Chemical Contaminants & Residues in Food Community Subgroup ‘Unknowns’

1. Chemical Contaminants & Residues in Food CommunitySubgroup ‘Unknowns’ Collaborative Study for Determination of Melamine Report Sept. 2009 – Sept. 2010 124th AOAC Meeting & Exposition Orlando, FL 26-29 September, 2010

2. NH2 N N H2N NH2 N Chemical Contaminants & Residues in Food CommunityCollaborative Study for Determination of Melamine Renal failure in pets in the Republic of Korea (2004) Renal failure in pets in the United State of America (2007) Kidney stones in infants fed with Sanlu products (Sept. 2008) Deliberate addition to boost apparent protein content

3. Chemical Contaminants & Residues in Food CommunityCollaborative Study for Determination of Melamine WHO / Codex Alimentarius Maximum levels for melamine in food: ■ 1 mg/kg in powdered infant formula ■ 2.5 mg/kg in foods and animal feeds

4. Chemical Contaminants & Residues in Food CommunityCollaborative Study for Determination of Melamine 123rd AOAC Meeting & Exposition (Sept. 13-16, 2009, Philadelphia, PA) ■Session ‘Fraud in Food and Drug Safety’ ■Session ‘The Threat of the Unknown Contaminants: When You Don’t Know and What You Don’t Know’ ■Session ‘Finding Non-Target Chemical Contaminants in Food Stuff’ ■Hot topicrountable‘Dealingwith the Unknown – RiskAssessment and Risk Management Strategies of Industry and Regulators’ ►Initiate a project for a Collaborative Study to quantifymelamine in foodstuff

5. Chemical Contaminants & Residues in Food CommunityCollaborative Study for Determination of Melamine ■November 2009 People interest ■January 2010 Call for methods

6. Chemical Contaminants & Residues in Food CommunityCollaborative Study for Determination of Melamine Method Performance Criteria Purpose: Method for the quantitative determination of melamine (and cyanuric acid) in foodstuffs. The purpose of the method is to provide food industry and authorities with a method able to control melamine adulteration and monitor low levels in food. Applicability:Milk, dairy products, infant formula as well as food raw materials with a high risk of economically driven adulteration. Analytes: melamine (+cyanuric acid) Limit of detection: 0.05 mg/kg Regulatory limits: 1 mg/kg in milk-powder infant formula, and 2.5 mg/kg in other foods Accuracy (method recovery): 100% Selectivity (specificity): Should be able to differentiate melamine from cyanuric acid Repeatability (reproducibility): < 15% Technology: Mass spectrometry

7. Chemical Contaminants & Residues in Food CommunityCollaborative Study for Determination of Melamine Methods ■ HPLC-DAD by Ehlinget al. for melamine (MEL), cyanuricacid (CA), ammeline & ammelide ■ GC-MS by Litzauet al. for MEL, CA, ammeline & ammelide ■ LC-MS/MS by Smoker & Krynitsky for MEL & CA ■ LC-MS/MS by Desmarchelieret al. for MEL & CA

8. Chemical Contaminants & Residues in Food CommunityCollaborative Study for Determination of Melamine

9. Chemical Contaminants & Residues in Food CommunityCollaborative Study for Determination of Melamine

10. Chemical Contaminants & Residues in Food CommunityCollaborative Study for Determination of Melamine

11. Chemical Contaminants & Residues in Food CommunityCollaborative Study for Determination of Melamine

12. Chemical Contaminants & Residues in Food CommunityCollaborative Study for Determination of Melamine

13. Chemical Contaminants & Residues in Food CommunityCollaborative Study for Determination of Melamine ■Spikedsamples* Powdered infant formula Egg powder Soya protein Cereal and/or dry vegetables Driedpowderedshrimps ■Incurredsamples Driedmushrooms Feed * Eachsampleat four levels: blank, below maximum limit (ML), ML, above ML

14. Chemical Contaminants & Residues in Food CommunityCollaborative Study for Determination of Melamine Thierry DELATOUR Group Authenticity & Adulteration DepartmentQuality & Safety Nestlé Research Centre Vers-chez-les-Blanc 1000 Lausanne 26, Switzerland Phone: +41.21.785.9220 E-mail: thierry.delatour@rdls.nestle.com