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Colorimetric analysis method for quickly measuring trypsin

A trypsin and colorimetric analysis technology, applied in the field of trypsin detection, can solve the problems of high detection cost and unsuitability for analysis and detection, and achieve the effects of strong anti-interference ability, simple and fast measurement, and simple preparation method

Active Publication Date: 2013-06-26
SUZHOU INST OF BIOMEDICAL ENG & TECH CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Most of the above methods need the support of large and expensive equipment, and the detection cost is high, so they are not suitable for routine analysis and detection. Therefore, it is necessary to develop a simple, low-cost and sensitive detection method

Method used

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  • Colorimetric analysis method for quickly measuring trypsin
  • Colorimetric analysis method for quickly measuring trypsin
  • Colorimetric analysis method for quickly measuring trypsin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] a. Mix the silver nitrate aqueous solution and the trisodium citrate solution into a 100mL solution, the final concentration is 0.25mM, and conduct electromagnetic stirring;

[0059] b. Add 3 mL of NaBH 4 (10mM) solution was added to the above solution, stirred at normal temperature, and reacted for 30 minutes to obtain water-soluble silver nanoparticles.

[0060] c. 0.2 μM polypeptide containing 3 amino acids was mixed with silver nanoparticles, and after 24 hours, the polypeptides that were not covalently adsorbed on the surface of silver nanoparticles were removed by centrifugation.

[0061] d. Mix the above solution with the solution to be tested containing trypsin, observe the color after 10 minutes of color reaction, and record the absorption spectrum.

[0062] In this example, the color reaction mixed solution gradually changed from yellow to colorless; figure 2 The peak absorbance value of the obtained silver nanoparticles and the peak absorbance value of the...

Embodiment 2

[0064] a. Mix the silver nitrate aqueous solution and the trisodium citrate solution into a 100mL solution, the final concentration is 0.25mM, and conduct electromagnetic stirring;

[0065] b. Add 3 mL of NaBH 4 (10mM) solution was added to the above solution, stirred at normal temperature, and reacted for 30 minutes to obtain water-soluble silver nanoparticles.

[0066] c. 0.2 μM polypeptide containing 3 amino acids was mixed with silver nanoparticles, and after 24 hours, the polypeptides that were not covalently adsorbed on the surface of silver nanoparticles were removed by centrifugation.

[0067] d. Mix the above solution with the solution to be tested containing alkaline phosphatase ALP, observe the color after 10 minutes of color reaction, and record the absorption spectrum.

[0068] In this example, the color of the color reaction mixed solution has almost no change; the peak absorbance value of the obtained reaction product, the peak absorbance value of silver nanopa...

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Abstract

The invention discloses a colorimetric analysis method for quickly measuring trypsin. The method comprises the following steps of: detecting the ultraviolet-visible spectroscopy of silver nanoparticles; carrying out chromogenic reaction between the trypsin and the polypeptide modifying silver nanoparticles; detecting the ultraviolet-visible spectroscopy of the reaction product at the reaction endpoint; calculating the difference value between the light absorption peak value of the silver nanoparticles and the light absorption peak value of the reaction product, and then obtaining the content of the trypsin according to comparison of the difference value with a predetermined standard curve. The detection method provided by the invention has high-sensitivity selective response to the trypsin and the lowest detection concentration reaches 2ng / mL; and the detection method provided by the invention has no need of complex chemical modification or signal marking, and is high in antijamming capability and capable of quickly and accurately realizing the detection of the trypsin, therefore, the detection method is suitable for clinical application, for example diagnosis of diseases such as pancreatitis.

Description

technical field [0001] The invention relates to the technical field of trypsin detection, in particular to a colorimetric analysis method for rapidly measuring trypsin. Background technique [0002] Trypsin is a component of pancreatic juice, a proteolytic enzyme secreted by the pancreas. As an endopeptidase, it can cleave the hydroxyl side of lysine and arginine residues in polypeptide chains. Trypsin is an important protein-digesting enzyme, which can limit the decomposition of precursors of other enzymes such as chymotrypsinogen, carboxypeptidase, and phospholipase, and activate them. It is the most specific protease. It has a wide range of applications in medicine, food, and biochemical research. In addition, the content of trypsin is also an important marker for the clinical diagnosis of pancreatic related diseases. Therefore, the quantitative detection of trypsin plays an important role in the fields of human health and production quality control. [0003] Common ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/31G01N21/33
Inventor 缪鹏韩坤程文播刘涛殷建
Owner SUZHOU INST OF BIOMEDICAL ENG & TECH CHINESE ACADEMY OF SCI
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