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A method for separation and analysis of derivatized heparan sulfate disaccharides containing free amino groups

A technology of heparan sulfate and free amino groups, which is applied in the field of separation and analysis of heparan sulfate disaccharides, can solve the problems of decreased sensitivity of mass spectrometry, contamination of ion sources of mass spectrometry, etc., and achieves the expansion of instrument application range, reduction of instrument cost, and great practical value Effect

Active Publication Date: 2021-03-02
FUZHOU UNIV
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Problems solved by technology

Further, patent CN104914205B discloses a kind of N-containing non-substituted glucosamine (GlcNH 3 + ) separation and analysis method of heparan sulfate disaccharides, the mass spectrum peak area correction factor can be used for the quantitative detection of heparan sulfate and heparan disaccharides in actual biological samples, but this method uses amine ion pair reagents for analysis
At present, it is generally believed that ion pair reagents have the risk of contaminating the ion source of the mass spectrometer, which will lead to a decrease in the sensitivity of the mass spectrometer [Yang B, et al. Anal. Biochem., 2011, 415(1):59-66]

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  • A method for separation and analysis of derivatized heparan sulfate disaccharides containing free amino groups
  • A method for separation and analysis of derivatized heparan sulfate disaccharides containing free amino groups
  • A method for separation and analysis of derivatized heparan sulfate disaccharides containing free amino groups

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Embodiment 1

[0036]Use this technical solution to remove N-sulfated heparin (2S, 6S, NH) from self-made heparin derivatives3+-HP) and fully depleted 2,6,N-sulfated heparin (NH3+-HP) The disaccharide component of the sample is analyzed by an example.

[0037]A method for the separation and analysis of derivatized heparan sulfate disaccharides containing free amino groups. The steps are as follows:

[0038](1) Dissolve ammonium acetate in deionized water, adjust the pH to 5.6 with acetic acid, and prepare an ammonium acetate solution with a concentration of 40 mM, which is used as mobile phase A; methanol is used as mobile phase B.

[0039](3) Take 20μg each of the self-made heparin derivative de-N-sulfated heparin and fully de-N-sulfated heparin, and pass the mixture of heparinase Ⅰ, heparinase Ⅱ, and heparinase Ⅲ. After thorough enzymolysis, filter. Add 10 μL of 0.1 mol / L 2-aminoacridone solution (2-aminoacridone dissolved in a 3:17 acetic acid:dimethyl sulfoxide mixed solvent), and react for 20 minutes....

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Abstract

The invention discloses a method for separating and analyzing derivatized heparan sulfate disaccharides containing free amino groups. The functional relationship between the mass of the disaccharides and the peak area detected by mass spectrometry can be used for the quantification of heparin and heparan sulfate disaccharides in actual biological samples detection. The invention can detect heparin and heparan sulfate disaccharides in liquid phase-mass spectrometry analysis, especially the heparan sulfate disaccharides containing free amino groups, and can also be separated by conventional liquid chromatography and analyzed in combination with ultraviolet and fluorescence detectors. This has an important role in the study of clinical drugs heparin, heparan sulfate, heparin derivatives and disaccharide components of various biological samples.

Description

Technical field[0001]The invention belongs to the technical field of drug detection, and specifically relates to a method for separating and analyzing derivatized heparan sulfate disaccharides containing free amino groups.Background technique[0002]Heparin and heparan sulfate are linear glycosaminoglycans. Because heparin has a strong anticoagulant effect, it is widely used in the treatment of thromboembolic diseases, myocardial infarction and other diseases. Different sulfation, acetylation and free amino substitution patterns in heparin form different disaccharide units, which endow heparin and heparan sulfate with numerous biological functions, and are involved in blood vessel formation, blood coagulation, cell adhesion, proliferation, and tumor metastasis. Wait for life activities. Heparin and heparan sulfate are composed of repeating disaccharide units, which are formed by connecting hexuronic acid and glucosamine with 1-4 glycosidic bonds. Among the twelve representative disacc...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/06G01N30/88
CPCG01N30/06G01N30/88
Inventor 魏峥杜佳燕林江慧
Owner FUZHOU UNIV
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