometry gaining popularity Elemental analysis can provide additional information to IRMS Non-destructive techniques offer high speed and almost no sample preparation (FTNIR, FTIR, Raman) Sophisticated data analysis - chemometric approaches Thermo Scientific offers solution for all authenticity analysis approaches

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