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Rapid methods for feed safety: the CONffIDENCE project
Rapid Methods Conference & Open Day
25- 27 January 2010
www.conffidence.eu
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CONffIDENCE in a nutshell
CONtaminants in food and feed:
Inexpensive DEtectioN
for Control of Exposure
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CONffIDENCE, a European approach
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CONffIDENCE passport
FP7 Collaborative Project first call “Food, Agriculture & Fisheries, and Biotechnology”
Duration: May 2008 – April 2012 16 partners from 10 countries, representing
universities, research institutes, industry and SMEs
Budget: 7.5 Mio € Co-ordinator: RIKILT - Institute of Food
Safety, part of Wageningen UR (NL)
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Why CONffIDENCE ?
To assure chemical safety and quality in the European food supply; support of EC policies and competitiveness of food and feed industries
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Why CONffIDENCE ?
To improve multi-detection (“multiplex”) possibilities, e.g. for antibiotics
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Why CONffIDENCE ?
To improve inexpensive screening possibilities, e.g. for metal speciation
OH OH
OH
As As
O
OH
OHOH
Inorganic arsenic
Arsenous acidAs(III)
Arsenic acidAs(V)
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Why CONffIDENCE ?
To speed-up analysis for
factory approval of lots
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Why CONffIDENCE ?
To contribute to the assessment of risks of emerging contaminants e.g. plant toxins such as pyrrolizidine alkaloids
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Why CONffIDENCE ?
To contribute to the generation of data for exposure assessment, e.g. for PFC’s:
Perfluorooctane sulfonate (PFOS), perfluorooctanoic acid (PFOA) and their saltsScientific Opinion of the Panel on Contaminants in the Food chainAdopted on 21 February 2008
“Due to the substantial lack of suitable analytical data, many assumptions have been made in order to derive exposure estimates.”
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The objectives
Development and validation of new simplified inexpensive detection methods for chemical contaminants from farm to fork
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The objectives
Improved exposure assessment through monitoring of selected contaminants
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The objectives
Contribute to validation of predictive hazard behaviour models
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The objectives
Dissemination and training of new detection methods to all relevant stakeholders, to advance technology exploitation
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Food & Feed Fish/shellfish Fish feed Cereals Cereal-based feed
Potatoes/vegetables Honey Eggs Meat Dairy products
The commoditiesBased on their relevance for consumer safety and trade, the following target commodities were selected:
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The target contaminants
Persistent Organic Pollutants (POPs): - dioxin-like PCBs- brominated flame retardants- polycyclic aromatic hydrocarbons (PAH)
The targeted analytes represent a balance of emerging contaminants, high frequency detected residues as well as compounds which are not satisfactorily covered by current analytical methods.
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The target contaminants
Persistent Organic Pollutants (POPs) Perfluorinated compounds (PFCs):
- PFOS- PFOA
The targeted analytes represent a balance of emerging contaminants, high frequency detected residues as well as compounds which are not satisfactorily covered by current analytical methods.
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The target contaminants
Persistent Organic Pollutants (POPs) Perfluorinated compounds (PFCs) Pesticides:
- paraquat- diquat- dithiocarbamates
The targeted analytes represent a balance of emerging contaminants, high frequency detected residues as well as compounds which are not satisfactorily covered by current analytical methods.
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The target contaminants
Persistent Organic Pollutants (POPs) Perfluorinated compounds (PFCs) Pesticides Veterinary drugs:
- antibiotics, e.g. tetracyclines- coccidiostats, e.g. ionophores
The targeted analytes represent a balance of emerging contaminants, high frequency detected residues as well as compounds which are not satisfactorily covered by current analytical methods.
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The target contaminants
Persistent Organic Pollutants (POPs) Perfluorinated compounds (PFCs) Pesticides Veterinary drugs Heavy metals speciation:
- inorganic arsenic- methyl mercury
The targeted analytes represent a balance of emerging contaminants, high frequency detected residues as well as compounds which are not satisfactorily covered by current analytical methods.
OH OH
OH
As As
O
OH
OHOH
Inorganic arsenic
Arsenous acidAs(III)
Arsenic acidAs(V)
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The target contaminants
Persistent Organic Pollutants (POPs) Perfluorinated compounds (PFCs) Pesticides Veterinary drugs Heavy metals speciation Biotoxins:
- plant alkaloids: pyrrolizidine, tropane, ergot - marine biotoxins (PSP, DSP) - mycotoxins: DON, ZEA, T-2/HT-2, FB1, FB2
The targeted analytes represent a balance of emerging contaminants, high frequency detected residues as well as compounds which are not satisfactorily covered by current analytical methods.
Ergot bodies
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The technologies
Bio-analytical techniques
MS-based techniques
Spectroscopic techniques
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Bio-analytical techniques
Analyte Bio-molecule
Platform
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Bio-analytical techniques
Antibodies
Receptors
Transport proteins
Aptamers
Dipsticks
SPR
Flow cytometry
Electrochemical
immunosensors
Cytosensors
Bio-molecule Platform
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Connection with the potentiostat equipment
ne-
HQ
HRP
red ox
ox
H2O H2O2
magnet
SFM-HRPSFM
A
mGEC sensor
red
i
ii iii
iv
i
Connection with the potentiostat equipment
ne-
HQ
HRP
red ox
ox
H2O H2O2
magnet
SFM-HRPSFM
A
mGEC sensor
red
i
ii iii
iv
i
The technologies
Multiplex dipsticks assays
Multiplex SPR biosensor
Multiplex flow cytometry
Electrochemical immunosensors
Cytosensors
D) Chloramphenicol
C) Tylosin
B) Quinolones
A) Sulfonamides
Transcription
Translation
mRNA
Reporterplasmid
PLuciferaseoperon
Luciferase
Luminescence
R
P = promoterR = regulatory
protein
Bio-analytical techniques
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MS-based techniques
Simplified mass spectrometric (MS) methods
LC-MS or LC-MS/MS
DESI-MS
MS-based techniques
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Near Infrared (NIR) imaging methods
(nombre de pixels en x)(n
ombr
e de
pix
els
en y
)(Wavelength nm)
Spectroscopic techniques
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Project structure
WP1a Persistant organic
pollutants (POPs) Fish, fish feed and cereal
based food Flow cytometry based
multiplex assay GCxGC-TOF-MS
comprehensive screening
ICT-RIKILT-JRC IRMM-NUTRECO
WP1b Perfluorinated
compounds (PFCs) Fish, fish feed and dairy
products Simplified LC-MS method
CSIC-ICT-JRC IRMM-NUTRECO
WP1c Pesticides (Paraquat,
Diquat) Potato and cereals Validated electrochemical
magneto immunosensor assay
DESI-MS / DART-MS (dithiocarbamates)
RIKILT-CSIC-FERA-CVUA-ICT
WP2a Coccidiostats Eggs, Laying hens feed Flow cytometry based
multiplex immunoassay
JRC IRMM-CER-RIKILT-QUB-NUTRECO-FERA
WP3 Heavy metals Marine based feed and
food Cytosensor and solid
phase extraction approaches
DTU Food-TUT-JRC IRMM-NUTRECO-RIKILT
WP2b Antibiotics Fish, feed, meat, honey Single component and
multiplex dipstick Electrochemical magneto
immunosensor assay
FERA-UNISENSOR-CER-CSIC-NESTLE-JRC IRMM-NUTRECO
WP4a Alkaloids (pyrrolizidine,
tropane and ergot) Honey, feed and cereals Validated multiplex
dipstick assay On-line imaging method
RIKILT-QUB-CRAW-UNISENSOR-FERA-CER-NUTRECO
WP4b Marine biotoxins Shellfish Rapid multiplex SPR
biosensor assay
QUB-USC-CER-JRC IRMM
WP4c Mycotoxins Cereals for food and feed Validated multiplex
dipstick
ISPA-UNISENSOR-CER-RIKILT-NUTRECO-NESTLE
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Project structure
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Dissemination: the website
www.conffidence.eu
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Website: some statistics (Google Analytics)
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33 events on-line 15 news on-line3 newsletters delivered14 links to related projects Publications1 Open DayTo be informed, registration on-line48 stakeholders registered
Information for stakeholders
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First results
Multiplex immunoassay for coccidiostats
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First results flow cytometry
Calibration curves of
nicarbazin, diclazuril,
salinomycin and narasin
in buffer
Sensitivities of nicarbazin, diclazuril and salinomycin assays promising
Multiplex immunoassay for coccidiostats
nicarbazin
salinomycin
narasin
diclazuril
20
40
60
80
100
0.001 0.01 0.1 1 10 100 1000
Nicarbazin
Salinomycin
Narasin
Diclazuril
0.0001
ng/ml
B/B0
20
40
60
80
100
0.001 0.01 0.1 1 10 100 1000
Nicarbazin
Salinomycin
Narasin
Diclazuril
0.0001
ng/ml
B/B0
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First results
Multiplex immunoassay for coccidiostats Tetracycline Tetrasensor assay for feed
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Tetracycline Tetrasensor assay for feed
- Pig FEED - dilution (10x) in buffer- Homogenisation (2 min)- Centrifugation (2 min)- 200 µl of supernatant + reagents- Test : 10 min at RT
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First results
Multiplex immunoassay for coccidiostats Tetracycline Tetrasensor assay for feed Ergot detection in cereals by NIR imaging
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Ergot detection in cereals by NIR imaging
Pixels
Pix
els
Escourgeon plaque 3_01logxCbpfmsg0sg1.spf
50 100 150 200 250 300
50
100
150
200
Barley sample including an ergot body
Image rebuilt at 1200 nm after 1st Derivative preprocessing
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Ergot detection in cereals by NIR imaging
Discrimination Ergot/Wheat based on 2 wavelengths (1220 nm/ 1440 nm)
-0.24 -0.22 -0.2 -0.18 -0.16 -0.14 -0.12 -0.1 -0.08 -0.06 -0.040.02
0.04
0.06
0.08
0.1
0.12
0.14
0.16
0.18
0.2
1220 nm
1440
nm
Ergot (Cal)Wheat (Cal)
95% confidence ellipse
95% confidence ellipse
Ergot (Val)Wheat (Val)
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Acknowledgements
The CONffIDENCE project is financially supported by the European Commission under Grant Agreement no. 211326
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Acknowledgements
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More information
Website: www.conffidence.euContact:
e-newsletter
(registration on website)