Allergen Analysis : Customer Solution Case Studies. ILSI Madrid 2018 Adrian Rogers
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1 Allergen Analysis : Customer Solution Case Studies ILSI Madrid 2018 Adrian Rogers
2 Immunoassay Challenges Every food is different and presents it s own unique challenges with immunoassay analysis If there is any doubt about how a food will perform VALIDATE Knowledge is power, know your food
3 Case Study 1 Peanut Allergen Analysis in Confectionary
4 Peanut Allergen Analysis The Challenge: Large multi-national confectionary company Current ELISA discontinued Very stringent validation procedure Need Kit suppliers full support and complete transparency in communication.
5 Peanut Allergen Analysis Romer s Solution Use AgraQuant PLUS Peanut To be as open and supportive as possible with the customer. To undertake the required validation quickly and thoroughly.
6 Peanut Allergen Analysis Testing Criteria The method must be tested for peanut protein and the final result must be expressed in peanuts. Matrices: ice cream, chocolate and Chocolate Bar Minimum size amount of sample to be analysed: 2g. Spiking concentration: 1 ppm; 5 ppm and 15 ppm peanuts.
7 Peanut Allergen Analysis Testing Criteria Continued Range of peanut quantification should be at least between 1 and 15 mg/kg; Recovery range: %; Intermediate reproducibility < 40% at 95% confidence level Linearity Limit of Detection (LOD) Limit of Quantitation (LOQ) Precision (2 operators over 8 days) Linearity of Dilution
8 Peanut Allergen Analysis Outcomes Romer were able to satisfactorily meet the customers validation criteria. The AgraQuant PLUS Peanut kit was successfully beta tested at 3 manufacturing sites around the world. Romer s kit was adopted as the company s peanut detection kit of choice world wide.
9 Case Study 2 Casein Allergen Analysis in Baby Formula
10 Casein Allergen Analysis The Challenge: Large multinational baby formula company Need to check for casein residue in hydrolysed whey based formula. Complex problematic matrix Issue with low recoveries of spiked casein
11 Casein Allergen Analysis Romer s Solution Use AgraQuant PLUS Casein ELISA Test fully and partially Hydrolysed Formulas for reactivity. Add a known amount of casein (in the form of skimmed milk powder) to each formula to asses recovery. Test at 2 levels, low level near LOQ of assay, High level in the middle of calibration curve.
12 Casein Allergen Analysis Results Casein (ppm) Casein Low Spike No Matrix 1.3 Casein High Spike No Matrix 11.3 % Recovery Formula N/A Formula 1 Low Spike 0.4* 31 Formula 1 High Spike 5.0* 44 Formula 2 <LOQ N/A Formula 2 Low Spike <LOQ (0.6) 43 Formula 2 High Spike Formula 3 <LOQ N/A Formula 3 Low Spike <LOQ (0.6) 48 Formula 3 High Spike *Casein level in unspiked sample subtracted from spiked sample result
13 Casein Allergen Analysis Outcomes Recoveries of High Spike were better then the manufacturer expected. Data to be reviewed by manufacturer. Romer to work with the manufacturer to asses the rest of their infant formula range
14 Case Study 3 Almond and Peanut Allergen Analysis in Seasoning
15 Almond and Peanut Allergen Analysis The Challenge: South American Seasoning producer. Different ELISA kits from different manufactures give different results. How do they know which one is correct?
16 Almond and Peanut Allergen Analysis Romer s Solution Repeat all testing in Romer s ISO17025 compliant laboratory Test using Lateral Flow Device which incorporates a different antibody Test using an orthogonal method : LC-MS/MS that analyses for Almond and Peanut Peptide fragments.
17 Almond and Peanut Allergen Analysis Results Results Chile Results Austria Manufacturer A ELISA kit Romer Labs ELISA kit Manufacturer A ELISA kit Romer Labs ELISA kit Romer Labs LFD LC- MS/MS Almond 19.6 ppm <0.4 ppm 19.3 ppm <0.4 ppm neg neg Peanut 31 ppm 1.1 ppm 9.7 ppm 3 ppm --- neg What would you decide?
18 Almond and Peanut Allergen Analysis Sometimes there is no clear cut answer Need for an analytical tool box approach Customer was grateful that Romer had gone the extra mile to find an answer. Customer reviewed and changed their testing lab based on Romer s findings Assessing almond and peanut allergens using commercially available immunoanalytical kits and LC-MS/MS: a case study Daly et al : Journal of AOAC International Vol. 101, No. 1, 2018
19 Summary Food allergens are extremely diverse No certified reference material no common reference point for ELISA comparison Close collaboration between customer and kit manufacturer necessary Method must be suitable for the application Customised solutions necessary
20
21 Thank you!
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