产品货号:ZZK-K-AMYL
产品品名:直链支链淀粉检测试剂盒
英文品名:Amylose/Amylopectin assay Kit
规格型号:100 assays per kit
For the measurement of amylose/amylopectin ratio and content in cerealstarches and flours. Based on a Con A precipitation procedure.
INTRODUCTION:
Many of the properties of cereal starches that determine theirsuitability for particular end-uses are dependent upon theiramylose/amylopectin ratios. These properties include gelatinisation andgelation characteristics, solubility, the formation of resistant starch,and, for rice, the cooking and textural characteristics of wholegrains1-5. Thus, the measurement of the amylose content of starches isan important quality parameter for starch processing.
Amylose is most commonly determined in cereal starches by thepotentiometric, ampometric or colourimetric measurement of the iodinebinding capacity of the amylose with the resultant formation ofamylose-iodine inclusion complexes6-10. However, these methods aresubject to uncertainties. Amylopectin-iodine complexes also form, andthese reduce the concentration of free iodine measured by thenon-colourimetric methods and may absorb at similar wavelengths toamylose-iodine complexes in colourimetric methods. These complexes leadto an overestimation of the amylose, requiring corrections to beapplied. Many of the other problems experienced in the use of thesemethods are detailed by Gibson et al.11.
The specific formation of amylopectin complexes with the lectinconcanavalin A (Con A) offers an alternative approach to amylosemeasurement in starches, that is not subject to theseuncertainties12,13. Under defined conditions of pH, temperature andionic strength, Con A specifically complexes branched polysaccharidesbased on α-D-glucopyranosyl or α-D-mannopyranosyl units at multiplenon-reducing end-groups with the formation of a precipitate. Thus, Con Aeffectively complexes the amylopectin component of starch but not theprimarily linear amylose component.
The procedure described in this booklet13 is a modification of a Con Amethod developed by Yun and Matheson (1990)13. It uses an ethanolpretreatment step to remove lipids prior to analysis [modified fromMorrison and Laignelet (1983)7].
KITS:
Kits suitable for performing 100 assays are available from Megazyme.The kits contain the full assay method plus:
Bottle 1: Freeze dried Con A.
Stable for > 5 years at -20°C.
Bottle 2: Amyloglucosidase [200 U on p-nitrophenyl
β-maltoside (i.e. 3300 U on starch at pH 4.5 at 40°C)] plus fungalα-amylase (500 U on Ceralpha Reagent at pH 5.0 and 40°C), 2 mL.
Stable for > 5 years at 4°C.
Bottle 3: GOPOD Reagent Buffer. Buffer (48 mL,
pH 7.4), p-hydroxybenzoic acid and sodium azide
(0.04% w/v).
Stable for > 4 years at 4°C.
Bottle 4: GOPOD Reagent Enzymes. Glucose oxidase plus peroxidase and4-aminoantipyrine. Freeze- dried powder.
Stable for > 5 years at -20°C.
Bottle 5: D-Glucose standard solution (5 mL, 1.0 mg/mL) in 0.2% (w/v)benzoic acid.
Stable for > 5 years at room temperature.
Bottle 6: Starch reference sample (with specified content of amylose).
Stable for > 5 years at room temperature.
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