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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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  • Summary
  • 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 stable small molecular substances, and the loss of active ingredients is relatively large ; Although affinity chromatography has better selectivity, there are also time-consuming steps such as column packing and pretreatment, and the processing volume is small

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

Experimental program
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Embodiment 1

[0024] Embodiment 1: the preparation of chitosan-histidine endotoxin adsorption medium (MCM-His)

[0025] (1) Ultrasonic dispersion of 1.5g ferroferric oxide in 100mL of acetic acid solution with a volume fraction of 2%, adding 3g chitosan (produced by Aoxing Company, molecular weight 300,000, deacetylation degree 95%, i.e. amino density is 5833 μ mol / g) was completely dissolved and dispersed into 100 mL of liquid paraffin containing 1 mL of Span 80, stirred for 30 min, and then added dropwise with 3 mL of glutaraldehyde solution with a volume fraction of 25%, reacted at 40°C for 2 h, and used a mass fraction of 10% Adjust the pH of the NaOH solution to about 12, and at the same time raise the temperature to 70°C, continue to stir and react for 2h, the product is washed with petroleum ether and hot ethanol, and then vacuum-dried to obtain magnetic chitosan microspheres (MCM), which are determined by acid-base titration. Amino density: Disperse 50mg of microspheres into 20ml o...

Embodiment 2

[0028] Embodiment 2: the preparation of chitosan-tetraethylenepentamine endotoxin adsorption medium (MCM-TEPA)

[0029] (1) Ultrasonic dispersion of 2.0g ferric ferric oxide in 100mL of 5% acetic acid solution by volume fraction, adding 4g chitosan (produced by Aoxing Company, molecular weight 300,000, deacetylation degree 95%, i.e. amino density is 5833 μ mol / g) was completely dissolved and then dispersed into 200mL of liquid paraffin containing 3mL of Span 80. After stirring for 30min, 4mL of glutaraldehyde solution with a volume fraction of 25% was added dropwise and reacted at 40°C for 1h. Adjust the pH of the NaOH solution to about 12, and at the same time raise the temperature to 70°C, continue to stir and react for 3 hours, wash the product with petroleum ether and hot ethanol, and then dry it in vacuum to obtain magnetic chitosan microspheres.

[0030] (2) Disperse 0.5 g of magnetic chitosan microspheres obtained in step (1) into a mixed solution of 10 mL dimethyl sul...

Embodiment 3

[0034] Example 3: Use of Adsorption Media

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

[0036]The endotoxin content was determined by the limulus reagent chromogenic matrix method, and the adsorption kinetic curve of magnetic chitosan microspheres-histidine (MCM-His) to endotoxin was as follows: Figure 4 The adsorption effects of different adsorption media are shown as Figu...

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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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