AOAC OMB Final Action Recommendation (December 2019)-2016.14

FOR EXPERT REVIEW PANEL USE ONLY NOT FOR DISTRIBUTION

1. If individual enzymes are purchased instead of the Fructan Assay Kit, K-FRUC, could enzyme activity requirements for the individual enzymes be listed in the method, sections C(q) and C(r) ? 2. In sections D(j) and D(k), could more detail be provided about how to adapt the buffer composition if enzymes from another source are used? 3. Can an expiration for reconstituted enzymes be added to sections D(j) and D(k) ? 4. Can an expiration and storage conditions for prepared standard solutions be added to sections D(d), D(e) and D(f) ? 5. In section G , how long should the standards be centrifuged, and could the method include “rcf” requirements? 6. In section H(e) , please clarify if 200 µL of Solution A is transferred to the 1.5 mL microtube when the expected Fructan content is 0.2-0.4 g/100g (powder) or 0.03-0.45 g/100g (RTF). 7. In section H(f), could more information be provided about when it is necessary to add Carrez Clarification? 8. In section H(g) , could some guidelines around “slightly positive pressure” be provided such as maximum drops per minute? If too much pressure is applied, will some of the fructans pass through the SPE column instead of being retained or does IS have the same retention characteristics as fructans and correct for this? 9. For the hydrolysis of sucrose, glucans, and fructans, sections H(e) and H(h) , could more guidelines about how to “adapt these conditions according to enzyme manufacturer recommendations” be provided? 10. In section H(h) , please clarify what are “low amounts of fructans”. 11. Could minimum system suitability requirements be provided? Method Safety Concerns: Method uses sodium azide (when using PA1 column), which is toxic. Pros/Strengths: • The method appears to be rugged given the number and kinds of deviations made to the sample preparation procedure and chromatography during the MLT. • It is not critical to know the type of fructan added to products. Since AOA 2016.14 measures fructose and glucose, it is not necessary to analyze the fructan ingredient or apply factors depending on the fructose to glucose ratio. Cons/Weaknesses • Potential single source enzyme supplier. Supporting Data • General Comment : 1. 14 laboratories from 11 countries participated in the MLT. 2. 12 of the 14 laboratories were able to complete the MLT within the requested timeline. 3. 8 sample matrices were analyzed in duplicate. 4. 3 sets of data were flagged as outliers and removed from the data summary. Method Optimization : NAP to MLT data

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