RI-ERP-FINALACTION-Recommendations

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

For tannin and polyphenol containing samples (chocolate, coffee, cocoa, chestnut, buckwheat, millet and spices) weigh 0.25 g of the homogenized sample, add 0.25 g of skim milk powder and add 2.5 ml of the Cocktail (patented). Close the vial and mix well (vortex) to suspend the sample. Incubate the vial for 40 min at 50°C in the water bath. Let the sample cool down and add 7.5 mL 80% ethanol (3.6.3). Close the vial and shake the vial for 1 h up-side down or by a rotator at room temperature (20-25 °C). Centrifuge the sample (2500 g at least) at room temperature (20-25 °C) for 10 min.; alternatively, let the sample settle down and/or filtrate. Dilute 50 µL supernatant with 500 µL sample diluent (see 3.6.1) in the test tubes (see 3.1.6) and subsequently proceed with 3.8. Sample preparation procedure: surfaces Place 500 µL of sample diluent (see 3.6.1) in the test tube. Swab with the lower end (reaction zone) of a dry dip-stick thoroughly over a sampling area of 10 x 10 cm. Wear gloves during the swabbing procedure. Proceed with 3.8. 3.8.1 General recommendations for good test performance 3.8.1.1 This test should only be carried out by trained laboratory employees. The instructions for use must be strictly followed. No quality guarantee is accepted after expiry of the kit (see expiry label). Do not interchange individual reagents between kits of different lot numbers. 3.8.1.2 Special attention should be directed to the interpretation of positive and negative outcomes (use of evaluation card and control samples). 3.8.1.3 Bring the dip-sticks to room temperature (20-25 °C) before first use (after first use store at room temperature). The dip-sticks are very sensitive to humidity that could turn the test useless. For this reason keep the strips away from humidity. 3.8.1.4 Use also gliadin-free and gliadin-containing samples as test controls (e.g. R7010 for ethanol extraction and R7012 for Cocktail extraction; both products are distributed by R-Biopharm AG, Germany). If the negative assay control sample is evaluated as positive then a contamination of the laboratory or laboratory equipment is likely. 3.8.1.5 It is recommended to compare the extraction efficiency of ethanol with the Cocktail (patented) (R7006) in case of unknown samples Dip-stick testing 3.8.2.1 Place the dip-stick vertically into the test tube filled with the sample extract (prepared as described in 3.7.2 or 3.7.3) or sample diluent after swabbing a surface (3.7.4). The arrow on the dip-stick should point down (see also Fig. 1). Do not immerse the dip-stick beyond the maximum line. 3.8.2.2 Take out the stick after exactly 5 min and evaluate the result using the evaluation card (3.1.9). 3.8.2.3 For documentation and prolonged storage, the upper part of the dip-stick marked with “Gluten” together with the test bands should be cut off. 3.8.2 3.7.4 3.8 Analysis

AOAC Research Institute Expert Review Pan l Use Only

3.9

Evaluation, Reporting and Interpretation of Test Results

3.9.1

Dip-stick evaluation

3.9.1.1 Positive result If two colored bands (test band in red and control band in blue) are visible in the result window (see fig. 1) after 5 min, the sample is positive. In case of testing a

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RIDA®QUICK Gliadin Validation report 2015-01-14