Analysis of isotopic ratios for the detection of illegal...
Transcript of Analysis of isotopic ratios for the detection of illegal...
Giovanni CalderoneClaude Guillou
Giovanni CalderoneGiovanni CalderoneClaude GuillouClaude Guillou
Analysis of isotopic ratios for the detection of illegal watering of
beverages
Analysis of isotopic ratios for the Analysis of isotopic ratios for the detection of illegal watering of detection of illegal watering of
beveragesbeverages
Commission of the European Communities, Joint Research CentreInstitute for Health and Consumer Protection
Physical & Chemical Exposure Unit - BEVABS 21020 Ispra (VA) Italy
Physical & Chemical Exposure Unit
27-28 Mars 2006 – 4th International Workshop on Water in Food, Brussels BEVABS
Stable isotopes(non radioactive isotopes)
Organic matter is basically composed of H, O, C and N.
1213
1816
17
All of these elements are distributed in isotopes (= atomic number, ≠ mass), whose amount is influenced by natural processes and human activities.
27-28 Mars 2006 – 4th International Workshop on Water in Food, Brussels BEVABS
Stable isotopes
Element Isotope Relative abundance (%)
1H 99.9855
2H (D) 0.0145
12C 98.892
13C 1.108
14N 99.6337
15N 0.3663
16O 99.7586
17O 0.0375
18O 0.2039
Oxygen
Nitrogen
Carbon
Hydrogen
The measurements of the ratios (Xheavy / Xcommon) give information about the botanical (13C/12C), animal (15N/14N) or geographic origins (2H/1H, 18O/16O) of the products.
Many studies and applications:
Biomedical studies
•Environmental research
•Food authentication
•Fight against frauds
27-28 Mars 2006 – 4th International Workshop on Water in Food, Brussels BEVABS
Application of Isotope Ratios analysis in food authentication
Product Adulteration AnalysisFruit juices Watering (D/H) and 18O-IRMS
Fruit juices Sweetening 13C-IRMS and 2H-NMR
Aromas, flavours (vanillin, raspberry ketone, …)
Mislabelling
(artificial ↔ natural)13C-IRMS, 18O-IRMS, 2H-NMR
Alcoholic beveragesMislabelling(geographical and botanical origins)
13C-IRMS, 18O-IRMS, 2H-NMR, 2H- and 18O-pyr-IRMS
Wines Watering (D/H) and 18O-IRMS
Wines Chaptalisation (addition of sugars) 13C-IRMS, 2H-NMR
Honey Addition of inverted and cane sugars 13C-IRMS
O
OH
OCH3
HIRMS = Isotopic Ratio Mass Spectrometry2H-NMR = Nuclear Magnetic Resonance of Deuterium
27-28 Mars 2006 – 4th International Workshop on Water in Food, Brussels BEVABS
Various factors influencing the oxygen isotope composition of organic matter.Source Epstein et al.,1977
Deuterium and oxygen
27-28 Mars 2006 – 4th International Workshop on Water in Food, Brussels BEVABS
Deuterium and oxygen: geographic origin
• Main source H and O in organic matter is the water• 2H and 18O fractionation phenomena are associated to Temperature
→ Climate and geography affect 2H and 18O distribution
CONTINENTδ 18O ≡ 0 ‰δ 2H ≡ 0 ‰
27-28 Mars 2006 – 4th International Workshop on Water in Food, Brussels BEVABS
Geographical distribution of 18O(meteoric waters 1992/93)
Source: K. Rozanski, L. Araguas, R. Gonfiantini – Isotopic patterns in modern global precipitation. In: Continental Isotope Indicators of Climate, American Geophysical Union Monograph, 1993
27-28 Mars 2006 – 4th International Workshop on Water in Food, Brussels BEVABS
How to measure 18O/16O ratio of water
T cost.16 16 1618 182 2 2
(8-12 h at 25 °C) (m=46)(m=44)C O + H O C O O + H O
⎯⎯⎯⎯⎯→←⎯⎯⎯⎯⎯
Water is equilibrated with CO2 gas and compared to a a reference H2O
Headspace
(CO2+H2O
+EtOH)
Cryogenic Trap
(-H2O, -EtOH)IRMS
S. Epstein, T. K. Mayeda – Variation of 18O content of waters from natural sources. Geochimica et CosmochimicaActa, 1953, 213-224.
27-28 Mars 2006 – 4th International Workshop on Water in Food, Brussels BEVABS
Magnet
m/z 44 (12C16O2)
m/z 46(12C18O16O,
13C17O16O, 12C17O2)m/z 45(13C16O2, 12C17O16O)
Gas (CO2 )
How works IRMS system?
δ ‰VS SMOW
Ion source Mass detectors(Faraday cage)
27-28 Mars 2006 – 4th International Workshop on Water in Food, Brussels BEVABS
Example: use of isotope ratios for the detection of adulteration of wine
0,0
2,0
4,0
6,0
8,0
10,0
12,0
14,0
-30,0 -29,0 -28,0 -27,0 -26,0 -25,0 -24,0 -23,0 -22,0 -21,0 -20,0
δ 13C
δ 18
O
DB 96
DB 95
DB 97
Suspect samples A
Suspect samplesB
Suspicious wine Samples
AUTHENTIC WINES FROM FROM THE SAME COUNTRY THAN THE SUSPICIOUS SAMPLESDATA FROM EU WINE ISOTOPIC DB
27-28 Mars 2006 – 4th International Workshop on Water in Food, Brussels BEVABS
-10
-8
-6
-4
-2
0
2
4
6
-80 -70 -60 -50 -40 -30 -20 -10 0 10 20δ D ‰
δ 18O ‰
Fresh orange juices (Israel, Brasil,
Marocco)
Orange juices made from concentrate
Fresh grapefruit juice
Pineapple juice made from concentrate
Ispra Tap Water
Example: fruit juices
27-28 Mars 2006 – 4th International Workshop on Water in Food, Brussels BEVABS
International Official Methods
UE (European Union)
European Commission Regulation N° 2676/90 determining Community methods for the analysis of wines - Official Journal N° L 272/64. (SNIF-2H-NMR of ethanol from wine)European Commission Regulation N° 822/97 amending E.C. Regulation N° 2676/90 – Official Journal of the European Communities N° L117/10. (18O/16O of water in wine)European Commission Regulation N° 440/2003 amending E.C. Regulation N°2676/90 – Annex II, Official Journal of the European Union N° L66/15. (13C/12C of ethanol from wine)
OIV(Organisation International de la Vigne et du Vin,
International Organisation of Vine and Wine)
Resolution OENO 2/96 – Mouillage – Détermination du rapport isotopique 18O/16O de l'eau des vins.Resolution OENO 71/2000 – in Recueil des methodes d'analyse des vinaigres de vin, Annexe XX “Vinaigres De Vin -authentification par FINS-RMN ® et d'autres méthodes isotopiques”.Resolution OENO 17/2001 in Compendium on International Methods of Wine and Musts - “Determination by isotope ratio mass spectrometry of 13C/12C of wine ethanol or that obtained through the fermentation of musts, concentrated musts or grape sugar”.Resolution OENO 7/2005, completing the Annex C of the Compendium of International Methods of Analysis of Musts and Wines by the method for the “Determination of the carbon isotope ratio 13C/12C of CO2 in sparkling wines method using isotope ratio mass spectrometry (IRMS)”.
27-28 Mars 2006 – 4th International Workshop on Water in Food, Brussels BEVABS
International Official Methods (2)
CEN(Comité Européen de Normalisation)
CEN/TC174 N° 108, 1996: Determination of the stable carbon isotope ratio (13C/12C) of sugars from fruit juices - Method using Isotope Ratio Mass Spectrometry.CEN/TC174 N° 109, 1996: Determination of the stable oxygen isotope ratio (18O/16O) of water from fruit juices - Method using Isotope Ratio Mass Spectrometry.CEN/TC174 N° 110, 1996: Determination of the stable hydrogen isotope ratio (2H/1H) of water from fruit juices - Method using Isotope Ratio Mass Spectrometry.
AOAC(Association Of Analytical Communities)
AOAC method, detection of addition of C-4 Plant Sugars in Honey by 13C/12C analysis. Methods 978.17, 991.41 and 998.12.AOAC method, detection of addition of beet sugar in Fruit Juices. Method 995.17. (SNIF-2H-NMR)AOAC method, detection of addition of beet sugar in Fruit Juices. Methods 981.09 and 982.21. (13C/12C analysis)AOAC method, Carbon ratio mass spectrometric method for detection of corn syrup and cane sugar in maple syrup. Method 984.23.AOAC method, determination of sugar beet derived syrups in frozen concentrated orange juice-δ18O measurements in water. Method 992.09.AOAC method, Carbon stable isotope ratio of ethanol derived from fruit juices and maple syrups. Method 2004.01.
JRC - BEVABS
Bureau Européen de Vins, Alcools et Boissons Spiritueuses
European Office for Wines, Alcohols and Spirit Drinks
EU wine database and isotopic databank
Cyprus MaltaCzech Republic Slovakia
Hungary Slovenia
+
22HH--NMR NMR (1990)(1990) and and 1313CC--IRMS IRMS (2003)(2003) (ethanol)(ethanol)1818OO--IRMS IRMS (1997) (1997) (water)(water)
27-28 Mars 2006 – 4th International Workshop on Water in Food, Brussels BEVABS
Carbonated waters: 13C/12C of CO2
Two bottled water samples labelled as naturally sparkling:
«Acqua di Nepi», from Latium (Italy)measured δ13C = -1.1 ‰, literature value -1.3 ‰
Other bottled water, from Rome (Italy)measured δ13C = -42.1 ‰ !
27-28 Mars 2006 – 4th International Workshop on Water in Food, Brussels BEVABS
Thanks for your
attention!
Thanks for your
attention!