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Method for detecting alpha-amylase activity by soluble starch resonance scattering spectrometry

A resonance scattering and amylase technology, which is applied in the measurement of scattering properties, color/spectral properties, Raman scattering, etc. Simple equipment and low cost effect

Inactive Publication Date: 2008-12-10
GUANGXI NORMAL UNIV
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AI Technical Summary

Problems solved by technology

Viscometry is rarely used due to poor accuracy and unstable substrates
Microcalorimetry, although relatively simple, is not accurate enough
Although there are many methods for detecting α-amylase activity, the method for detecting α-amylase activity by soluble starch resonance scattering spectroscopy has not been reported yet.

Method used

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  • Method for detecting alpha-amylase activity by soluble starch resonance scattering spectrometry
  • Method for detecting alpha-amylase activity by soluble starch resonance scattering spectrometry
  • Method for detecting alpha-amylase activity by soluble starch resonance scattering spectrometry

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0025] Detect the content of α-amylase in the saliva of 20 people with the method of the present invention:

[0026] 1. Instruments used for reagents:

[0027] Cary Eclipse spectrophotometer (Varian Company, Palo Alto, CA), HH-S 2 Electric constant temperature water bath (Jintan Dadi Automation Instrument Factory), TGL-16G centrifuge (Shanghai Anting Scientific Instrument Factory);

[0028] Soluble starch (China Pharmaceutical Group Shanghai Chemical Reagent Company), α-amylase (165,000U / g Fontes Chemical Reagent Company), polyacrylamide (molecular weight above 5 million, Shanghai Chemical Reagent Purchasing and Supply Station Subpackage Factory); Soluble starch solution: Take 0.25g of soluble starch in a small beaker, add about 5mL of water and mix well, slowly pour it into a 50ml beaker filled with 20ml of boiling water while stirring, and use a washing bottle to transfer all the starch in the small beaker to 50ml In a beaker, continue to stir and heat, boil slowly for 2mi...

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Abstract

The invention discloses a soluble starch resonance scattering spectra method for detecting Alpha-amylase activity. The resonance scattering strength I423nm of Alpha-amylase with known enzyme activity and the resonance scattering strength I0 of reagent blank are utilized for working out Delta I423nm which is equal to the difference between I423nm and I0 and is used as a standard curve. Then, Delta I of Alpha-amylase with unknown concentration is determined. The concentration of Alpha-amylase of an object to be determined is worked out according to a working curve. The soluble starch resonance scattering spectra method has the advantages of simple equipment, convenient operation, easily obtained reagent and low cost. And the method can be finished just through a spectrophotometer. The soluble starch of substrates is nontoxic.

Description

Technical field: [0001] The invention relates to a method for detecting alpha-amylase activity, in particular to a method for detecting alpha-amylase activity by soluble starch resonance scattering spectroscopy. Background technique: [0002] The full name of α-amylase (α-Amylase, AMS) is 1,4-α-D-glucan 4-glucan hydrolase (EC 3.2.1.1), which is a heterogeneous calcium-dependent metalloprotein Enzyme, which acts on the α-1,4 glucosidic bond of starch to generate maltose and glucose. It has been widely used in various fields such as food, fermentation, grain processing, textile, papermaking, medicine, light chemical industry, and oil exploration. It has high clinical application value, and can diagnose diseases according to changes in enzyme activity in the body, such as pancreatic disease and kidney disease leading to increased amylase activity; liver disease can lead to decreased enzyme activity. Therefore, the research on α-amylase is of great significance. At present, f...

Claims

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Application Information

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IPC IPC(8): G01N21/78G01N21/25G01N21/65G01N21/51
Inventor 马纪刘庆业蒋治良
Owner GUANGXI NORMAL UNIV
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