计算溶液所需的质量、体积或浓度。
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在色谱分析、有机合成和交叉偶联反应领域已被 586 篇同行评审文献引用。
Glucose oxidase (GOD, EC 1.1.3.4) exhibits high specificity and can specifically catalyze the oxidation reaction of β-D-glucose. Its systematic name is β-D-glucose:oxygen 1-oxidoreductase. In the absence of catalase, the core of the reaction is that "β-D-glucose is oxidized by O₂ to gluconolactone, with the simultaneous production of H₂O₂".
Application
In clinical analysis, this enzyme can be used for the enzymatic determination of glucose; when maltosaccharides or modified starch are used as substrates, it can also be applied to the detection of amylase activity by coupling with α-glucosidase.
Assay procedure
Principle

The appearance of quinoneimine dye is measured at 555nm by spectrophotometry.
Unit definition
One unit causes the formation of one micromole of hydrogen peroxide(half a micromole of quinoneimine dye)per minute under the conditions described below.
Reagents
A. MES-Na buffer pH5.7:0.1mM [Dissolve 2.13g of 2-(N-morpholino)ethansulfonic acid (MW=213.25) in ca.60ml of H₂O and, after adjusting the pH5.7 with 1N NaOH at 25℃, fill up to 100ml with H₂O] (Stable at 5℃ for one month)
B. Glucose solution: 15% [Dissolve 1.5g of β-D-Glucose and fill up to 10ml with H₂O] for at least 2hrs before assay. (Should prepare fresh)
C. 4-AA solution: 0.5% [50mg of 4-aminoantipyrine (MW=203.25)/10ml of H₂O] (Stable at 5℃ in a brownish bottle for at least one week)
D. TOOS solution: 40mM [132.5mg of N-Ethyl-N-(2-hydroxy-3-sulfopropyl)-m-toluidine (MW=331.36/10ml of H₂O] (Stable at 5℃ in a brownish bottle for at least one week)
E. Peroxidase solution: 500U (purpurogalin unit) /ml of H₂O
F. Enzyme diluent: 10mM MES-Na buffer, pH5.7, containing 0.1% Triton X-100
Procedure
Prepare the following working solution in a brownish bottle and store on ice. (Should be prepared fresh)
| 30ml | Buffer solution | A |
| 6ml | Substrate solution | B |
| 0.3ml | 4-AA solution | C |
| 0.3ml | TOOS solution | D |
| 0.3ml | POD solution | E |
Concentration in assay mixture
| MES buffer | 79mM |
| D-Glucose | 131mM |
| 4-AA | 0.2mM |
| TOOS | 0.3mM |
| POD | ca.4 U /ml |
2. Pipette 3.0ml of working solution into a cuvette (d=1.0 cm) and equilibrate at 37℃ for about 5 minutes.
3. Add 0.1ml of the enzyme solution* and mix by gentle inversion.
4. Record the increase in optical density at 555nm against water for 2 to 3 minutes in a spectrophotometer thermostated at 37℃,and calculate the ∆OD per minute from the initial portion of the curve( △OD test).At the same time,measure the blank rate(∆OD blank)by using the same method as the test except that enzyme diluent (F) is added instead of the enzyme solution.
* Dissolve the enzyme preparation in ice-cold enzyme diluent(F)and dilute to 0.05~0.2U/ml with the same buffer,immediately before assay.
Calcuation
Activity can be calculated by using the following formula:
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| Vt: | Total volume (3.1ml) |
| Vs: | Sample volume (0.1ml) |
| 32.8: | Millimolar extinction coefficient of quinoneimine dye under the assay conditions (cm²/μmol) |
| 1.0: | Light path length (cm) |
| 1/2: | Factor based on the fact that one mole of H₂O₂ produces a half of quinoneimine dye. |
| df: | Dilution factor |
| C: | Enzyme concentration in dissolution (C mg/ml) |
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| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
| 分析证书 | R1505821 | |
| 分析证书 | R1505821 | |
| 分析证书 | R1505821 |
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