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A kind of allergy test method for Acanthopanax injection

A technology of Acanthopanax injection and inspection method, which is applied in the direction of microbial measurement/inspection, biochemical equipment and methods, instruments, etc., and can solve the problems of inaccurate reflection of actual sensitization conditions, long time-consuming, large differences, etc. , to achieve convenient and intuitive evaluation and judgment, reduce the risk of occurrence, and improve the effect of comparison standards

Active Publication Date: 2019-07-16
HARBIN ZHENBAO PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the currently commonly used related testing methods are time-consuming; they do not have universal applicability to all types of injections, especially for Acanthopanax injection, and the existing related testing methods cannot accurately reflect its actual toxicity. In the case of allergies, the obtained test data is not stable, the difference between several parallel tests is large, and the repeatability is not good; and the existing test method is used to test some Acanthopanax injections that can cause allergic reactions in mice. , the test result obtained is smaller than that of the negative control, indicating that the existing test technology cannot accurately reflect the sensitization of Acanthopanax injection

Method used

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  • A kind of allergy test method for Acanthopanax injection
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  • A kind of allergy test method for Acanthopanax injection

Examples

Experimental program
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Effect test

Embodiment 1

[0046] Embodiment 1 A kind of anaphylactoid test method of Acanthopanax injection

[0047] The steps are:

[0048] (1), sample pretreatment: put the Acanthopanax injection in a 40°C water bath for 30 minutes, and immediately centrifuge it at 6000rpm for 60s, take the supernatant, and dilute the supernatant 10 times with phenol red-free serum-free MEM medium, Obtain the liquid to be tested;

[0049] (2), prepare detection cell: use the MEM culture that contains 1% penicillin-streptomycin double antibiotics and 10% heat-inactivated fetal bovine serum to cultivate RBL-2H3 cells based on 37 ℃, 5% CO2 cell incubator, when When the RBL-2H3 cells grow to cover 90% of the bottom area of ​​the culture flask, they are subcultured at a ratio of 1:5, once every other day, and then subcultured 3 times to obtain normally growing RBL-2H3 cells, which are the test cells;

[0050] (3), β-hexosaminidase release rate detection: according to 2 × 10 5 Inoculation density per well The test cells...

Embodiment 2

[0058] Embodiment 2 A kind of anaphylactoid test method of Acanthopanax injection

[0059] The steps are:

[0060] (1), sample pretreatment: put Acanthopanax injection in a water bath at 45°C for 25 minutes, then centrifuge at 7000rpm for 50s immediately, take the supernatant, and dilute the supernatant 11 times with phenol red-free serum-free MEM medium, Obtain the solution to be tested.

[0061] (2), prepare detection cell: use the MEM culture that contains 1% penicillin-streptomycin double antibiotics and 10% heat-inactivated fetal bovine serum to cultivate RBL-2H3 cells based on 37 ℃, 5% CO2 cell incubator, when When the RBL-2H3 cells grow to cover 90% of the bottom area of ​​the culture flask, they should be subcultured at a ratio of 1:6, once every other day, and then twice, to obtain normal growing RBL-2H3 cells, which are the test cells;

[0062] (3), β-hexosaminidase release rate detection: according to 5 × 10 5 Inoculation density per well The test cells obtained ...

Embodiment 3

[0070] Embodiment 3 A kind of anaphylactoid test method of Acanthopanax injection

[0071] The steps are:

[0072] (1), sample pretreatment: put Acanthopanax injection in a water bath at 50°C for 20 minutes, then centrifuge it at 8000rpm for 40s immediately, take the supernatant, and dilute the supernatant 12 times with phenol red-free serum-free MEM medium, Obtain the solution to be tested.

[0073] (2), prepare detection cell: use the MEM culture that contains 1% penicillin-streptomycin double antibiotics and 10% heat-inactivated fetal bovine serum to cultivate RBL-2H3 cells based on 37 ℃, 5% CO2 cell incubator, when When the RBL-2H3 cells grow to cover 90% of the bottom area of ​​the culture flask, they should be subcultured at a ratio of 1:7, once every other day, and then twice, to obtain normal growing RBL-2H3 cells, which are the test cells;

[0074] (3), β-hexosaminidase release rate detection: according to 3 × 10 5 Inoculation density per well The test cells obtained...

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Abstract

The invention belongs to the field of pharmaceutical preparations, and in particular relates to an allergy-like testing method for Ciwujia injection. The test method provided by the invention comprises the following steps: (1), sample pretreatment; (2), preparation of detection cells; (3), detection of release rate of β-hexosaminidase. Compared with the prior art, the anaphylactoid test method of Acanthopanax injection provided by the present invention is simple to operate, and the test time is short; it makes the evaluation and judgment of the test results more convenient, intuitive and reliable; using the Acanthopanax provided by the present invention The anaphylactoid test method for injection can obtain relatively accurate test and judgment results that are consistent with the actual mouse experiment; it is more stable and reliable, there is no obvious difference in multiple test results for the same sample, and the repeatability is high.

Description

technical field [0001] The invention belongs to the field of pharmaceutical preparations, and in particular relates to an allergy-like testing method for Ciwujia injection. Background technique [0002] In recent years, with the widespread use of traditional Chinese medicine injections, the reports of adverse reactions have also increased year by year. According to statistics, injections only account for 3% of traditional Chinese medicine preparations, but their adverse reactions are as high as 70%. Adverse reactions of traditional Chinese medicine injections are caused by various reasons, most of which are allergic-like reactions, namely anaphylactoid reactions. The most important difference between anaphylactoid reactions and anaphylaxis is that it occurs without the involvement of the immune system. Drugs can directly stimulate the degranulation of mast cells and basophils in the body, releasing a large number of biologically active substances, such as histamine and leuk...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/34C12Q1/02
CPCC12Q1/34G01N33/5014
Inventor 方同华徐秀杰贾文娟周广红王维龙王博冯慧董丽丽
Owner HARBIN ZHENBAO PHARMA
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