Improved spectrophotometry method for determining proteins by using Coomassie brilliant blue

A Coomassie brilliant blue method and a technology of Coomassie brilliant blue, which are applied in the field of photometry for the determination of proteins by Coomassie brilliant blue, can solve problems such as poor protein stability, and achieve the effect of improving stability and sensitivity

Inactive Publication Date: 2012-06-20
UNIV OF JINAN
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  • Abstract
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  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to overcome the disadvantages of the poor stability of the classic Coomassie brilliant blue method for measuring protein, and provide a photometric method for measuring protein with improved Coomassie brilliant blue, specifically related to a microemulsion based on an improved microemulsion. Sensitive Coomassie Brilliant Blue is a photometric method for the determination of trace proteins, which is characterized by the following steps:

Method used

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  • Improved spectrophotometry method for determining proteins by using Coomassie brilliant blue
  • Improved spectrophotometry method for determining proteins by using Coomassie brilliant blue
  • Improved spectrophotometry method for determining proteins by using Coomassie brilliant blue

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Experimental program
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Effect test

Embodiment 1

[0024] Embodiment 1 (determination of protein in penicillin sodium salt)

[0025] Sample pretreatment: Accurately weigh 0.4800g of penicillin sodium for injection produced by a pharmaceutical company in a beaker, add appropriate amount of water to dissolve, transfer to a 50mL volumetric flask after dissolution, and shake to volume with water.

[0026] According to the mass ratio OP-10: n-butanol: n-heptane: water=2:3:1:90, prepare OP-10 microemulsion, add 2.00mL Coomassie brilliant blue G-250 solution (10.0 μg·mL -1 ), 4.00μLOP-10 microemulsion, 1.00mL sample solution, dilute to volume with water and shake well, let it stand for 25min, and measure the absorbance of the system at 595nm with the reagent blank as a reference. The measurement results are shown in Table 3.

[0027] The analysis result of protein in the penicillin sodium salt of table 3

[0028]

Embodiment 2

[0029] Embodiment 2 (determination of protein in penicillin G potassium industrial salt)

[0030] Sample treatment: Accurately weigh 0.9600g of penicillin G potassium industrial salt produced by a certain company in a beaker, add appropriate amount of water to dissolve, transfer to a 100mL volumetric flask after dissolution, add water to volume and shake well.

[0031] According to the mass ratio OP-10: n-butanol: n-heptane: water=1:4:1:92, OP-10 microemulsion was prepared, and 2.00mL Coomassie Brilliant Blue G-250 solution (10.0 μg·mL -1 ), 4.00μL OP-10 microemulsion, 1.00mL sample solution, shake to a constant volume, let it stand for 15min, and measure the absorbance of the reagent blank at 595nm. The measurement results are shown in Table 4.

[0032] The analysis result of protein in the penicillin G potassium industrial salt sample of table 4

[0033]

Embodiment 3

[0034] Embodiment 3 (determination of protein in milk powder sample)

[0035] Sample treatment: Accurately weigh 0.0313g of commercially available milk powder into a glass mortar, dilute the sample solution to a 100mL volumetric flask, dilute to volume with water and shake well.

[0036] According to the mass ratio OP-10: n-butanol: n-heptane: water=2:3:1:89, prepare OP-10 microemulsion, add 2.00mL Coomassie brilliant blue G-250 solution (10.0 μg·mL-1), 4.00μL OP-10 microemulsion, 2.50mL sample solution, shake at constant volume, let stand for 20min, measure the absorbance of the reagent blank at 595nm. The measurement results are shown in Table 5.

[0037] The analysis result of protein in the milk powder sample of table 5

[0038]

[0039] As can be seen from the measurement results of the above 3 embodiments, the relative standard deviation is less than 5%, indicating that the method precision is higher; the recovery test shows that the recovery rate recorded is all wi...

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Abstract

The invention relates to an improved spectrophotometry method for determining proteins by using Coomassie brilliant blue. For overcoming the shortcoming of poor stability in the conventional Coomassie brilliant blue method for determining proteins, the invention provides a Coomassie brilliant blue method for determining proteins based on increasing stability and sensitivity by adding microemulsion, which employs microemulsion, to effectively improve the stability of a system and the sensitivity of the reaction system. The determination method of the invention is a simple, rapid and quantitative method for determining proteins.

Description

technical field [0001] The invention relates to an improved coomassie brilliant blue photometric method for determining protein, in particular to a microemulsion-stabilized and sensitized coomassie brilliant blue-based photometric method for determining trace proteins, belonging to the fields of biological analysis and food safety. Background technique [0002] Protein is one of the most important basic components of organisms. It is closely related to nutrition, cell structure, enzymes, hormones, viruses, immunity, metabolism, material operation, and genetics. Therefore, the quantitative detection of proteins is widely used in life sciences, clinical medicine, chemical research and Food safety and other fields occupy a very important position. [0003] In recent years, the increasingly in-depth study of protein analysis methods has aroused the interest and widespread concern of scientists. Commonly used quantitative determination methods for protein include: Kjeldahl metho...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/31
Inventor 魏琴杜斌韩颜颜吴丹毛珂霞李贺马洪敏朱宝存范大伟李燕李慧芝罗川南庞雪辉朱沛华赵燕芳蔡燕燕魏东
Owner UNIV OF JINAN
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