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Collagen-based biological material endotoxin extraction and detection method

A technology of biological materials and detection methods, applied in the field of biological detection of biological materials, can solve problems such as effective extraction and detection methods that have not been demonstrated, and achieve the effect of removing the shielding effect and improving the detection rate.

Pending Publication Date: 2022-04-26
EXCELLENCE MEDICAL TECH SUZHOU CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

After searching, there is currently no proven effective extraction and detection method for collagen-based medical materials in the form of membranes or solids

Method used

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  • Collagen-based biological material endotoxin extraction and detection method
  • Collagen-based biological material endotoxin extraction and detection method
  • Collagen-based biological material endotoxin extraction and detection method

Examples

Experimental program
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Effect test

Embodiment 1

[0033] In this example, the photometric method of LAL reagent is used to detect endotoxin to illustrate the content of the present invention.

[0034] Weigh 0.02g of pig small intestinal submucosa membrane (experimental group 1) obtained through conventional decellularization method and porcine small intestinal submucosa membrane (experimental group 2) obtained through excessive decellularization method, cut into pieces and place in Add extraction medium (glycerol phosphate, final concentration 0.02% w / t) into the grinding container, mix well and let stand for 2 minutes, fully grind for 5 minutes, centrifuge at 12000rpm for 5 minutes, take the supernatant and dilute it with water for endotoxin inspection for the sample to be tested. Experimental groups 3 and 4 are test groups for conventional endotoxin detection methods for samples of experimental groups 1 and 2, respectively, and only use water for endotoxin detection. Get the sample to be tested, add detection medium (sodiu...

Embodiment 2

[0059] For materials from different tissue sources, the various components contained in them are quite different, such as protein content, fat content, processing reagent residues and other factors may affect the test results. In this embodiment, taking the dermal matrix and the submucosa of porcine small intestine as examples, the test concentration ranges of commonly used surfactants suitable for the present invention are summarized.

[0060] Test method: Weigh a certain amount of porcine small intestinal submucosa and porcine dermal matrix obtained through conventional decellularization methods, cut them into pieces, put them in a grinding container, add extraction medium, mix well and let stand for 2 minutes. Fully grind for 5 minutes, centrifuge at 12000rpm for 5 minutes, take the supernatant and dilute the sample with water for endotoxin test. Take the sample to be tested, add the detection medium and mix well, and then test it on the machine immediately. After reviewin...

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Abstract

The invention discloses a collagen-based biological material endotoxin extraction and detection method. The method comprises the steps of endotoxin extraction and endotoxin detection. The endotoxin extraction comprises the steps of grinding, centrifuging and diluting; wherein an extraction medium is added during grinding of the biological material; the endotoxin detection comprises the step of adding a detection medium into a diluted sample to obtain a solution to be detected. The endotoxin detection method provided by the invention is specially used for collagen-based biological materials.

Description

technical field [0001] The invention belongs to the field of biological detection of biological materials, in particular to a method for extracting and detecting endotoxin from collagen-based biological materials. Background technique [0002] Bacterial endotoxin is a unique structure on the outer layer of the cell wall of Gram-negative bacteria, and its active ingredient is mainly lipopolysaccharide (LPS). When endotoxin enters the human body, it may cause serious consequences such as fever, microcirculation disturbance, endotoxemia, septic shock and disseminated intravascular coagulation. Therefore, in GB / T14233.2-2005 "Medical Infusion, Blood Transfusion, and Injection Equipment Inspection Methods Part 2: Biological Test Methods", it is clearly stipulated that the endotoxin in each infusion, blood transfusion, and injection equipment should not exceed 20EU, and the endotoxin in contact with cerebrospinal fluid and chest cavity Each medical device used within the country ...

Claims

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Application Information

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IPC IPC(8): G01N21/78G01N21/82G01N33/48G01N1/28G01N1/38
CPCG01N21/78G01N21/82G01N33/48G01N1/286G01N1/38G01N2001/2866
Inventor 王妍妍程文悦
Owner EXCELLENCE MEDICAL TECH SUZHOU CO LTD