Method for quantitatively analyzing proteins
A quantitative analysis and protein technology, applied in the field of chemical analysis, can solve problems such as the inability to truly reflect the protein absorbance value, affecting the accuracy of the measured absorbance value, and the limitation of the measurement object, achieving the effect of wide test objects, elimination of influence, and easy experimental operation.
Inactive Publication Date: 2015-06-17
ANHUI POLYTECHNIC UNIV
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However, this detection method needs to prepare a relatively complicated solution. The addition of various additives such as dodecyl salt combination will make the solution turbid, which directly affects the accuracy of the measured absorbance value. The protein component can form the base color with the dye, and the measurement cannot truly reflect the protein absorbance value, and the measurement object of this method is relatively limited
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[0024] The reaction of aromatic primary amine and nitrous acid to form diazonium salt is called diazotization.
[0025] Ar-NH 2 +2HX+NaNO 2 -→Ar-N 2 X+NaX+2H 2 o
[0026] The diazotization reaction should be carried out in strong acid, in fact, nitrous acid acts on ammonium ions. Because nitrous acid is unstable, sodium nitrite and hydrochloric acid or sulfuric acid are usually used to make the nitrous acid generated by the reaction react with primary aramide immediately to avoid the decomposition of nitrous acid. In order for the reaction to proceed smoothly, the primary aromatic amine must first be converted into ammonium cations. Aromatic amines are weakly basic, so diazotization should be carried out in a stronger acid. Some arylamines are very weakly basic and require special methods for diazotization. Diazotization is an exothermic reaction, and diazonium salts are unstable to heat, so it should be carried out under cooling, and the addition rate of sodium nitrite...
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The invention discloses a method for quantitatively analyzing proteins, and belongs to the field of analysis for protein materials. The method includes creating a standard curve of absorbance and tyrosine contents by the aid of ultraviolet-visible spectral characteristics of diazo coupling products of free tyrosine or tyrosine residues in the proteins; quantitatively analyzing the tyrosine; and further deducing contents of compositions of the proteins. The method has the advantages that a test object range is broad, hydrolysis for protein materials is omitted, influence of a hydrolysis procedure to the protein test precision is eliminated, influence of pigments or dye containing N elements on a protein material analysis result is also eliminated, measurement is simple, convenient and feasible, and the accuracy is high.
Description
technical field [0001] The invention belongs to the field of chemical analysis, and in particular relates to a method for quantitative analysis of protein materials, especially for guiding the optimization and characterization of low-damage processing techniques for materials such as real silk, synthetic protein fibers, and down. Background technique [0002] The current standard quantitative analysis method for protein components is the Kjeldahl method based on the N element content test. The biggest problem with this method is that it only recognizes the N element, regardless of its source, which poses a threat to food, materials and other fields. Protein analysis brings a lot of trouble, and even provides opportunities for criminals, such as adding melamine with extremely high N content to milk powder, which has caused extremely bad social problems. At the same time, when protein fibers such as silk, wool, and down are subjected to chemical agents such as inappropriate ac...
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IPC IPC(8): G01N21/31
Inventor 王宗乾李长龙陈维国孙瑞霞
Owner ANHUI POLYTECHNIC UNIV