Method for detecting content of proteins in chitosan or chitosan salt

Abstract

The invention relates to a method for detecting the content of proteins in chitosan or a chitosan salt. The method comprises the following steps: (1) preparing a Coomassie brilliant blue G-250 test solution; (2) weighing a chitosan or chitosan salt sample, and preparing a sample solution with a concentration of 5 mg/ml; (3) preparing protein standard solutions with different concentrations, addingthe Coomassie brilliant blue G-250 test solution, and drawing an absorbance-concentration curve; and (4) adding the Coomassie brilliant blue G-250 test solution to the sample solution, measuring theabsorbance of the sample solution at 595 nm, and calculating the content of proteins in the sample. A chitosan/chitosan salt system to be detected undergoes ultrasonic treatment under a condition witha hydrochloric acid concentration of below 2% for 15 min, undergoes 100 DEG C water bath treatment, and is stained with the Coomassie brilliant blue G-250 test solution with a specific concentration,and the absorbance at 595 nm is detected, so the accuracy of the protein detection value is significantly improved.

Description

technical field [0001] The invention relates to a method for measuring protein content in chitosan or chitosan salt, belonging to the technical field of protein detection. Background technique [0002] Coomassie Briliant Blue (Coomassie Bri1liant Blue) G-250 is a detection reagent with two hues, which is red in the free state and turns cyan after binding to the protein, and its color depth is directly proportional to the concentration of the protein. Its color has a maximum light absorption at 595nm, which can be measured with a spectrophotometer. Due to its special properties, it is widely used in the field of protein detection. [0003] YY / T 0606.7-2008 describes in detail the method for detecting proteins in chitosan using Coomassie Brilliant Blue (Coomassie Brilliant Blue) G-250. Including the preparation of Coomassie Brilliant Blue G-250 test solution: Weigh 100mg of Coomassie Brilliant Blue G-250 and dissolve it in 50mL of 95% ethanol, then add 100mL of 85% phosphori...

AI Technical Summary

Problems solved by technology

[0008] However, in the actual detection process, the inventor found that there is a certain difference between the above-mentioned detection method and the actual protein content, which cannot meet the high-precision detection of protein in chitosan or chitosan salt

[0009] Although subsequent researchers have improved the above method (Han Ting, Shi Guohua, etc., "Research on the Method for Determination of Protein Content in Chitosan" in "Chinese Journal of Medical Devices", Volume 40, No. 2, 2016), hydrochloric acid was used to replace phosphoric acid The problem of preventing chitosan precipitation has been realized, and the accuracy of protein content determination has been improved, but it still cannot meet the needs of high-precision protein content detection

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