产品货号：ZZK-K-RAFGL
产品品名：蜜三糖/棉子糖蔗糖/D-半乳糖检测试剂盒
英文品名：Raffinose/Sucrose/D-Glucose Assay Kit
规格型号：120 assays of each per kit
For the measurement of D-Glu cose, sucrose and raffinose, stachyose andverbascose in seeds and seed meals. Based on the measurement ofD-Glu cose on enzymic hydrolysis of raffinose, stachyose and verbascoseto D-Glu cose, D-fructose and D-galactose.
INTRODUCTION:
Grain legumes are an important component of both human and livestockdiets. Galactosyl-sucrose oligosaccharides (raffinose, stachyose andverbascose) are major components in many food legumes1, and theantinutritional activity of grain legumes is frequently associated withthe presence of these oligosaccharides2. Galactosyl-sucroseoligosaccharides are not hydrolysed in the upper gut due to the absenceof Alfa-galactosidase. In the lower intestine they are metabolised bybacterial action, producing methane, hydrogen and carbon dioxide, whichlead to flatulence and diarrhoea. Galactosyl-sucrose oligosaccharidesare thus a factor limiting the use of grain legumes in monogastric diets3.
Several solvents have been employed for the extraction ofgalactosyl-sucrose oligosaccharides from legume-seed flours. These aregenerally water/alcohol mixtures. Before (or concurrent with)extraction, it is vital that endogenous Alfa-galactosidase and invertaseare inactivated. This can be achieved by refluxing the flour in ethanolor in an aqueous ethanol mixture before the flour is subjected toaqueous extraction.
Identification and quantification of the extracted galactosyl-sucroseoligosaccharides have been achieved using an array of chromatographicprocedures, however many of these methods are, at best,semiquantitative. Chromatographic procedures employing high performanceliquid chromatography and low pressure liquid chromatography (usingBio-Gel P2) are quantitative, but can be time consuming, particularly inthe area of sample preparation.
It is well known that raffinose, stachyose and verbascose arehydrolysed by Alfa-galactosidase to D-galactose and sucrose. Biochemicalkits for the measurement of raffinose are commercially available. The
Alfa-galactosidase used in these kits (from green coffee beans) rapidlyhydrolyses raffinose, but acts quite slowly on stachyose and verbascose,and thus does not give complete hydrolysis of these oligosaccharidesunder the incubation conditions recommended. In contrast, the enzymeused in the current procedure (from Aspergillus niger) readily andrapidly catalyses complete hydrolysis of raffinose, stachyose andverbascose to D-galactose and sucrose.
KITS:
Kits suitable for performing 120 assays of D-Glu cose, sucrose andraffinose-series oligosaccharides are available from Megazyme. The kitscontain the full assay method plus:
Bottle 1: Alfa-Galactosidase suspension (A. niger; 2 mL) in ammonium sulphate.
Stable for 大于5 years at 4°C.
Bottle 2: Invertase solution (yeast; 6 mL) containing sodium azide(0.02%) as a preservative.
Stable for 大于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 大于4years 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5 years at room temperature.
Bottle 6: Soy-Flour Reference Sample (containing Glu cose, sucrose andgalactosyl-sucrose oligosaccharides).
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文章号：=2289JP2289