RI-ERP-FINALACTION-Recommendations

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

Abstract

RIDA ® QUICKGliadin is an immuno-chromatographic test for the detection of gluten in foods based on the monoclonal antibody R5. This monoclonal antibody recognizes gluten in wheat, barley and rye with high affinity and specificity. The R5 monoclonal antibody recognizes among others the potentially toxic sequence QQPFP, which occurs repeatedly in the gluten proteins. Raw samples are extracted by a simple ethanol extraction while processed samples are extracted using the Cocktail solution. The dip-stick is also suitable to test for gluten on surfaces by a swabbing procedure. The immuno-chromatographic procedure is finished within 5 minutes. Manufacturer’s in-house study RIDA ® QUICK Gliadin was validated in-house following the AOAC Appendix N (1) for the validation of qualitative methods. 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 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. 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.

AOAC Res arch Institut Expert Review Pan l Use Only

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RIDA®QUICK Gliadin Validation report 2015-01-14