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Summary of Method

ER 1 Acceptable

ER 2

It consists of incubation of an aliquot of the sample with thermostable alpha-amylase in pH 5.0 acetate

buffer for 1 hr at 100°C with periodic mixing to gelatinize and partially hydrolyze alpha-glucan.

Amyloglucosidase is added and mixture is incubated at 50°C for 2 h and mixed. After subsequent

addition of water, mixing, clarification, and dilution as needed, free + ezymetically released glucose are

measured using a colorimetric glucose oxidase-peroxidase method. Values from a separate

determination of free glucose are subtracted to give values of enzymatically-released glucose. Dietary

starch = Enzymatically- released glucose multiplied by (162/180) or 0.9 and divided by the as received

sample weight (g) used in the assay.

ER 3

Dietary starch is digested to glucose and the increase in glucose level is used to calculate %dietary

starch. Potential interferences are either accounted for (inherent glucose) or excluded (deter inherent

sucrose digestion and deter maltulose formation).

ER 4

Starch is digested by traditional amylase/amyloglucosidase using gelatinization conditions. Glucose

released is measured colorimetrically with adjustment for free glucose in the sample.

ER 5

Ground or homogenized samples are digested with α-amylase and amyloglucosidate in acetate buffer

to release glucose from dietary starch. The digestate, after optional dilution, is analyzed for its glucose

content. A second sample portion is also assessed for free glucose by treatment with all reagents but

the enzymes. The difference of the two glucose result is used to calculate dietary starch content in the

sample.

ER 6

Sample (containing up to 100mg of starch) is weighed in duplicate. sodium acetate buffer (pH 5.0) is

added to both tubes. Then to one tube alpha-amylase and amylglucosidase are added to hydrolyse the

starch. To the other tube no enzymes are added. Samples are then clarified (centrifugation or

filtration) and diluted. Aliquots from each tube are then taken for analysis of glucose using the glucose

oxidase peroxidase (GOPOD) method, or other suitable validated method for glucose determination.

Glucose determined in the untreated sample is subtracted from the glucose determined in the enzyme-

treated sample. The result is then multiplied by 0.9 to correct for water uptake during hydrolysis to

calculate starch content.

ER 7

good

ER 8

A well performed study. However, the advantages over AOAC Method 996.11 need to be more clearly

identified. A

significant contribution is the application to samples more relevant to the particular study, but some of

the stated

general advantages are not substantiated.

Method Scope/Applicability

ER 1 Animal Feeds and pet foods. 1%-70% starch

ER 2

Animal feedstuffs and pet foods. Limitation in application: The method underestimates dietary starch

in feeds and foods whose antioxidant content is known to exceed 10-20 micromol of hydrophilic

antioxidant (as ascorbic acid) per 0.1 g of test dry matter. The method in the current format may not be

easily applicable to foods/feeds high in phenolic compounds (e.g. beets, red sorghum grain).

ER 3

A wide range of animal and pet feeds were covered in the study. Dry and wet products were included

along with a variety of grains as the base material.

Appendix 1

ERP PROFILE SUMMARIES

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