federica camin
TRANSCRIPT
Stable isotope ratio analysis for protecting foods with
denomination
Federica Camin
Strategies
paper traceability
(EU Reg. 178/2002)
analytical tests
robust, validated, official methods
(EU Reg, OIV, ISO/ENV, AOAC)
robust, validated, official limits or reference data
Stable Isotope Ratios of Bioelements
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2H/1H
13C/12C
15N/14N
18O/16O
34S/32S
Stable isotope ratios methods as official standards
Year Method product Method Isotope Ratio Fraud
1987 OIV wine, must SNIF-NMR D/H sugar addition (beet, cane)
1990 EU Reg 2676/90, encl. 8 wine, must SNIF-NMR D/H sugar addition (beet, cane)
1991 AOAC 998,12 honey IRMS 13C/12C sugar addition (cane)
1993 ENV 12140, 13070 fruit juice IRMS 13C/12C sugar addition (cane)
1995 AOAC 995,17 fruit juice SNIF-NMR D/H sugar addition (beet, cane)
1996 OIV 2/96 wine, must IRMS 18O/16O addition of water/mislabelling
1997 EU Reg 2676/90, 822/97 wine, must IRMS 18O/16O addition of water/mislabelling
1997 ENV 12141 fruit juice IRMS 18O/16O addition of water/mislabelling
2000 AOAC 2000.19 mapple syrup SNIF-NMR D/H sugar addition (beet, cane)
2000 OIV 71/2000 vinegar SNIF-NMR, IRMS D/H, 13C/12C sugar addition (beet, cane)
2001 OIV 17/2001 wine, must IRMS 13C/12C sugar addition (cane)
2003 EU Reg. 2676/90, 440/03 wine, must IRMS 13C/12C sugar addition (cane)
2003 OIV MA-F-AS314-03 wine IRMS 13C/12C technogenic CO2
2004 AOAC 2004,01 fruit juice, maple syrup SNIF-NMR D/H sugar addition (beet, cane)
2006 AOAC 2006,05 vanillin SNIF-NMR D/H synthetic vanillin
2007 OIV-MA-AS312-07 wine IRMS 13C/12C addition of glycerol
2011 EU Reg 584/2011 Grana Padano DOP IRMS D/H, 13C/12C, 15N/14N, 34S/32S mislabelling
2013 EN 16466-1, 2, 3 vinegar SNIF-NMR, IRMS D/H, 13C/12C, 18O/16O water and sugar addition (beet, cane)
2013 OIV 510, 511/2013 vinegar IRMS 13C/12C, 18O/16O water and sugar addition (cane)
520 denomination for Italian wines (DOC/DOCG/PDO/IGP) Wine
OIV-MA-AS311-05 for site-specific analysis of the D/H ratio using 2H-SNIF-NMR (2H-Site-specif Natural Isotope Fractionation - Nuclear Magnetic Resonance)
OIV-MA-AS312-06 for analysis of the 13C/12C ratio (expressed as d13C‰) using Isotope Ratio Mass Spectrometry (IRMS)
OIV-MA-AS2-12 for analysis of the 18O/16O ratio (expressed as d18O‰) using Isotope Ratio Mass Spectrometry (IRMS).
Databank: EU reg 555/2008
Wine Databank EC Reg. Nº 2729/2000
Malta
4
Slovakia
15
Slovenia
20
Hungary
50 Czech Rep.
20
Cyprus
10
France
400
Germany
200
Italy
400
Spain
200 U. Kingdom
4
Austria
50
Greece
50
Luxembourg
4
Portugal
50
Wine data Maintenance of databank
Arbitration of disputes
Analysis of samples
Development and
validation of methods
Validation of data
Training
EC Reg. Nº 2120/2004
Romania
70
Bulgaria
30
EC Reg. Nº 2030/2006
Definition of limits and compliance
selection of reference data
computing 95% reference limits (t-student two tailed distribution)
c) the result is outside the limit, but, adding the measurement uncertainty, is inside the limit
d) the result and uncertainty are outside the limit
Situations a and b do not cause any consequences since the results are in compliance with the
a) b) c) d)
Upper limit
Lower limit
= result
= uncertainty
Balsamic vinegar of Modena PGI (Aceto balsamico di Modena)
obtained by mixing wine vinegar, cooked or concentrated grape must and caramel.
PGI status (Protected Geographical Indication) with EC Regulations 583/2009
6° PDO/PGI Italian product for sales (260 million € /year)
The geographical origin of vinegar and must is not specified
Modena
Stable Isotope ratio analysis in vinegar
CEN (European Commitee for Standardisation) from 01/03/2013 EN 16466-1 D/H SNIF NMR acetic acid EN 16466-2: 13C/12C IRMS acetic acid EN 16466-3: 18O/16O IRMS water
OIV from 06/2013
RISOLUZIONE OIV-OENO 510-2013: 13C/12C IRMS acetic acid
RISOLUZIONE OIV-OENO 511-2013: 18O/16O IRMS water
Dordevic et al., 2013
(D/H)1 (D/H)2 δ13C δ18O
Wines, 95% CI 98.8 / 106 124.5 / 135.5 -29.3 / -24.3 -1.3 / 8.9
Beet sugar 92.5 -27.5
Cane sugar 109.5 -12
Water (north Italy) -9
Water (south Italy) -5
Stable Isotope ratios for the autenticity of ABM
-34
-32
-30
-28
-26
-24
-22
-20
-18
-16
-14
-12
-10
90 92 94 96 98 100 102 104 106 108 110 112 114 116 118 120
(D/H)I ppm ethanol
d13C
‰ e
than
ol
beet sugar
synthesis
cane sugar
Wine/must/
vinegar
(D/H)I
(D/H)II
The “top ten” Italian PDO and PGI for sales 2012
milions €
Grana Padano 1.395
Parmigiano Reggiano 1.357
Prosciutto di Parma 992
Prosciutto di San Daniele 302
Mozzarella di Bufala Campana 288
Aceto Balsamico di Modena 260
Gorgonzola 249
Mortadella Bologna 224
Bresaola della Valtellina 215
Mela Alto Adige 170
Total (production costs) 5.453
% of total 83,8
Source: Osservatorio Ismea prodotti Dop e Igp
Random Forest Model (2011) for the traceability of Grana Padano cheese
98.5% correct classification, in cross validation
Validation of the methods
International collaborative study:
IUPAC protocol and the ISO Standards 5725/2004 and 13528/2005.
7 types of cheeses in blind duplicate
20 laboratories
13 labs answered
Sr and SR are available
Camin et al., RCM, 2015
Specification for PDO Grana Padano cheese
Isotopic ratios as official parameters for verifying the authenticity of PDO Grana Padano cheese in grated and shredded form
The variability ranges have been deposited at the Italian Ministry of Agriculture, Food and Forestry
Passate di pomodoro made in Italy
D.M. 23 settembre 2005 (GU n. 232 del 5 ottobre 2005):
‘Definizione di passata di pomodoro: la denominazione di vendita ‘Passata di Pomodoro’ è riservata al prodotto ottenuto direttamente dal pomodoro fresco, sano e maturo…, per spremitura, eventuale separazione di bucce e semi e parziale eliminazione dell’acqua di costituzione in modo che il risiduo ottico rifrattometrico risulti compreso tra 5 e 12 gradi Brix ….
Non è consentito concentrare il succo di pomodoro al di sopra dei 12 gradi brix e provvedere alla successiva diluizione per la preparazione della passata di pomodoro…
Per la determinazione dell’acqua aggiunta ai prodotti che riportano tale denominazione di vendita … si applica il metodo di analisi UNI ENV 12141 (1997) e successive modifiche = Determinazione del rapporto 18O/16O nei succhi di frutta e vegetali
Bontempo et al., Food Control 2014
d18O passate > 7,5°Bx da evaporazione
Bontempo et al., Food Control 2014
Limite: -1.4‰
Conclusion
SIRA, sometime in combination with other analytical techniques, is able to detect the authenticity of food with denomination:
DOC/DOCG/PDO wine
Aceto Balsamico di Modena PGI
Grana Padano and Parmigiano Reggiano PDO
passata ‘Made in Italy’
Other examples: Extravirgin olive oil PDO, PGI, PDO ham, organic/conventional products, …
Thank you for your kind attention!
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