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Heparin-phenylalanine adsorption material for blood purification method for removing endotoxin

A technology of blood purification and phenylalanine, which is applied in the field of biomedical engineering, can solve the problems of death of Gram-negative bacteria and no safe and effective specific drugs have been found, and achieve a high removal rate

Inactive Publication Date: 2013-08-07
CHONGQING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] Endotoxin is heat-resistant and stable, and no safe and effective specific drug has been found yet, and the use of antibiotics will lead to the death of a large number of Gram-negative bacteria and release more endotoxin

Method used

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  • Heparin-phenylalanine adsorption material for blood purification method for removing endotoxin
  • Heparin-phenylalanine adsorption material for blood purification method for removing endotoxin
  • Heparin-phenylalanine adsorption material for blood purification method for removing endotoxin

Examples

Experimental program
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Effect test

Embodiment 1

[0036] The preparation of embodiment 1 heparin-phenylalanine adsorption material

[0037] (1) Amination of chloromethyl resin

[0038] Take the chloromethyl resin and wash it alternately with absolute ethanol and distilled water three times each, and drain it with a suction filter pump for later use; place 5-10 g of the obtained chloromethyl resin in 20-60 ml of 1,4-dioxane to swell for 2-6 hours, Transfer to a three-necked flask; dissolve 0.1-1g of tetrabutylammonium bromide and 1-10g of NaOH in 40ml of distilled water and add to the three-necked flask; then add 10-50ml of ethylenediamine to the three-necked flask; control the reaction The temperature is 40-100°C, and the reaction is stirred for 1-9 hours; the product obtained from the reaction is filtered, and the filter residue is alternately washed with absolute ethanol and distilled water until neutral, and finally dried in vacuum to obtain an aminated chloromethyl resin;

[0039] (2) Connection of spacer arm heparin

[0...

Embodiment 2

[0043] Example 2 Observation of the adsorption of fluorescent endotoxin (FITC-LPS) by heparin-phenylalanine adsorption material under fluorescence microscope

[0044] Adsorption experiments were carried out on 0.2 g of the heparin-phenylalanine adsorption material prepared in Example 1 in rabbit plasma solutions containing 50 μg / mL fluorescent endotoxin. The adsorption reaction time was 2 hours, and the surface fluorescence intensity was observed by an inverted fluorescence microscope. Qualitatively observe the adsorption situation; under the same experimental conditions, the aminated chloromethyl resin prepared in Example 1 step (1) and the heparinized chloromethyl resin prepared in Example 1 step (2) were used as a control group, and the experimental results were as follows: Figure 2~4 shown. figure 2 It is the effect diagram of heparin-phenylalanine adsorption material adsorbing fluorescently labeled endotoxin, image 3 is the effect diagram of heparinized chloromethyl r...

Embodiment 3

[0045] Example 3 Heparin-phenylalanine adsorption material is used for the removal of endotoxin in rabbit plasma

[0046] Limulus reagent chromogenic matrix method is used for the quantitative detection of endotoxin. The basic principle is that bacterial endotoxin reacts enzymatically with factor C in Limulus reagent to produce coagulation enzyme, which can decompose the artificially synthesized chromogenic matrix into yellow paranitrate Phenylaniline (PNA) and peptides. PNA can be further dyed rose color by azo reagent, and the absorbance value in the reaction solution can be measured at 545nm for endotoxin quantification.

[0047] Take 0.2 g of the heparin-phenylalanine adsorption material prepared in Example 1 and fill it into a 2 mL syringe, plug both ends with absorbent cotton, connect a constant-flow pump and a beaker to make a hemoperfusion device for dynamic plasma perfusion experiments.

[0048] The Limulus kit was used to detect the adsorption amount of the heparin-...

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Abstract

The invention relates to a heparin-phenylalanine adsorption material for a blood purification method for removing endotoxin. The preparation method of the heparin-phenylalanine adsorption material disclosed by the invention comprises the following steps: carrying out amination on chloromethylated polystyrene-diethylbenzene ene resin (merrifield resin) serving as a base material by using ethylenediamine to prepare the amino-methyl chloride resin, then utilizing 1-(3-dimethyl ami-nopropyl)-3-ethyl carbodiimide / N-hydroxysuccinimide (EDC / NHS) as a catalyst to connect spacer heparin, so as to prepare heparinized methyl chloride resin; and finally with the EDC / NHS as a catalyst, connecting phenylalanine to heparin, so as to prepare the heparin-phenylalanine adsorption material. The heparin-phenylalanine adsorption material disclosed by the invention can be effectively applied to the blood purification method for removing endotoxin, and is good in blood compatibility, and safe and nontoxic to a human body.

Description

technical field [0001] The invention belongs to the field of biomedical engineering, in particular to a heparin-phenylalanine adsorption material used for blood purification to remove endotoxin. Background technique [0002] Endotoxin, lipopolysaccharide, is ubiquitous in the human body as a component of the cell wall of Gram-negative bacteria. When the human body is in a state of stress such as severe trauma and infection, the death of bacteria will release a large amount of endotoxin, which will lead to endotoxemia (Endotoxemia, ETM), and then cause systemic inflammatory response syndrome, sepsis, multiple organ dysfunction, etc. handicap syndrome, etc. ETM is characterized by high morbidity, high mortality, and high treatment costs [1]. [0003] Endotoxin is heat-resistant and stable, and no safe and effective specific drug has been found yet, and the use of antibiotics will lead to the death of a large number of Gram-negative bacteria and release more endotoxin. There...

Claims

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Application Information

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IPC IPC(8): B01J20/24B01J20/30A61M1/14
Inventor 王翔靳欣欣杨力毛金春王菲菲
Owner CHONGQING UNIV
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