Method for detecting thymosin beta 4 content based on high performance liquid chromatograph

A technology of high performance liquid chromatography and determination method is applied in the field of determination of thymosin β4 content based on high performance liquid chromatography, which can solve the problems of low accuracy and sensitivity, time-consuming, small dosage and the like, and achieve reliable detection. method, the effect of accurate measurement

Inactive Publication Date: 2016-08-24
BEIJING NORTHLAND BIOTECH
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Problems solved by technology

The advantage of the Kjeldahl method is that it is not easily affected by other components in the liquid, and the measurement result is accurate, but it is time-consuming; the advantage of the biuret method is that it has good precision and less interfering substances, but its accuracy and sensitivity are low; The advantages of the Lowry method are rapidity and high sensitivity, but it is affected by the specificity of peptides, that is, the color intensity of different peptide drugs is slightly different due to the different contents of tyrosine and tryptophan (Dou Xiaorui et al. (Potency) Determination Method and Its Application. Advan

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  • Method for detecting thymosin beta 4 content based on high performance liquid chromatograph
  • Method for detecting thymosin beta 4 content based on high performance liquid chromatograph
  • Method for detecting thymosin beta 4 content based on high performance liquid chromatograph

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[0036] The present invention will be further described below in conjunction with specific embodiments, and the advantages and characteristics of the present invention will become clearer along with the description. But this example is only normative and does not constitute any limitation on the scope of the invention.

[0037] 1. Experimental materials

[0038] Acetonitrile: purchased from Fisher, grade chromatographically pure

[0039] TFA: purchased from Sigma, the grade is analytically pure

[0040] C18 chromatographic column: purchased from Agilent, the model is Zorbax 300sb-C18

[0041] High-performance liquid chromatography: purchased from Agilent, model 1260.

[0042] 2. Check equipment conditions

[0043] Mobile phase: mobile phase A is water and trifluoroacetic acid, the volume ratio is 999:1

[0044] Mobile phase B is acetonitrile and trifluoroacetic acid, the volume ratio is 999:1

[0045] Chromatographic conditions: flow rate is 1mL / min, 95% A, 5% B solution ...

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Abstract

The invention belongs to the technical field of biological chemistry and provides a high performance liquid chromatograph analysis method for thymosin beta 4 content. In the method, a high performance liquid chromatograph is used, and a gradient elution method is used for content detection. A conventional protein content detecting method for thymosin beta 4 has the defects of being complex in operation, likely to be affected by various factors and the like. Operation is simple, the peak type is good, the separation degree of a main peak and an impurity peak is good, and a simple and reliable analysis method is provided for detecting the thymosin beta 4 content.

Description

(1) Technical field [0001] The invention belongs to the field of biochemistry, and in particular relates to a method for determining the content of thymosin β4 based on high-performance liquid chromatography. (2) Background technology [0002] Thymosin β4 (hereinafter referred to as Tβ4) was isolated from the thymus in the 1980s, and later found to be a small peptide substance widely present in many tissues, and it is an important actin-binding protein. Tβ4 has various biological functions. Studies have shown that Tβ4 acts as a chemical inducer in the wound healing process of myocardium, cornea and skin, regulating the expression of various growth factors and the migration of chemotactic newborn cells. Due to patent factors, currently there are two main companies developing thymosin β4 as a drug, namely RegeneRX Biopharmaceutical Company of the United States, which is in the clinical research stage, and Beijing Northland Biotechnology Co., Ltd., which has applied for clinic...

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Application Information

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IPC IPC(8): G01N30/88
Inventor 许松山马素永聂李亚汤晓闯
Owner BEIJING NORTHLAND BIOTECH
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