SPSFAM ERP

This result for starch demonstrates the effectiveness of the borohydride reduction step.

5.10 References McCleary, B.V., Murphy, A. and Mugford, D.C., J. AOAC Int. 83, 356-364 (2000).

McCleary, B.V. and Rossiter, P., J.AOAC Int ., 707-717 (2004)

McCleary, B. V., Charmier, L. M. H. McKie, V. A. and Rogowski, A. (unpublished).

6. Validation 6.1 Planning

The purpose of this report is to verify and validate a recently updated fructan assay procedure developed and supplied by Megazyme as detailed in the Fructan Assay Kit booklet (K-FRUC). This method is an extension of AOAC Method 999.03 (Measurement of total fructans in foods; enzymatic/spectrophotometric method). endo -Levanase has been added to the fructanase mixture (which also contains exo - inulinase and endo -inulinase). The presence of endo -levanase facilitates quantitative measurement of levan as well as inulin and branched fructans. 6.2 Performance characteristics Performance characteristics that are investigated within this study are working range, target selectivity (inulin, levan, branched fructans and FOS), specificity [removal of interfering sugars including sucrose, galactosyl-sucrose oligosaccharides (e.g. raffinose and stachyose) starch, maltodextrins and reducing sugars], Limit of Detection (LOD), Limit of Quantitation (LOQ), Trueness ( bias ) and Precision (repeatability and reproducibility). 6.2.1 Working Range The assay follows the Megazyme Fructan Assay Kit (cat. no. K-FRUC) and has a working range of 0.1 to 100% w/w fructan in the sample. For samples containing 0- 10% w/w fructan, 400 mg of sample is extracted with 25 mL of hot water to give a fructan content of 0.1-1.6 mg/mL; equivalent to approximately 3.6-58.2 µ g of sugar in the PAHBAH assay mixture. For samples containing 10-40% w/w fructan, 100 mg of

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