产品货号：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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