Real World-Application Methods From a Food Testing Laboratory

Transcription

1 Real World-Application Methods From a Food Testing Laboratory Sonja Riediker Nestlé Quality Assurance Center Singapore 21 March 2006

2 Overview Nestlé Quality Nestlé Laboratory Network LC-MS/MS in Routine Analysis LC-MS/MS: Advantages and Challenges Conclusions 44

3 Nestlé Products... A Diversity as the Consumers More than 130 different brands: 45 Beverages e.g. Nescafé, Milo, Nestlé Aquarel Milk products e.g. Nespray, Nido, Molico, LC 1 Nutrition Products e.g. Lactogen, Cerelac, PowerBar Culinary products e.g. Maggi, Buitoni, Thomy Chocolate, Confectionary e.g. Crunch, Cailler PetCare e.g. Purina Friskies, Felix Alcon Pharmaceutical company etc.

4 Nestlé Quality: General Approach People are involved all along the Supply Chain Agricultural Production Consumer Primary & Secondary Transformation Transport Production Transport Distribution Trade Upstream Downstream 46

5 Nestlé Quality: Three Dimensions Health Nutritional Value, Functionality Food Safety Regulatory Compliance Social Price/Performance Convenience Cultural, Conviviality, Pleasure Sensory Aspect, Colour Aroma, Flavour Texture, Sound 47

6 Nestlé Quality: Health Our approach: Usage of high-quality and safe ingredients Release parameters for production (physical, microbiological, and chemical) Monitoring parameters for due diligence On- and in-line quality control during production Quality control of finished products Release parameters (physical, microbiological, and chemical) Monitoring parameters for due diligence 48

7 Release and Monitoring Parameters Examples: Hygiene indicators and pathogens Contaminants e.g. pesticides, mycotoxins, veterinary drugs, processing contaminants, toxic metals, Authenticity (adulteration), GMO Compositional parameters e.g. vitamins, minerals, carbohydrates, fat content Claims e.g. low sodium, sugar-free, MSG-free 49

8 Nestlé Laboratory Network Factory Laboratories rapid tests or alternative methods, high throughput (on-, inline, on-site) Regional Laboratories accredited by local authorities, internal proficiency testing (Ptest) highly specialized or expensive analyses (e.g. HPLC-UVD, LC-MS/MS, ICP-MS, GC-ECD, etc) External Laboratories accredited, participation in international or internal proficiency testing 50

9 Nestlé Research Center (NRC) Quality and Safety Department Method development, modifications (ca. 250 Laboratory Instructions) Groups with focus on Contaminants Ingredients and Authenticity Packaging, Off-Flavour Microbiological Safety Chemical Food Safety Coordination of Laboratory Network Performance of P-Tests 51

10 LC-MS/MS in Routine Analysis Laboratories equipped with LC-MS/MS (Triple Quadrupole) Weiding (Germany): Quattro LC (Waters), API3000 (Applied Biosystems) Cergy (France): API4000 Dublin (USA): API3000 Singapore: Quattro Micro (Waters), API3000, API2000 NRC: different suppliers (Waters, Applied Biosystems, Agilent,...) 52

11 LC-MS/MS Methods for Routine Applications for Banned compounds Compounds, for which other detection techniques are not suitable Methods available or in preparation Chlormequat, Acrylamide Chloramphenicol, Nitrofuran antibiotics, Spinosad residues, Beta-lactam antibiotics, 5-Nitroimidazoles Illegal colorants Multi-residue methods for pesticides and veterinary drugs (in preparation) 53

12 LC-MS/MS Methods for Routine Main purpose Confirmation of analyte absence/presence Criteria in accordance to EU Commission Decision 2002/657/EC Quantitation of analyte Tampering cases: structural information isotopes, molecular weight, functional groups, etc in combination with other techniques (e.g. NMR) 54

13 MS Acquisition Mode SRM Mode (Selected Reaction Monitoring): Q1 Q2 Q3 Q1: mass filter for molecular ion (precursor) Q2: collision cell to generate product ions Q3: mass filter for one product ion per time unit 55

14 Confirmation: EU 2002/257/EC Definition Confirmatory methods for organic residues or contaminants shall provide information on the chemical structure of the analyte. Identification points: Banned compounds: 4 at least 1 precursor and 2 product ions at least 2 SRM traces 56

15 Confirmation: EU 2002/257/EC Criteria Positive response in at least two SRM traces (signal-to-noise: 3) Ion ratios (response ratios) respect of max. permitted tolerances or even more stringent make use of natural isotopic distribution (e.g. 35 Cl/ 37 Cl) Retention time ratio (analyte/internal standard) 57

16 Nitrofuran Antibiotics Added to Honey m/z 335 >291 m/z 236 > µg/kg AMOZ 0.35 µg/kg AOZ Matrices covered Honey, poultry, seafood, milk powder m/z 249 >134 m/z 209 > µg/kg AH 1.1 µg/kg SC References Seu-Ping Khong et al., J. Agric. Food Chem. 2004, 52, P. Mottier et al. J. Chromatogr. A, 1067 (2005)

17 AMOZ Added to Honey m/z 335 >128 Qualifier m/z 335 >262 Qualifier 0.5 µg/kg AMOZ m/z 335 >291 Quantifier m/z 340 >265 m/z 340 >296 Qualifier Quantifier AMOZ-d µg/kg 59

18 Chloramphenicol Added to Milk Powder 100 % 8.13 m/z 328 > Cl-isotope % m/z 326 > Cl-isotope 5 µg/kg CAP-d % m/z 323 > Cl-isotope 100 % % m/z 321 > 257 m/z 321 > Time Cl-isotope 35 Cl-isotope 0.43 µg/kg CAP P. Mottier et al., Food Additives and Contaminants, 2003, 20 (4),

19 Illegal Colorants Added to Chilli Powder 25 µg/kg Sudan Red 7B 100 µg/kg Sudan IV 100 % % m/z 380 > 169 m/z 381 > Other matrices Spice powders e.g. turmeric, curry, paprika Oleoresins e.g. capsicum, paprika Palm oil 50 µg/kg Sudan III % m/z 353 > 156 Synthetic food colours e.g. Sunset Yellow Other colours 50 µg/kg Sudan II 100 % 9.75 m/z 277 > 156 Sudan I, Para Red, Butter Yellow, Toluidine Red Time 61

20 Quality Control: Method Performance Method verification as a part of method implementation (accredited by local authorities) Common controls reagent blanks (contamination) FAPAS samples if available, otherwise spiked sample blanks (recovery and detection limit) solvent blanks (carry-over) instrumental controls (sensitivity) Participation in interlaboratory studies FAPAS, Nestlé P-Test 62

21 LC-MS/MS: Advantages High selectivity in SRM mode low detection limits even in complex food matrices baseline separation of analytes not mandatory Confirmatory analysis one analysis sufficient to prove analyte presence/absence Feasible to analyse difficult molecules e.g. Acrylamide, Chlormequat without derivatisation 63

22 LC-MS/MS: Challenges Broad variety of food matrices raw materials e.g. honey, milk, cereals, vegetables, fruits semi-processed ingredients e.g. chicken meat, milk powder flavourings finished products Methods have to be applicable for varying matrices Challenges due to sample matrix visible and invisible interferences (preferable 3 SRM traces) impact on detection limit (ion suppression) more frequent MS cleaning necessary (e.g. honey, oleoresin) 64

23 LC-MS/MS: Challenges Varying LC-MS/MS instruments different fragmentation pattern e.g. Acrylamide: m/z 27 as product ion with Quattro Micro m/z 44 as product ion with API3000 varying LODs Quantitation internal standards needed for reliable quantitation (otherwise: standard addition, which is time-consuming) Data evaluation tools able to quantify the analyte levels calculate ion ratios 65 identify ion ratios out-of-tolerance range

24 LC-MS/MS: Challenges Banned compounds Nestlé policy: If an ingredient is detected positive for a banned compound, the ingredient will be rejected. with Minimum Required Performance Level (MRPL) LOD at most MRPL set by European Union if LOD lower than MRPL and sample found positive rejection of ingredient without MRPL LOD at least at the same level as commercial and/or food authority laboratories (e.g. Sudan colorants) 66

25 Conclusions LC-MS/MS today a routine technique in Nestlé laboratories a powerful technique to confirm analytes in complex food matrices at low concentration level reliable quantitation possible in complex food matrices by using isotopic-labelled internal standards implementation of applications for multi-residue screening in near future 67

26 Acknowledgment NQAC Singapore Wee Siew-Moi Daniel Tan Chan Wai-Chinn NRC Philippe Guy Pascal Mottier Eric Gremaud Seu-Ping Khong. 68