Endotoxin adsorption medium based on magnetic chitosan microballoon and its application method

A technology of chitosan microspheres and adsorption media, applied in the field of biomedical materials, can solve the problems of small processing capacity and large loss of active ingredients, and achieve high selectivity and easy operation

Inactive Publication Date: 2012-02-15
ZHEJIANG UNIV OF TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] At present, the commonly used methods for removing endotoxin include activated carbon adsorption, ultrafiltration, extraction, affinity chromatography, etc., but the first three methods are only suitable for removing endotoxin from relatively stabl

Method used

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  • Endotoxin adsorption medium based on magnetic chitosan microballoon and its application method
  • Endotoxin adsorption medium based on magnetic chitosan microballoon and its application method
  • Endotoxin adsorption medium based on magnetic chitosan microballoon and its application method

Examples

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Example Embodiment

[0024] Example 1: Preparation of Chitosan-Histidine Endotoxin Adsorption Medium (MCM-His)

[0025] (1) Disperse 1.5g of ferroferric oxide ultrasonically in 100mL of 2% acetic acid solution, and add 3g of chitosan (produced by Aoxing Company, with a molecular weight of 300,000, and a degree of deacetylation of 95%, that is, the amino group density is 5833μmol / g) Dissolve it completely and disperse it into 100mL liquid paraffin containing 1mL Span 80, stir for 30min, add 3mL glutaraldehyde solution with a volume fraction of 25%, react at 40°C for 2h, use a 10% mass fraction Adjust the pH of the NaOH solution to about 12, while heating to 70°C, continue to stir and react for 2h, the product is washed with petroleum ether and hot ethanol and dried in vacuum to obtain magnetic chitosan microspheres (MCM), which is determined by acid-base titration Amino density: Take 50mg of microspheres and disperse into 20ml of 10mmol / L hydrochloric acid solution, oscillate for 1h, apply a magnetic...

Example Embodiment

[0028] Example 2: Preparation of Chitosan-Tetraethylenepentamine Endotoxin Adsorption Medium (MCM-TEPA)

[0029] (1) Disperse 2.0 g of ferroferric oxide ultrasonically in 100 mL of 5% acetic acid solution, and add 4 g of chitosan (produced by Aoxing Company, with a molecular weight of 300,000, and a degree of deacetylation of 95%, that is, the amino group density is 5833μmol / g) Dissolve it completely and then disperse it into 200mL liquid paraffin containing 3mL Span 80. After stirring for 30 minutes, add 4mL glutaraldehyde solution with a volume fraction of 25%, react at 40°C for 1 hour, and use a 10% mass fraction The pH of the NaOH solution was adjusted to about 12, while the temperature was raised to 70°C, and the stirring reaction was continued for 3 hours. The product was washed with petroleum ether and hot ethanol and dried in vacuum to obtain magnetic chitosan microspheres.

[0030] (2) Disperse 0.5 g of the magnetic chitosan microspheres obtained in step (1) into 10 mL of...

Example Embodiment

[0034] Example 3: Use of adsorption medium

[0035] Take magnetic chitosan microspheres (MCM prepared in Example 1), magnetic chitosan microspheres-histidine (MCM-His prepared in Example 1), and magnetic chitosan microspheres-tetraethylene five Amine (MCM-TEPA prepared in Example 2) 50mg was washed with a mass fraction of 1% sodium deoxycholate aqueous solution, suction filtration or magnetic field separation three times, then washed with pyrogen-free water three times, dried and dispersed into 2mL solution containing endotoxin (Tap water), shake and adsorb for 30 minutes, absorb the adsorption medium with a magnet, and take out the supernatant to determine the endotoxin content.

[0036] The endotoxin content is determined by the limulus reagent color matrix method, and the adsorption kinetic curve of the magnetic chitosan microsphere-histidine (MCM-His) on endotoxin is as follows Figure 4 As shown, the adsorption effect of different adsorption media is as Figure 5 Shown from ...

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Abstract

The invention discloses an endotoxin adsorption medium based on magnetic chitosan microballoon and an application method, wherein the preparation method of endotoxin adsorption medium comprises the following steps: (1) dispersing a magnetic material in an acetic acid solution, adding chitosan to prepare water phase I, transferring water phase I to liquid paraffin containing span 80, taking glutaraldehyde as a cross-linking agent, preparing chitosan microballoon by using an anti-phase suspension method; (2) dispersing magnetic chitosan microballoon in a mixed solution of dimethyl sulfoxide and sodium hydroxide, adding chloropropylene oxide for activating, washing by deionized water till neutralizing to obtain an activated product; (3) placing the activated product in an endotoxin affinity ligand solution for immobilizing to obtain the endotoxin adsorption medium based on the magnetic chitosan microballoon. The endotoxin adsorption medium of the present invention has high selectivity of endotoxin and simple operation, and can be used for removing the endotoxin in a plurality of biological products.

Description

1. Technical field [0001] The invention relates to the field of biomedical materials, in particular to the preparation and use method of an in vitro endotoxin adsorption medium. 2. Background technology [0002] Endotoxin is a unique structure on the outer membrane of the cell wall released by Gram-negative bacteria when they grow, divide or die. Because Gram-negative bacteria are very stubborn and only need very little nutrients to survive in water, endotoxins are widely present in drinking water, food, biomedical preparations, and on the walls of devices in contact with aqueous solutions; in higher organisms, stomach Gram-negative bacteria in the lower intestinal tract are the main source of biological endotoxins, and the endotoxins released after the bacteria die will enter the blood circulation and cause a series of pathological changes. If the bacterial endotoxin in the injection is greater than 1EU / mL in the clinical use of the hospital, the patient will have chills, ...

Claims

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Application Information

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IPC IPC(8): B01J20/24B01J20/28B01J20/30
Inventor 应国清来灿林易喻梅建凤王鸿陈建澍姜玉奇
Owner ZHEJIANG UNIV OF TECH
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