蜜三糖/棉子糖蔗糖/D-半乳糖检测试剂盒Raffinose/

产品货号：K-RAFGL

品牌：Megazyme

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-glucose, sucrose and raffinose, stachyose and verbascose in seeds and seed meals. Based on the measurement of D-glucose on enzymic hydrolysis of raffinose, stachyose and verbascose to D-glucose, D-fructose and D-galactose.

INTRODUCTION:

Grain legumes are an important component of both human and livestock diets. Galactosyl-sucrose oligosaccharides (raffinose, stachyose and verbascose) are major components in many food legumes1, and the antinutritional activity of grain legumes is frequently associated with the presence of these oligosaccharides2. Galactosyl-sucrose oligosaccharides are not hydrolysed in the upper gut due to the absence of α-galactosidase. In the lower intestine they are metabolised by bacterial action, producing methane, hydrogen and carbon dioxide, which lead to flatulence and diarrhoea. Galactosyl-sucrose oligosaccharides are thus a factor limiting the use of grain legumes in monogastric diets3.
Several solvents have been employed for the extraction of galactosyl-sucrose oligosaccharides from legume-seed flours. These are generally water/alcohol mixtures. Before (or concurrent with) extraction, it is vital that endogenous α-galactosidase and invertase are inactivated. This can be achieved by refluxing the flour in ethanol or in an aqueous ethanol mixture before the flour is subjected to aqueous extraction.
Identification and quantification of the extracted galactosyl-sucrose oligosaccharides have been achieved using an array of chromatographic procedures, however many of these methods are, at best, semiquantitative. Chromatographic procedures employing high performance liquid chromatography and low pressure liquid chromatography (using Bio-Gel P2) are quantitative, but can be time consuming, particularly in the area of sample preparation.
It is well known that raffinose, stachyose and verbascose are hydrolysed by α-galactosidase to D-galactose and sucrose. Biochemical kits for the measurement of raffinose are commercially available. The
α-galactosidase used in these kits (from green coffee beans) rapidly hydrolyses raffinose, but acts quite slowly on stachyose and verbascose, and thus does not give complete hydrolysis of these oligosaccharides under the incubation conditions recommended. In contrast, the enzyme used in the current procedure (from Aspergillus niger) readily and rapidly catalyses complete hydrolysis of raffinose, stachyose and verbascose to D-galactose and sucrose.

KITS:

Kits suitable for performing 120 assays of D-glucose, sucrose and raffinose-series oligosaccharides are available from Megazyme. The kits contain the full assay method plus:
Bottle 1: α-Galactosidase suspension (A. niger; 2 mL) in ammonium sulphate.
Stable for > 5 years at 4°C.
Bottle 2: Invertase solution (yeast; 6 mL) containing sodium azide (0.02%) as a preservative.
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
and 4-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: Soy-Flour Reference Sample (containing glucose, sucrose and galactosyl-sucrose oligosaccharides).

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