Measurement of Total Starch in Cereal Products by Amyloglucosidase-α-Amylase Method: Collaborative Study

Barry V. McCleary, T. S. Gibson, David C Mugford, O. M. Lukow, David S. Jackson, E Rabe, Nikita Patel, Phil Williams, J. A. Gelroth, Mary Ellen Camire,

Journal of AOAC International · 1997 · 494 citations · 0 references

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TL;DR

The study evaluated the accuracy and reliability of an enzyme assay kit for measuring total starch across various cereal grains and products. The assay involved incubating flour at 95 °C with thermostable α‑amylase, adjusting pH, treating with purified amyloglucosidase to convert dextrins to glucose, and quantifying glucose via a glucose‑oxidase/peroxidase reaction; 32 collaborators analyzed 16 blinded samples of diverse cereal products, with some samples also tested using a DMSO‑cooking step. Repeatability and reproducibility relative standard deviations ranged from 2.1–3.9 % and 2.9–5.7 %, respectively, with the highest RSDr of 5.7 % observed for high‑amylose maize starch.

Abstract

Abstract American Association of Cereal Chem- ists/AOAC collaborative study was conducted to evaluate the accuracy and reliability of an enzyme assay kit procedure for measurement of total starch in a range of cereal grains and products. The flour sample is incubated at 95°C with thermostable α-amylase to catalyze the hydrolysis of starch to maltodextrins, the pH of the slurry is adjusted, and the slurry is treated with a highly purified amyloglucosidase to quantitatively hydrolyze the dextrins to glucose. Glucose is measured with glucose oxidase-peroxidase reagent. Thirty-two collaborators were sent 16 homogeneous test samples as 8 blind duplicates. These samples included chicken feed pellets, white bread, green peas, high- amylose maize starch, white wheat flour, wheat starch, oat bran, and spaghetti. All samples were analyzed by the standard procedure as detailed above; 4 samples (high-amylose maize starch and wheat starch) were also analyzed by a method that requires the samples to be cooked first in dimethyl sulfoxide (DMSO). Relative standard deviations for repeatability (RSDr) ranged from 2.1 to 3.9%, and relative standard deviations for reproducibility (RSDr) ranged from 2.9 to 5.7%. The RSDr value for high amylose maize starch analyzed by the standard (non-DMSO) procedure was 5.7%; the value