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Method for determining activity of glucose oxidase

A technology of glucose oxidase and oxidation type, which is applied in the field of determination of glucose oxidase activity, can solve the problems of result influence and cumbersome operation, and achieve the effects of short reaction time, accurate detection results and high stability

Pending Publication Date: 2020-07-10
HEC PHARM CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although spectrophotometry is more sensitive, its results are easily affected by substances such as insoluble particles and pigments in the sample, and the operation is more cumbersome

Method used

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  • Method for determining activity of glucose oxidase
  • Method for determining activity of glucose oxidase
  • Method for determining activity of glucose oxidase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] The specific detection conditions are as follows, and the rest of the operations are the same as the general method above:

[0069] Chromatographic column: Welch-C30 (4.6*250mm 5um); wavelength: 525nm; column temperature: 28°C; flow rate: 1.0ml / min; running time: 40min; mobile phase: A: 86mM sodium acetate solution (pH5.5); B: Methanol; the elution gradient is shown in Table 2.

[0070] Table 2: Elution Gradient

[0071] time (min) A B 0 45％ 55％ 20 15％ 85％ 26 15％ 85％ 26.1 45％ 55％ 40 45％ 55％

[0072] The detection results of the standard product enzyme activity and peak area are shown in Table 3, with the peak area of ​​the product as the abscissa, and the enzyme activity (U / mL) of the standard product as the ordinate, a standard curve is made, see figure 1 , Substituting the peak area obtained after the HPLC detection of the test solution into the standard curve, the enzyme activity of the corresponding test solution can b...

Embodiment 2

[0081] In this example, the inventors further adjusted the column temperature, sodium acetate solution concentration and pH, in order to optimize or obtain a reasonable range for chromatographic detection,

[0082] Under the situation of adjusting column temperature, sodium acetate solution concentration, pH, when the inventor evaluates the influence of single factor chromatographic condition on detection result, fixed all the other detection conditions except the single factor chromatographic condition to be evaluated are the same as embodiment 1, adjusted Single factor chromatographic conditions to be evaluated.

[0083] During the experiment, one injection of standard solution, sample solution and blank solution was injected under each condition, the chromatogram was recorded, the area of ​​the main peak was reported, and the enzyme activity was calculated. The results are shown in Table 5, when the above conditions were changed, the deviation of the measured value of the e...

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Abstract

The invention provides a method for determining the content of pink oxidized o-dianisidine and a method for determining the activity of glucose oxidase. The method for determining the content of pinkoxidized o-dianisidine comprises the following steps: (1), carrying out high performance liquid chromatography detection on pink oxidized o-dianisidine to be detected; and (2), determining the peak area of the pink oxidized o-dianisidine to be detected based on a chromatographic detection result, so as to obtain the content of the pink oxidized o-dianisidine to be detected. The method is simple and easy to implement, short in consumed time, low in cost, high in stability, small in detection error and high in sensitivity.

Description

technical field [0001] The invention relates to the field of detection, in particular, the invention relates to a method for measuring the activity of glucose oxidase. Background technique [0002] Glucose oxidase (Glucose oxidase, E.C.1.1.3.4, referred to as GOD) is an aerobic dehydrogenase that can oxidize β-D-glucose to generate gluconic acid and hydrogen peroxide under aerobic conditions. It is used in food additives, It has a wide range of applications in animal feed additives, biochemical detection and biosensors. [0003] At present, the main detection methods of glucose oxidase activity include barometric method, titration method, electrochemical method and spectrophotometric method. The principle of the air pressure method is that glucose oxidase catalyzes the reaction between glucose and oxygen, and can consume part of the oxygen in the reaction. By measuring the oxygen consumption within a certain period of time, the enzyme activity can be calculated. This metho...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06
CPCG01N30/02G01N30/06
Inventor 屈代鑫刘权吴广进谢文平王小龙赵裕栋徐军魏汉林李峰冯玉枚
Owner HEC PHARM CO LTD
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