Method for detecting protein in sodium hyaluronate
A technology of sodium hyaluronate and detection method, which is applied in the direction of material analysis by observing the influence on chemical indicators, and analysis by chemical reaction of materials, etc., can solve the problem of long experimental process time, many detection reagents, high cost, etc. problems, to achieve the effect of shortening the inspection time and reducing the inspection cost
Inactive Publication Date: 2012-06-13
SHANGHAI JINGFENG PHARMA
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Problems solved by technology
The problems of the Lauren method are as follows: 1. It takes a long time to dissolve the sample; it is not easy to mix well during the detection process; 2. After the sample is dissolved, the solution viscosity is high, it is difficult to mix well during the detection process, and the error is large
The problems of the Kewpie method are as follows: 1. There are many detection reagents used and the cost is high; 2. The experimental process takes a long time and the detection efficiency is not high
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Embodiment 4
[0065] The detection method of protein impurity in sodium hyaluronate of the present invention:
[0066] (1) Preparation of standard curve: Accurately weigh about 20.0 mg of bovine serum albumin reference substance, add dilute sodium hydroxide reagent to dissolve and dilute to 100 ml (about 200 μg / ml) to obtain a standard solution, and precisely pipette 1.0 ml of the above standard solution , 2.0ml, 4.0ml, 5.0ml, 10.0ml diluted to 100ml (concentration is about 2μg / ml, 4μg / ml, 8μg / ml, 10μg / ml, 20μg / ml), and accurately pipette 1.0ml of each of the above standard solutions, Precisely pipette 1.0ml of sodium hydroxide reagent into a blank tube (tube 0, make two copies) and place them in 25ml test tubes respectively. Add 1.0ml of reagent E to each of the above tubes, mix them in a vortex shaker immediately, place at room temperature for 10min, add Reagent F3.0ml, immediately mixed in a vortex shaker, placed in a 50°C water bath for 10 minutes, according to the "Ultraviolet Spectrop...
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The invention relates to a method for detecting protein in sodium hyaluronate. The method comprises the following steps of: making a linear regression equation by using an albumin bovine serum standard solution; preparing a sample solution, and measuring an absorbance value of the sample solution; and calculating the content of the protein according to the linear regression equation. The method for making the linear regression equation comprises the following steps of: displaying colors of prepared standard solutions at concentrations of 2 mu g / ml, 4 mu g / ml, 8 mu g / ml, 10 mu g / ml and 20 mu g / ml by using an alkaline copper reagent and Folin-Ciocaileu, and measuring the absorbance at the wavelength of 750 nanometers. By the method for detecting the protein content in the sodium hyaluronate, the detection time is shortened greatly, and the detection cost is reduced; and the method is particularly suitable for detecting the protein content in the sodium hyaluronate in a laboratory.
Description
Technical field: [0001] The invention relates to a method for detecting protein, in particular to a method for detecting protein in sodium hyaluronate. Background technique: [0002] Hyaluronic acid (HA for short) is composed of (1→3)-2-acetylamino-2-deoxy-β-D-glucose-(1→4)-O-β-D-glucuronic acid bis A linear polysaccharide composed of sugar repeating units with the molecular formula (C 14 h 21 NO 11 )n, according to different tissue sources, the molecular weight range is 2×10 5 ~7×10 6 , the number of disaccharide units is 300 to 11,000 pairs. [0003] [0004] Commercial hyaluronic acid is generally in the form of sodium salt, white fibrous or powdery solid, with strong hygroscopicity, soluble in water, insoluble in organic solvents. The macromolecular network structure of sodium hyaluronate passes through the interaction with H 2 O forms hydrogen bonds to bind a large amount of water, and has the functions of forming various matrices, regulating osmotic pressure,...
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IPC IPC(8): G01N21/78
Inventor 薛亮
Owner SHANGHAI JINGFENG PHARMA