A kind of detection method of chymotrypsin activity

Abstract

The invention provides a chymotrypsin activity detection method, and relates to the technical field of enzyme activity detection. According to the method, a reaction system containing Ca<2+> and a pre-heating treatment reaction system are used in the detection process, a light absorption value of the reaction systems presents a better linear relation along with changes of time, the activity of chymotrypsin can be stabilized within reaction time, the repeatability of detection results is good, the activity of the chymotrypsin is increased, the lower detection limit is lowered, the sensitivity is high, the stability is good, the cost is low, and all used reagents are safe and free of toxicity and can be widely applied to laboratories and clinical examination.

Description

technical field [0001] The invention relates to the technical field of enzyme activity detection, in particular to a method for detecting chymotrypsin activity. Background technique [0002] Chymotrypsin is a proteolytic enzyme extracted from bovine pancreas. Because of its proteolytic, anticoagulant and anti-inflammatory properties, chymotrypsin is used in the medical field. Chymotrypsin can cut off the carboxy-terminal peptide chain of tyrosine and phenylalanine in the protein peptide chain, so it can debride wounds and eliminate pus, digest concentrated juice and necrotic tissue, and promote the growth of new granulation. [0003] Enzyme activity refers to the ability of an enzyme to catalyze a certain chemical reaction. Enzyme activity can be expressed by the speed at which an enzyme catalyzes a certain chemical reaction under certain conditions. The faster the enzyme catalyzes the reaction, the higher the enzyme activity, and vice versa. Measuring enzyme activity is ...

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Problems solved by technology

Chymotrypsin has been included in the Chinese Pharmacopoeia. The method for detecting the activity of chymotrypsin in the 2010 edition of the Chinese Pharmacopoeia is to hydrolyze the substrate N-acetyl-L-tyrosine ethyl ester at a constant temperature of 25°C to generate The N-acetyl-L-tyrosine of the absorbance value calculates the enzyme activity according to the conversion rate of the absorbance value. This method requires that the constant time of the change rate of the absorbance value within 5 minutes shall not be less than 3 minutes. During the determination process, the reaction system needs Accurately maintain a constant temperature of 25°C, while most domestic UV spectrophotometers on the market do not have a constant temperature adapter, imported spectrophotometers have this device but are expensive, so it is difficult to ensure a constant temperature, for an enzyme-catalyzed reaction, a constant reaction temperature It is a very important parameter. In addition, the stability of the enzyme itself is also crucial to the catalytic reaction, and protease is easy to self-hydrolyze in aqueous solution. Therefore, when using this method to measure the activity of chymotrypsin, the absorbance value often varies with The linearity of the time change is poor, it is difficult to maintain a constant rate of change of 3 minutes, making the measurement imprecise and poor repeatability

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