An Enzymatic Time-Temperature Indicating System

A technology of time temperature and indicators, applied in thermometers, thermometers with physical/chemical changes, instruments, etc., can solve problems such as unsafe and unsuitable, poor stability, complex system, etc., and achieve safe use, good stability, and detection The effect of high sensitivity

Inactive Publication Date: 2019-03-26
CHINA NAT INST OF STANDARDIZATION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In practical applications, the above-mentioned enzyme-type time-temperature indicators will have the following problems: (1) Most enzyme-type time-temperature indicators need to add additional indicators. , and then add a pH indicator to produce a color change. For example, in 2009, Ning Peng [1] et al. used the hydrolysis reaction of alkaline lipase and glycerol triacetate to reduce the pH value of the system, and mixed phenol red-phenolphthalein-thymolphthalein The solution is used as an indicator to reflect the cumulative change of time-temperature through the color change of the reaction system
What they have in common is that the addition of indicators leads to more complex systems, increasing
And some indicators are toxic, unsafe and not suitable for use in food indicators (2) Because they are all single-enzyme catalytic systems, the stability of the reaction is poor, and the color change of the system is not obvious; for example, in 2013, Ge Lei [6] Research on the time and temperature indicating system based on the principle of enzymatic browning, and develop a new type of time and temperature indicating system based on the enzymatic browning of tyrosinase

Method used

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  • An Enzymatic Time-Temperature Indicating System
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  • An Enzymatic Time-Temperature Indicating System

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] An enzyme-type time-temperature indicator system, the enzyme-type time-temperature indicator system is composed of two parts of two enzymes and two substrates;

[0042] (1) described two kinds of enzymes are glucose oxidase and horseradish peroxidase, and the concentration of two kinds of enzymes is respectively 3.5 μ g / ml and 0.15 μ g / ml;

[0043] (2) The two substrates are glucose and 3,3',5,5'-tetramethylbenzidine, and the concentrations of the two added are 0.3mg / ml and 0.15mg / ml respectively;

[0044] (3) The order of adding the double enzyme and double substrate to the system is: 2000 μl of glucose, 400 μl of 3,3′,5,5′-tetramethylbenzidine, 250 μl of horseradish peroxidase and 500 μl of glucose oxidase;

[0045] (4) According to the above concentration ratio and adding smoothly, add in order to react at different temperatures, and measure the absorbance of the system after reacting for a period of time.

[0046] Practical test: Place the time temperature indicato...

Embodiment 2

[0051] (1) Equipped with buffer solution.

[0052] Accurately weigh 15.605g NaH 2 PO 4 2H 2 O was dissolved in 500ml deionized water to prepare 0.2M phosphate buffered A solution.

[0053] Accurately weigh 35.82g Na 2 HPO 4 12H 2 O was dissolved in 500ml of deionized water to prepare 0.2M slow ethyl phosphate solution.

[0054] Accurately weigh 2g of NaOH and dissolve it in 100ml of deionized water to prepare a 0.5M sodium hydroxide solution and mark it for use.

[0055] Take an appropriate amount of 0.2M phosphate buffer mother solution, add dropwise 0.5M sodium hydroxide solution until the pH of the solution is 5, and mark it for use.

[0056] Mix 84ml of methyl phosphate solution with 16ml of solution B, add dropwise 0.5M sodium hydroxide solution until the pH of the solution is 7.5, and mark it for use.

[0057] (2) Equipped with glucose oxidase and horseradish peroxidase stock solution.

[0058] Precisely weigh 5 mg of glucose oxidase and dissolve it in 5 ml of p...

Embodiment 3

[0072] The difference between embodiment 3 and embodiment 2 is that it reacts at different temperatures, and reacts at a low temperature of 5°C. Observe the color change of the system and the change of the absorbance value of the system. Other condition steps are all identical with embodiment 1.

[0073] From the absorbance curve ( Figure 7 ) and system color change diagram ( Figure 8 ) shows that at low temperature, the absorbance value of the system reaches the maximum after 2h and tends to be stable, indicating that the reaction of the double enzyme is slow at low temperature, indicating that the lower the temperature, the slower the enzyme reaction, and the enzyme reaction has a better relationship with the temperature. dependency. And it can be seen from the color change diagram that the system remains blue within 1 hour, and there will be no other color changes. It shows that the color also has a good dependence on temperature.

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Abstract

The invention discloses an enzyme-type time-temperature indicator system. The enzyme-type time-temperature indicator system is composed of two kinds of enzymes and two kinds of substrates; (1) the two kinds of enzymes are glucose oxidase and horseradish For peroxidase, the concentrations of the two enzymes range from 1 to 5 μg / ml and 0.05 to 0.2 μg / ml respectively, and the two enzymes are added to the system at a volume ratio of 1:1 to 5:1; (2) The two substrates are glucose and 3,3′,5,5′-tetramethylbenzidine, the concentration of which is 0.1~0.4mg / ml and 0.05~0.2mg / ml; (3) double-enzyme double-bottom The order in which the substances were added to the system was as follows: glucose, 3,3′,5,5′-tetramethylbenzidine, horseradish peroxidase and glucose oxidase; React at different temperatures, and measure the absorbance of the system after a period of reaction. The invention is a novel and safe chromogen reagent, which is often used as a chromogen for ELISA and other immunodetection, has the advantages of high detection sensitivity, good stability, etc., and is safe to use.

Description

technical field [0001] The invention relates to an enzyme-type time-temperature indicating system, which belongs to the technical field of food. Background technique [0002] In the process of food circulation, food has gone through stages such as raw material collection, processing, logistics, storage, and shelf sales. In these processes, the temperature and humidity of the food can affect its quality. Among them, for perishable food, the monitoring of its temperature is particularly important. This is because the temperature is difficult to control during the food circulation process. If the temperature is higher than the set temperature for a certain period of time, it will easily cause perishable food to deteriorate, and the expiration date on the traditional food packaging has lost its indication of the effective shelf life of the food at this time. The role of food safety measures cannot be used to warn sellers or consumers, which will easily cause food safety proble...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01K11/16
CPCG01K11/16
Inventor 李强刘鹏段敏戴岳刘文郝佳
Owner CHINA NAT INST OF STANDARDIZATION
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