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Method for determining trypsin activity by using silver nanoparticles

A technology of silver nanoparticles and trypsin, which is applied in the field of enzyme activity detection, can solve problems such as time-consuming, cumbersome steps, and complicated processes, and achieve the effects of fast and accurate inspection, simple preparation, and good reproducibility

Inactive Publication Date: 2020-06-19
NANJING UNIV OF SCI & TECH
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  • Abstract
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  • Claims
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Problems solved by technology

However, the above method has limitations such as complex process, cumbersome steps, and long time consumption.

Method used

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  • Method for determining trypsin activity by using silver nanoparticles
  • Method for determining trypsin activity by using silver nanoparticles
  • Method for determining trypsin activity by using silver nanoparticles

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Embodiment 1

[0020] The method for measuring trypsin activity using silver nanoparticles, the process is as follows figure 1 As shown, it specifically includes the following steps:

[0021] (1) Preparation of 30nm silver nanoparticles. First, a 4 nm silver seed solution was prepared. 20 mL of 1% citrate solution and 75 mL of deionized water were added to a round bottom flask, and the mixture was heated to boiling for 15 minutes. After that, 1.7 mL of 1% silver nitrate solution and 2 mL of 0.1% freshly prepared sodium borohydride solution were sequentially added to the mixture. The reaction solution was kept boiling with stirring for 1 hour and then cooled to room temperature. Water was then added to the solution to make up to 100 mL. The resulting solution was used as the starting seed solution. Mix 2mL of 1% citrate solution with 80mL of water, and boil for a period of time under constant temperature conditions. Next, 10.0 mL of the above seed solution and 1.7 mL of 1% silver nitrat...

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Abstract

The invention discloses a method for determining trypsin activity by using silver nanoparticles. The method comprises the steps of mixing silver nanoparticle sol with a trypsin solution to be detected; adding casein as a substrate to react, taking trichloroacetic acid as a terminating reactant; after the reaction is finished, carrying out centrifugal separation, measuring the absorbance of supernatant by using a spectrophotometer; and calculating the activity of trypsin to be measured according to the linear relationship between the activity of trypsin and the absorbance. According to the method, the trypsin activity is determined by utilizing the silver nanoparticles; the method has the advantages of simple preparation, no need of extraction and color development, simple steps and the like; the time cost and the experiment cost are greatly reduced; the reproducibility is good; the detection is rapid and accurate; and the actual detection work requirements are met.

Description

technical field [0001] The invention relates to a method for measuring trypsin activity by using silver nanoparticles, belonging to the field of enzyme activity detection. Background technique [0002] Trypsin is a proteolytic enzyme that is very important to the human body. Its relative molecular mass is 23,000, and it consists of 223 amino acid residues. Its protein molecule contains 4 tryptophan residues and 6 pairs of disulfide bonds. In humans, trypsin is one of the most important digestive proteases secreted by the pancreas into the small intestine. It also exists in esophagus, stomach, liver, lung, intravascular epithelial cells, spleen leukocytes and even skin cells, and is widely expressed in other tissues of the human body. [0003] The existing methods for measuring trypsin activity are relatively complicated and difficult to operate in experimental applications. Therefore, how to efficiently measure trypsin activity has become a research hotspot today. Liu Junh...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/31G01N1/28
CPCG01N21/3103G01N1/28
Inventor 陆锐刘敏王连军李健生沈锦优孙秀云
Owner NANJING UNIV OF SCI & TECH
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