K-TSTA，淀粉总量检测试剂盒，Total Starch (AA/AMG) Assay Kit

K-TSTA，淀粉总量检测试剂盒，Total Starch (AA/AMG) Assay Kit

产品识别号：K-TSTA

产品品名：淀粉总量检测试剂盒

英文品名：Total Starch (AA/AMG) Assay Kit

规格型号：100 assays per kit

AOAC Method 996.11 AACC Method 76-13.01 RACI Standard Method for themeasurement of total starch in cereal flours and food products. AACC andAOAC approved method. This kit now contains an improved α-amylase thatallows the amylase incubations to be performed at pH 5.0 (as well as pH 7.0).

INTRODUCTION:

Starch determination methods are broadly grouped into acid

hydrolysis or enzymic procedures.1 Acid hydrolysis procedures

can only be applied to pure starch samples and thus have limited

application. Enzymic procedures vary in pre-treatment steps,2 starch

gelatinisation, liquefaction and dextrinisation, hydrolysis of dextrins to

Glu cose and Glu cose measurement3. AACC Method 76-114 specifies

starch gelatinisation under aqueous conditions in an autoclave,

followed by starch conversion to Glu cose with amyloglucosidase and

Glu cose measurement. AACC Method 76-11 underestimates starch

content in a range of samples and materials, including high amylose

maize starches and many processed cereal products. Most methods in

use today incorporate treatment with thermostable α-amylase either

during or immediately following the starch gelatinisation step.5,6 For

samples which are difficult to gelatinise (such as high amylose maize

starch) solvents such as sodium hydroxide or dimethyl sulphoxide

(DMSO)7,8 have been employed. In a procedure to ensure complete

solubilisation of starch in dietary fibre determination, Englyst and

Cummings (1988)7 included treatment with the starch debranching

enzyme, pullulanase.

To satisfy the need for an extremely simple, yet quantitative and

reliable, procedure for the measurement of total starch Megazyme

produces, and offers, a total starch assay kit based on the use of

thermostable α-amylase and amyloglucosidase (McCleary et al.9). This

method has been adopted by AOAC (Official Method 996.11) and

AACC (Method 76.13).

More recently, thermostable α-amylases that are active and stable

at lower pH values have become available. Consequently, we have

updated our total starch methodology to incorporate such an

enzyme10. The major advantage of this improvement is to allow both

the thermostable α-amylase and amyloglucosidase incubation steps

to be performed at the same pH (pH 5.0) which, in turn, simplifies

the assay and minimises the possibility of production of maltulose

(4-α-glucopyranosyl-D-fructose)11, which is resistant to hydrolysis by

amyloglucosidase and α-amylase.

The Megazyme total starch analysis procedure allows the

measurement of total starch in a wide range of food, feed, plant and

cereal products (natural or processed). For most samples (e.g. wheat

flour), starch is completely solubilized on incubating the sample at

approx. 100°C in the presence of thermostable α-amylase. Samples

containing high levels of resistant starch (e.g. high amylose maize

starch), require pre-dissolution in cold 2 M KOH or hot DMSO. For

samples containing soluble starch or maltodextrins, cooking with

thermostable α-amylase is not required.

KITS:

Kits suitable for performing 100 determinations of total starch are

available from Megazyme. The kits contain the full assay method plus:

Bottle 1: Thermostable α-amylase (10 mL, 3,000 U/mL

on Ceralpha reagent* at pH 6.5 and 40°C or 1600

U/mL on Ceralpha reagent at pH 5.0 and 40°C).

Stable for 大于 4 years at 4°C.

Bottle 2: Amyloglucosidase (10 mL, 3300 U/mL on soluble

starch (or 200 U/mL on p-nitrophenyl β-maltoside*)

at pH 4.5 and 40°C.

Stable for 大于 4 years at 4°C.

*Full assay procedure is available at “www.megazyme.com”.

Bottle 3: GOPOD Reagent Buffer. Buffer (48 mL,

pH 7.4), p-hydroxybenzoic acid and sodium azide

(0.4 % w/v).

Stable for 大于 4 years at 4°C.

Bottle 4: GOPOD Reagent Enzymes. Glu cose

oxidase plus peroxidase and 4-aminoantipyrine.

Freeze-dried powder.

Stable for 大于 5 years at -20°C.

Bottle 5: D-Glu cose standard solution (5 mL, 1.0 mg/mL) in

0.2 % (w/v) benzoic acid.

Stable for 大于 4 years at room temperature.

Bottle 6: Standardised regular maize starch control. Starch

content shown on vial label.

Stable for 大于 4 years at room temperature.

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