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Preparation method of bacterial cellulose capable of removing endotoxin

A technology of bacterial cellulose and bacterial cellulose membrane, which is applied in the field of bacterial cellulose preparation, can solve the problems of high price, limited sample size, and difficult removal of endotoxin, etc., and achieves the effect of simple preparation process and simple method

Inactive Publication Date: 2013-03-20
SHANGHAI DIVINE MEDICAL TECH
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  • Abstract
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AI Technical Summary

Problems solved by technology

Endotoxins produced by Gram-negative bacteria are not easily removed during polysaccharide purification
The endotoxin removal process recommended by the World Health Organization (WHO) in Gram-negative bacterial polysaccharide products is ultracentrifugation for 4-6 hours under the condition of a centrifugal force of 100,000g, but this process not only requires a very expensive ultra-fast refrigerated centrifuge , and the amount of sample processed at one time is limited
There are also many endotoxin removal methods reported in the literature. According to different modes of action, they can be divided into the following three categories: ① non-selective methods, mainly including ultrafiltration, activated carbon adsorption, and phase separation; ② chromatography, mainly including Ion exchange chromatography, hydrophobic chromatography and gel filtration chromatography; ③selective methods, mainly refer to affinity adsorption method, also include metal chelate adsorption method and aluminum hydroxide adsorption method, but these methods are not suitable for medical treatment Biomaterials for devices are not good methods

Method used

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  • Preparation method of bacterial cellulose capable of removing endotoxin

Examples

Experimental program
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Effect test

Embodiment 1

[0027] The commercially available bacterial cellulose membrane prepared by Acetobacter xylinum was washed with purified water several times to control the pH value of the bacterial cellulose to 7-7.2 to obtain a pretreated bacterial cellulose membrane, which was refrigerated for future use.

[0028] Prepare 40%wt PEG200 aqueous solution with purified water.

[0029] Soak the bacterial cellulose membrane in PEG aqueous solution, heat it in a water bath to 80°C, and keep it warm for 0.5h.

[0030] After cooling, the PEG / bacterial cellulose composite membrane was taken out and washed 3 times with purified water.

[0031] For endotoxin detection, the bacterial endotoxin limit is not more than 20EU per piece, and the sensitivity of the LAL reagent is 1EU / ml.

Embodiment 2

[0033] The commercially available bacterial cellulose membrane prepared by Acetobacter xylinum was washed with purified water several times to control the pH value of the bacterial cellulose to 7-7.2 to obtain a pretreated bacterial cellulose membrane, which was refrigerated for future use.

[0034] Prepare 50%wt PEG400 aqueous solution with purified water.

[0035] Soak the bacterial cellulose membrane in PEG aqueous solution, heat it in a water bath to 90°C, and keep it warm for 1h.

[0036] After cooling, the PEG / bacterial cellulose composite membrane was taken out and washed 3 times with purified water.

[0037] The obtained PEG / bacterial cellulose composite membrane was sterilized by high temperature and moist heat.

[0038] For endotoxin detection, the bacterial endotoxin limit is not more than 20EU per piece, and the sensitivity of the LAL reagent is 1EU / ml.

Embodiment 3

[0040] The commercially available bacterial cellulose membrane prepared by Acetobacter xylinum was washed with purified water several times to control the pH value of the bacterial cellulose to 7-7.2 to obtain a pretreated bacterial cellulose membrane, which was refrigerated for future use.

[0041] Prepare 75%wt PEG800 aqueous solution with purified water.

[0042] Soak the bacterial cellulose membrane in PEG aqueous solution, heat it in a water bath to 100°C, and keep it warm for 1.5h.

[0043] After cooling, the PEG / bacterial cellulose composite membrane was taken out and washed 3 times with purified water.

[0044] The obtained PEG / bacterial cellulose composite membrane was sterilized by irradiation, and the dose of irradiation sterilization was 5KGy.

[0045] For endotoxin detection, the limit of bacterial endotoxin is not more than 2.15EU per piece, and the sensitivity of LAL is 0.25EU / ml.

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Abstract

The invention discloses a preparation method of bacterial cellulose capable of removing endotoxin. The preparation method comprises the following steps of: pretreating a bacterial cellulose membrane and preparing a PEG (polyethylene glycol) water solution or chitosan solution; putting the bacterial cellulose membrane in the PEG water solution or chitosan solution, heating to above 80 DEG C and maintaining at 80-100DEG C under a water bath for 0.5-1.5h; and washing the cooked bacterial cellulose membrane with purified water to obtain the bacterial cellulose. The method for removing the endotoxin is simple without considering the removal of the endotoxin in the bacterial cellulose culture process; and the preparation method is simple in preparation process and is suitable for industrial production of composite bacterial cellulose dressing.

Description

technical field [0001] The invention relates to a preparation method of bacterial cellulose. Background technique [0002] When using drug injections or medical devices in hospital clinics, cold sensation, chills, fever, headache, nausea, vomiting, pale complexion, shock, and death may occur in severe cases. This symptom is called pyrogen reaction, which is caused by microbial contamination. The pyrogen reaction of endotoxin is called endotoxin pyrogen. Bacterial endotoxin is a complex of lipopolysaccharide and trace protein on the cell wall of Gram-negative bacteria. Its chemical composition is mainly composed of three parts: O-specific chain, core polysaccharide, and lipid A. Lipid A is the inner The main toxic component of the toxin. [0003] Endotoxin is widely distributed in the lipopolysaccharide of the cell wall layer of Gram-negative bacteria and other microorganisms. Endotoxins produced by Gram-negative bacteria are not easily removed during polysaccharide purifi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08J7/00C08L1/02
Inventor 薛燕万隆沈静飞
Owner SHANGHAI DIVINE MEDICAL TECH
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