RI-ERP-FINALACTION-Recommendations

OMAMAN-20 C/ In House Validation Report ERP Use Only - March 2015

2.3 Summary of Results

RIDA ® QUICK Gliadin was validated in-house following the AOAC Appendix N for the validation of qualitative methods (1). Wherever necessary, deviation from the Appendix was discussed by the method developer. The method was positively tested with all target compounds (wheat, rye, barley) and negatively tested for all 68 non-target commodities using both claimed extraction methods (60% ethanol or cocktail). The non-target commodities were tested for gluten contamination using the AOAC OMA first action 2012.01 reference method (LoQ of 2.5 mg gliadin/kg). A comprehensive matrix study revealed cut-off values for a naturally contaminated raw corn material of about 1 mg gliadin/kg (ethanol extraction) and 2.2 mg gliadin/kg for an incurred processed snack sample (corn-based; cocktail extracted). By spiking different matrices with WGPAT gliadin, cut-off values between 1 and 2 mg gliadin/kg for unprocessed and 4 mg gliadin/kg for processed matrices were determined. The lower sensitivity of the processed matrices is due to the higher dilution of the extract prior to measurement. As an example for a surface, stainless steel was contaminated with WGPAT gliadin and analyzed by a direct swabbing of the surface with the dip-stick. The outcome was a sensitivity of the assay between 1.5 and 2.0 µg gliadin/100 cm 2 . Ruggedness testing was covered by two collaborative tests under the supervision of AACC. Nevertheless, ruggedness of extraction time, incubation time and temperature was tested, separately. With exception of the incubation time no influence was detected. Increasing the incubation time led to more positive results at a concentration were POD50% was expected. The stability of five independent lots of the RIDA ® QUICK Gliadin was investigated after storage at 4–8°C for up to 24 months. No significant loss of the analytical sensitivity was observed. The lots are comparable in sense of sensitivity.

3

Materials and Methods

3.1

Test Kit Information

3.1.1 3.1.2

Kit Name

RIDA ® QUICK Gliadin

Catalogue Number R7003

Remark : The same dip-stick is also available as R7005 containing buffer vials ready to use for swabbing of surfaces. R7004 contains singled-packaged dip-sticks. The production of the dip- sticks is in all three cases identical. AOAC Research Institute Expert Review Panel Use Only Ordering Information USA R-Biopharm Inc. 870 Vossbrink Drive

Washington, MO 63090 phone (269)-789-3033 fax (269)-789-3070 e-mail s.wesner@r-biopharm.com Germany R-Biopharm AG An der neuen Bergstrasse 17 D-64297 Darmstadt Germany phone +49-(0)6151-8102-0 fax +49-(0)6151-8102-20 e-mail sales@r-biopharm.de

Further information is available at the web site address: www.r-biopharm.com

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RIDA®QUICK Gliadin Validation report 2015-01-14