OMB First to Final Action Recommendation Checklist 11-12-18

ERP - GLUTEN ASSAYS: AOAC 2015.05 FAR

2.2 Food matrices for spiking  Cranberry Syrup  Monin Sirup, 18000 Bourges, France  Ingredients: Sugar, water, flavor, citric acid, lemon juice concentrate, color: anthocyans  Sourdough (starter)  Pure sourdough culture on rice basis; gluten‐free  Ernst BÖCKER GmbH & Co. KG, Ringstraße 55‐57, 32427 Minden/Westfalen, Germany 

2.3 Spiking of matrices  Syrup with (hydrolyzed) gliadin 

Both gliadin materials were reconstituted with 60% ethanol at a concentration of 1 mg/ml proteins. 1  ml of syrup was pipetted into a vial and spiked with different amounts of reconstituted PWG or  hydrolyzed gliadin to obtain concentrations of 3, 6, 9, 12, 16, 21, and 30 mg/kg (hydrolyzed) gliadin.  Six extraction replicates per concentration were analyzed according to the test kit insert.   Gluten free sourdough starter with (hydrolyzed) secalin  Both secalin materials were reconstituted with 60% ethanol at a concentration of 1 mg/ml proteins.  One gram of gluten free sourdough (mixture of 50 g water, 50 g rice meal, 5 g sourdough starter) was  spiked with different amounts of reconstituted purified secalin or hydrolyzed secalin to obtain  concentrations of 2.5, 5, 10, and 15 mg/kg (hydrolyzed) secalin. Six extraction replicates per  concentration were analyzed according to the test kit insert.  Fermentation of gluten free sourdough with (hydrolyzed) secalin  Secalin materials were reconstituted with 60% ethanol at a concentration of 1.4 mg/mL. One mL of the  respective secalin solution was pipetted in a Vorwerk Thermomixer followed by one mL of ethanol  which was used to wash the respective secalin solution vial. The ethanolic solution was diluted with 45  mL water and 47 g rice meal was added. After thorough mixing 4.7 g of sourdough starter was added  and mixed. One g of sample were removed at different time points (after 0, 1, 2, 4, 6, and 24 h at room  temperature) and extracted directly with 10 mL 60% ethanol according to the test kit insert. The  preparation of gluten free sourdough was performed according to manufacturer’s recommendation  for starter culture to produce a gluten free sourdough bread. Rice meal and the starter culture were  checked to be gluten‐free.  2.4 Precision data from QC testing  Three commercial beer samples (Becks Gold, Krusovice and Lammsbräu) were tested in frequent  intervals during QC testing of the product RIDASCREEN® Gliadin competitive. Additionally, the gluten‐ free Lammsbräu lager was spiked at concentrations of 5, 15, 45, and 135mg/kg prolamins. The material  for spiking was the calibrator material. These data were used to estimate a reproducibility standard  deviation. Data from 4 lots over a range of up to 24 months tested by 14 different persons were  compiled. 

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