rapid methods for feed safety: the conffidence project rapid methods conference & open day 25-...

Rapid methods for feed safety: the CONffIDENCE project 

Rapid Methods Conference & Open Day

25- 27 January 2010

www.conffidence.eu

CONffIDENCE in a nutshell

CONtaminants in food and feed:

Inexpensive DEtectioN

for Control of Exposure

CONffIDENCE, a European approach

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)

Why CONffIDENCE ?

To assure chemical safety and quality in the European food supply; support of EC policies and competitiveness of food and feed industries

Why CONffIDENCE ?

To improve multi-detection (“multiplex”) possibilities, e.g. for antibiotics

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)

Why CONffIDENCE ?

To speed-up analysis for

factory approval of lots

Why CONffIDENCE ?

To contribute to the assessment of risks of emerging contaminants e.g. plant toxins such as pyrrolizidine alkaloids

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.”

The objectives

Development and validation of new simplified inexpensive detection methods for chemical contaminants from farm to fork

The objectives

Improved exposure assessment through monitoring of selected contaminants

The objectives

Contribute to validation of predictive hazard behaviour models

The objectives

Dissemination and training of new detection methods to all relevant stakeholders, to advance technology exploitation

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:

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.

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.

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.

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.

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)

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

The technologies

Bio-analytical techniques

MS-based techniques

Spectroscopic techniques

Bio-analytical techniques

Analyte Bio-molecule

Platform

Bio-analytical techniques

Antibodies

Receptors

Transport proteins

Aptamers

Dipsticks

SPR

Flow cytometry

Electrochemical

immunosensors

Cytosensors

Bio-molecule Platform

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

MS-based techniques

Simplified mass spectrometric (MS) methods

LC-MS or LC-MS/MS

DESI-MS

MS-based techniques

Near Infrared (NIR) imaging methods

(nombre de pixels en x)(n

ombr

e de

pix

els

en y

)(Wavelength nm)

Spectroscopic techniques

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

Project structure

Dissemination: the website

www.conffidence.eu

Website: some statistics (Google Analytics)

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

First results

Multiplex immunoassay for coccidiostats

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

First results

Multiplex immunoassay for coccidiostats Tetracycline Tetrasensor assay for feed

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

First results

Multiplex immunoassay for coccidiostats Tetracycline Tetrasensor assay for feed Ergot detection in cereals by NIR imaging

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

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)

Acknowledgements

The CONffIDENCE project is financially supported by the European Commission under Grant Agreement no. 211326

Acknowledgements

More information

Website: www.conffidence.euContact:

coordination@conffidence.eu

e-newsletter

(registration on website)