||Enzymatic method for the determination of Sucrose, D-Fructose and D-Glucose. Based on the spectrophotometric measurement of NADPH produced though the reactions, after addition of hexokinase (HK), phosphoglucose isomerase (PGI) and glucose-6-phosphate dehydrogenase (G6PDH). The D-glucose concentration is determined before and after hydrolysis of sucrose by b-fructosidase. The sucrose content is calculated form the difference in D-glucose concentrations before and after hydrolysis by b-fructosidase.
||Sucrose, D-fructose and D-glucose occur widely in plant organisms. In foods, they occur mainly in honey, wine and beer, and a range of solid foodstuffs such as bread and pastries, chocolate and candies. In the wine industry, the addition of sucrose is only allowed in few situations, such as champagne production.
||This rapid and simple specific enzymatic method is used for the simultaneous determination of sucrose, D-fructose and D-glucose in foodstuffs, pharmaceuticals, cosmetics and biological samples.
||Solution 1. Buffer (25 mL, pH 7.6). Stable for 2 years at 4 °C. Solution 2. NADP+ plus ATP and PVP. Stable for 2 years at - 20 °C. Dissolve in 22 mL of distilled water, divide into appropriately sized aliquots and store in PP tubes at -20 °C between use (stable for 2 years) and keep cool during use. Suspension 3. Hexokinase plus glucose-6-P dehydrogenase (4.1 mL). Stable for 2 years at 4 °C. Swirl bottle before use. Suspension 4. Phosphoglucose isomerase (2.25 mL). Stable for 2 years at 4 °C. Swirl bottle before use. Solution 5. D-Fructose/D-glucose standard solution (5 mL, 0.20 mg/mL of each sugar). Stable for 2 years at room temperature. This standard solution can be used when there is some doubt about the method accuracy. Solution 6. Buffer (20 mL, pH 4.6). Stable for 2 years at 4 °C. Suspension 7. β-Fructosidase (2.25 mL). Stable for 2 years at 4 °C. Swirl bottle before use. Add 0.02 ml of Suspension 7 plus 0.180 ml of Solution 6, per assay, to a test tube and homogenise (Solution 6+7). This solution should be prepared for each assay day. Warm Solution 6+7 to 25-30 ºC before use.
|Compatible Sample Types:
||Wine, beer, fruit juices, milk, dietetic foods, bread, jam, honey, ice-creams, fruit and vegetables, pharmaceuticals, cosmetics and biological samples.
|Features & Benefits:
||Rapid reactionStable reagents
||Range: 20-800 mg/LDetection limit: 1.40 mg/L