Rapid determination method of vitality of spleen aminopeptide oral lyophilized powder

A technology for rapid determination and lyophilization of powder, which can be used in the determination/inspection of microorganisms, biochemical equipment and methods, measuring devices, etc., and can solve problems such as laborious and time-consuming

Pending Publication Date: 2019-12-27
DALIAN BAILITIANHUA PHARMA
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This method is time-consumin...

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  • Rapid determination method of vitality of spleen aminopeptide oral lyophilized powder
  • Rapid determination method of vitality of spleen aminopeptide oral lyophilized powder
  • Rapid determination method of vitality of spleen aminopeptide oral lyophilized powder

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Embodiment Construction

[0015] The present invention will be further explained below in conjunction with specific examples.

[0016] (1) Take 3-5 mice with a body weight of 22-25g, both male and female (female is sterile), kill them by neck dislocation, immerse them in 75% alcohol, disinfect the body surface, put them in a purification workbench, and take out the spleen by aseptic operation , placed in a sterilized nylon mesh bag inside the sterilized plate. Add an appropriate amount of RPMI-1640 solution, grind it with a sterilized needle core, and absorb it into a sterilized small test tube. Slowly layer the spleen cell suspension on the lymphocyte separation medium, make the centrifugal force reach 2000r / min within 20 seconds on a horizontal centrifuge, centrifuge for 20min, absorb the lymphocyte layer, suspend with an appropriate amount of 1640 culture medium, 1500r / min, Centrifuge for 5 minutes, and wash twice in this way. Count lymphocytes and adjust the concentration to 3×10 with 1640 cultur...

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Abstract

The invention discloses a rapid determination method of vitality of spleen aminopeptide oral lyophilized powder. When the spleen aminopeptide oral lyophilized powder and splenic lymphocytes of healthymice are incubated together, immunocompetence of the spleen aminopeptide oral lyophilized powder can be adopted to the splenic lymphocytes of the mice to make the splenic lymphocytes of the mice be sensitized lymphocytes. The sensitized lymphocytes release leukocyte adhesion inhibitors, so that the leukocyte adhesion inhibition index is increased. Therefore, the vitality of a finished product canbe verified by determining the leukocyte adhesion inhibition index. According to the rapid determination method, methyl thiazolyl tetrazolium (MTT) is introduced since that the MTT can be reduced toa blue-violet compound (Formazan) by succinodehydrogenase in living cell mitochondria, the number of the reduction product is proportional to the number of cells within a certain range, the Formazan deposited in the cells is dissolved through dimethyl sulfoxide, the A value of the 490 nm wavelength is determined through an enzyme-linked detector, the value is substituted into a formula, and thus the adhesion inhibition index can be solved. The rapid determination method is simple, and detection data are more accurate.

Description

technical field [0001] The invention relates to a determination method, in particular to a rapid determination method for the activity of spleen aminopeptide oral freeze-dried powder. Background technique [0002] The method for measuring the activity of the spleen aminopeptide solution is to take 40ml of blood from the heart of a rabbit, separate the white lymphocytes with the lymphocyte separation medium as the white blood cells for the experiment, and then use the agarose punching method to utilize the biological activity of the white blood cells to move. CO 2 Incubate in a 37°C incubator for 24 hours and observe, and measure the immune activity by the distance that leukocytes move in the agarose. This method requires 40ml of rabbit heart blood for one experiment, and 3-4 rabbits need to be collected. The technical requirements for blood collection are very high, and a full-time teacher is required to be responsible for blood collection, and the demand for experiments i...

Claims

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Application Information

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IPC IPC(8): C12Q1/02G01N21/31
CPCG01N33/5047G01N21/31
Inventor 秦伟刘琦韩树璘衣学龙
Owner DALIAN BAILITIANHUA PHARMA
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