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Page 16. Point (b) is missing.

Page 16 – Purity and source of alpha-amylase and AMG.

Detailed specifications on the source of alpha-amylase are given. However, it must be remembered

that these enzymes are made for industrial use. There may be variation from batch to batch in

contaminants important in an analytical procedure but of no consequence in the intended industrial

application.

Industrial AMG cannot be used in analytical procedures because it contains glucose, but more

importantly, because they contain contaminating activities that interfere with starch determination in

plant samples. As far as I am aware, the Megazyme purified AMG (E-AMGDF) is the only AMG pure

enough to use in such assays other than pure AMG, which is too costly to use in such assays.

Page 17, line 336. Change “amylase” to “amyloglucosidase”

Page 17 (e). A statement should be made about the required purity of glucose oxidase and peroxidase.

Page 17, line 378. Phenol is generally not used in glucose determination reagents because it is

carcinogenic and also is not very stable. The chemical most commonly used in its place is

p

-

hydroxybenzoic acid.

ER 9

For me the term "dietary starch" is new, especailly in connection with animal feed and pet food.

Fromenergetic viewpoint, I can agree to include maltodextrins, glycogen fromanimal and microbial

origin in the new term dietary starch. However I have problems what to do with the4 different types of

resistant starhes, the RS1, RS2, RS3 and RS4. Starch incubation with alpha-amylase at 100 C will

hydrolyse the RS1, RS2 and RS4 resistant starch but certainly not the RS3 resistant starch, the so-called

retrograded starch.

Different animals have different intestinal tracks, for example, pets, pigs, cows some can digest

resistant starches, others not. So the content of dietary starch in a feed sample depends also on which

kind of animal consumes the feed.The for digestion available "dietary starch" in one sample containing

resistant starch categories RS1+RS2+RS3+RS4 is most likely different for pets (originally carnivores and

less capable to digest native starches), pigs, cows.

Method Clarity

ER 1 Positive feedback from collaborators

ER 2

Good with the exception how the limitation of the method in application to matrices containing

hydrophilic antioxidant contents/activity exceeding 10-20 micromol as ascorbic acid) per 0.1 g of test

dry matter.

ER 3

Easy to read. No issues.

ER 4

Well written and understandable

ERP PROFILE SUMMARIES

273