allergen analysis : customer solution case...
TRANSCRIPT
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
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.
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.
Peanut Allergen Analysis
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.
Peanut Allergen Analysis Testing Criteria Continued Range of peanut quantification should be at least
between 1 and 15 mg/kg; Recovery range: 60 – 130 %; 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
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.
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
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.
Casein Allergen Analysis
Results Casein (ppm) % Recovery
Casein Low Spike No Matrix 1.3 Casein High Spike No Matrix 11.3 Formula 1 2.6 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 9.7 86 Formula 3 <LOQ N/A Formula 3 Low Spike <LOQ (0.6) 48 Formula 3 High Spike 8.0 71
*Casein level in unspiked sample subtracted from spiked sample result
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
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?
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.
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
Almond and Peanut Allergen Analysis Results
What would you decide ?
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
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
Summary