AOAC OMB Final Action Recommendation (December 2019)-2016.14

FOR EXPERT REVIEW PANEL USE ONLY NOT FOR DISTRIBUTION

Expert Review Panel for Infant Formula and Adult Nutrition Evaluation of Method _____2016.14_____________

Title: Determination of Fructans in Infant Formula and Adult Nutritionals by Anion-Exchange-Chromatography with Pulsed Amperometric Detection after Enzymatic Treatment (AOAC 2016.14): Collaborative Study Authors: Sean Austin, (NR-NIFSAS) and Véronique Spichtig (NR- NIFSAS), Kommer Brunt (Rotating Disc BV, Haren, The Netherlands), Jeroen van Soest and Peter Sanders (Eurofins Carbohydrate Competence Centre, Heerenveen, The Netherlands)

Reviewer Name: Reviewer 2 Summary of Method:

Ion chromatography with PAD detection after solid phase extraction cleanup and enzymatic treatment: Samples are diluted with water and treated with a mixture of sucrase and α-glucanases to hydrolyze sucrose and alpha-glucooligosaccharides to their constituent monosaccharides. The sample is passed through a graphitized carbon SPE column which retains fructans and allows salts and monosaccharides to pass through. Fructans are released from the column using an acetonitrile solution. The released fructans are hydrolyzed with an inulinase mixture, and the released glucose and fructose are analyzed by high performance anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD). The fructan content is calculated by summing the glucose and 0.9 × the fructose content measured. In matrices containing low levels of fructans, a blank correction may be necessary. B. Apparatus Method Scope/Applicability: The m ethod is applicable for the determination of f ructans in infant, adult, and/or pediatric formula (powders, ready-to-feed liquids, and liquid concentrates). General comments about the method: The method is rugged and is an improvement over current AOAC methods (997.08 and 999.03) for the determination of fructans because of the removal of sucrose background and better recoveries of hydrolyzed Fn/Fm fructan portions. Method Clarity: Overall the method is clearly written with detailed instructions, but it might be helpful if some additional information could be provided.

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