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Method for analyzing mucopolysaccharide with glass-carrying electrophoresis method

A technology of mucopolysaccharide and electrophoresis, which is applied in the direction of material analysis, material analysis, and measuring devices through electromagnetic means. It can solve the problems of limited agarose gel electrophoresis samples, difficulty in meeting large-scale analysis of pharmaceutical factories, and spot fading. , to achieve the effect of low production environment and equipment requirements, easy large-scale analysis, and simple operation

Active Publication Date: 2011-01-05
SHENZHEN TECHDOW PHARM CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the current electrophoresis method has certain defects. First, polysaccharides are easy to escape from the gel plate during dyeing and decolorization after electrophoresis, which seriously affects the analysis results; secondly, the electrophoresis results are not easy to preserve. The polysaccharide spots will fade due to the diffusion of polysaccharides; finally, the amount of sample loaded by agarose gel electrophoresis is limited, which is difficult to meet the needs of large-scale analysis of pharmaceutical companies

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Prepare several clean and dry glass plates (7.8cm×6.0cm, 1.5mm thick), wrap them tightly around the perimeter of the glass plates with hard tape, place them on a horizontal table, and set the format comb horizontally at a position 5mm from the lower end. Dissolve agarose in boiling water in barium acetate-acetic acid buffer solution with a pH of 5.8 and a concentration of 0.04 mol / L to make the agar concentration 0.5%, and pour it on a glass plate with a thickness of 2mm when cooled to about 50°C. After solidification, remove the format comb, and tear off the adhesive tape to become the gel plate for electrophoresis.

[0018] Put the two electrophoresis plates side by side into the electrophoresis tank, take 3 μL of the mucopolysaccharide solution with a concentration of 1 mg / ml and put it on the sample hole, and first add it to the barium acetate-acetic acid buffer solution with a pH value of 5.8 and a concentration of 0.04 mol / L at 6.4 mA / cm gel width steady current e...

Embodiment 2

[0021] Prepare several clean and dry glass plates (7.8cm×6.0cm, 1.5mm thick), wrap them tightly around the perimeter of the glass plates with hard tape, place them on a horizontal table, and set the format comb horizontally at a position 5mm from the lower end. Dissolve agarose in boiling water in pH 5.8, 0.04mol / L barium acetate-acetic acid buffer solution to make the agar concentration 0.5%, and pour it on a glass plate with a thickness of 3mm when cooled to about 50°C. After solidification, remove the format comb, and tear off the adhesive tape to become the gel plate for electrophoresis.

[0022] Put the two electrophoresis plates side by side into the electrophoresis tank, take 4 μL of the mucopolysaccharide solution with a concentration of 1 mg / ml and put it on the sampling hole, and first add it to the barium acetate-acetic acid buffer solution with a pH value of 5.8 and a concentration of 0.04 mol / L. 5mA / cm gel width steady flow electrophoresis for 1 hour, and then in ...

Embodiment 3

[0025] Prepare several clean and dry glass plates (7.8cm×6.0cm, 1.5mm thick), wrap them tightly around the perimeter of the glass plates with hard tape, place them on a horizontal table, and set the format comb horizontally at a position 5mm from the lower end. Dissolve agarose in boiling water in a barium acetate-acetic acid buffer solution with a pH value of 5.8 and a concentration of 0.04 mol / L to make the agar concentration 0.8%, and pour it on a glass plate with a thickness of 1mm when cooled to about 50°C. After solidification, remove the format comb, and tear off the adhesive tape to become the gel plate for electrophoresis.

[0026] Put the two electrophoresis plates side by side into the electrophoresis tank, take 5 μL of the mucopolysaccharide solution with a concentration of 1 mg / ml and put it on the sample hole, and first add it to the barium acetate-acetic acid buffer solution with a pH value of 5.8 and a concentration of 0.04 mol / L. 6mA / cm gel width steady flow e...

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Abstract

The invention discloses a method for analyzing mucopolysaccharide with glass-carrying electrophoresis method, comprising the following steps: paving agarose gel on a glass board to form an electrophoresis glue board; arranging the whole glass-carrying board into a proper electrophoresis tank; spotting the sample; performing electrophoresis; taking out the glass-carrying board; drying at the temperature of 60-80 DEG C; dying with toluidine blue; decolouring with tapping water; drying at the temperature of 60-80 DEG C; and finally obtaining the electrophoresis result. The method can store the electrophoresis board for long term; the individual feeding amount is enhanced to 72 from 12-24 in the flat board electrophoresis, which is convenient to perform large-scale analysis; the invention caneffectively separate mucopolysaccharide matters such as heparin, dermatan sulfate, chondroitin sulfate and heparan sulfate and helps to accurately have quantitative determination for the mucopolysaccharide.

Description

technical field [0001] The invention relates to a method for analyzing mucopolysaccharides, in particular to a method for analyzing mucopolysaccharides by glass-mounted electrophoresis. Background technique [0002] Mucopolysaccharides (Mucopolysaccharides) are a class of extracellular biopolymers, which are the sugar chain parts of proteoglycan (Proteoglycan) molecules in animals. Mucopolysaccharides are important biopolymer compounds, which have been confirmed to have a variety of pharmacological activities, including anticoagulant, hypolipidemic, antiviral, antitumor, and antiradiation effects. Representative mucopolysaccharides include heparin, heparan sulfate, dermatan sulfate, and chondroitin sulfate, which are long chains of sulfated and acetylated sugar amines and uronic acid disaccharide repeating units. [0003] These mucopolysaccharides have similar structures and are all discretely distributed sugar chain combinations, which increases the difficulty of analysis....

Claims

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Application Information

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IPC IPC(8): G01N27/447
Inventor 史绍鹏徐归来李建科
Owner SHENZHEN TECHDOW PHARM CO LTD
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