Hirudin extraction separation technique

A technology of leech extract and hirudin, which is applied in the directions of leech inhibitors, protease inhibitors, specific peptides, etc., can solve the problems of insufficient magazine removal, excessive use of organic solvents, complicated extraction methods, etc., and reduce the cost of extraction and preparation. , The effect of reducing the use of organic solvents, reducing product cost and operation difficulty

Inactive Publication Date: 2015-07-08
JINGJIANG ZHIGAO BIOLOGICAL SCI & TECH CO LTD
View PDF1 Cites 8 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This extraction method is complex, difficult to operate, the removal of impurities is not thorough enough, the extraction efficiency is low, and more organic solvents are used, causing great environmental pollution.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0027] 1L of leech extraction stock solution is filtered through a ceramic filter membrane, and the specification of the ceramic filter membrane is 100KDa molecular weight cut-off, and the distilled water of 30% volume of the ceramic filter membrane filtrate is used for cleaning to obtain a total of 1.03L of filtrate and cleaning solution; Pass through a 30KDa ceramic filter membrane, and wash with distilled water with 30% volume of the filtrate to obtain a total of 0.87 L of filtrate and cleaning solution. Then the filtrate and cleaning solution were filtered through an organic membrane, the specification of the organic membrane was 1KDa molecular weight cut-off, and the organic membrane filtrate was washed with 15% volume distilled water to obtain a total of 0.75L of the filtrate and cleaning solution. The obtained combined filtrates were eluted through a Sephadex G-25 gel chromatography column, and the eluent was deionized water. HPLC detects the eluate, collects fractions ...

example 2

[0029] Add 1L of the leech extract stock solution and filter it through a ceramic filter membrane. The specification of the ceramic filter membrane is 150KDa molecular weight cut-off, and wash with distilled water with 30% volume of the ceramic filter membrane filtrate to obtain a total of 0.96L of filtrate and cleaning solution; The liquid passes through a 30KDa ceramic filter membrane, and is cleaned with 30% volume distilled water of the ceramic filter membrane filtrate to obtain a total of 0.8L of the filtrate and the cleaning solution; Wash with 15% volume distilled water of the organic membrane filtrate to obtain a total of 0.68 L of the filtrate and the cleaning solution. The obtained combined filtrates were eluted through a Sephadex G-25 gel chromatography column, and the eluent was deionized water. HPLC detects the eluate, collects fractions with a hirudin chromatographic purity of more than 80%, and obtains 0.47L of the eluate, and then concentrates the eluate throug...

example 3

[0031] Add 1L of the leech extract stock solution and filter it through a ceramic filter membrane. The specification of the ceramic filter membrane is 300KDa molecular weight cut-off, and wash with distilled water with 30% volume of the ceramic filter solution to obtain a total of 0.97L of filtrate and cleaning solution; The liquid passes through a 30KDa ceramic filter membrane, and is cleaned with 30% volumetric distilled water of the ceramic filter membrane filtrate to obtain a total of 0.82L of the filtrate and the cleaning solution; The volume of distilled water was washed, and the organic membrane specification was 1KDa molecular weight cut-off, and the total of filtrate and cleaning solution was 0.7L. The obtained combined filtrates were eluted through a Sephadex G-25 gel chromatography column, and the eluent was deionized water. HPLC detects the eluate, collects the fractions with a hirudin chromatographic purity of more than 80%, and obtains 0.51 L of the eluate, and t...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention discloses a hirudin extraction separation technique which comprises the following steps: smashing and homogenizing hirudo to obtain an extracting solution, filtering through a ceramic filter membrane, and cleaning the ceramic filter membrane with distilled water; merging the ceramic filter membrane filtrate and cleaning solution, filtering through an organic filter membrane, and cleaning the organic filter membrane with distilled water; eluting the organic filter membrane filtrate and cleaning solution through a gel chromatographic column, and detecting the eluate by a high pressure liquid chromatograph; collecting the fraction with the hirudin chromatographical purity of 80% above, concentrating the collected eluate through a nanofiltration membrane, purifying the concentrated solution through a reverse-phase c18 semipreparative column, and collecting the target peak value; and carrying out freeze drying to obtain the freeze-dried hirudin pure product. By adopting the filtration and separation mode of combining the ceramic filter membrane and organic film, most impurities in the hirudo homogenate can be removed, thereby enhancing the extraction efficiency. The technique is convenient to operate, avoids the use of the organic solvent, and greatly lowers the extraction preparation cost.

Description

technical field [0001] The invention relates to a method for filtering and extracting anticoagulant compounds, in particular to an extraction and separation process for hirudin. Background technique [0002] Hirudin is an anticoagulant substance extracted from blood-sucking leeches (commonly known as leeches). Leeches were first recorded in my country's "Shen Nong's Materia Medica", and later in monographs such as "Compendium of Materia Medica" and "Chinese Animal Medicine". In 1884, Haycraft discovered that the leech extract had anticoagulant effect. In 1955, MarkwMdt pointed out that the anticoagulant substance was a protein and named it hirudin. After 1986, genetically recombined hirudin came out, which was the in-depth research and clinical practice of hirudin. Applications open up vast prospects. According to literature records, natural hirudin is a low-molecular-weight polypeptide composed of 65 amino acids, and its functions have the following aspects: [0003] 1. ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/815C07K1/36C07K1/34C07K1/20
Inventor 虞龙张旭琛许友鹏
Owner JINGJIANG ZHIGAO BIOLOGICAL SCI & TECH CO LTD
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap