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Method for testing cytotoxicity in full smoke contamination of cigarette

A technology of cytotoxicity and testing methods, which is applied in the direction of biochemical equipment and methods, and microbial determination/inspection, etc., can solve the problem of not directly reflecting the toxic effects of cigarette smoke, not fully reflecting the cytotoxicity of cigarette smoke, and being unable to communicate with cells To avoid problems such as full contact with cigarette smoke, achieve comprehensive and reliable results, reduce fixed steps, and easy operation

Active Publication Date: 2011-08-03
ZHENGZHOU TOBACCO RES INST OF CNTC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In this method, the cells are not in the real smoke environment to directly and real-time experience the fresh mainstream smoke of cigarettes, and the test results cannot directly reflect the toxic effects of actual exposure to cigarette smoke
The patent (publication number CN101393190) is a method for determining the cytotoxicity of mainstream cigarette smoke. The method is to introduce the smoke into a cell culture bottle for exposure, and the cells are indirectly contacted with the smoke components dissolved in the culture medium, while for the insoluble Because the smoke components in the culture medium cannot contact the cells, the test results cannot fully reflect the cytotoxicity of cigarette smoke
The disadvantage of the above two methods is that the cultured cells cannot be directly, comprehensively and fully contacted with cigarette smoke.

Method used

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  • Method for testing cytotoxicity in full smoke contamination of cigarette
  • Method for testing cytotoxicity in full smoke contamination of cigarette
  • Method for testing cytotoxicity in full smoke contamination of cigarette

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1 The cytotoxicity test of whole smoke from a domestic cigarette.

[0054] First, prepare the experimental reagents according to the above step a, and then carry out cell seeding and culture according to the above step b. Inoculate the cultured CHO cells in a 12mm insert cell culture dish (Transwell chamber) with a cell density of 3.5×10 4 / Transwell chamber, at 37℃, 5% CO 2 Incubate for 24 h under the conditions; then carry out the whole cigarette smoke poisoning according to the above step c, that is, transfer the Transwell chamber to the smoke exposure device, and smoke the cigarette sample under deep smoking conditions. The mainstream cigarette smoke produced is passed through the clean air After dilution, it is introduced into the smoke exposure device, and 4 cigarette smoke doses with different dilution ratios are set (the suction volume of cigarette smoke is 55 ml per puff, and the discharge time per puff is 2.8 s; the flow rate of clean air is set to 6 L / mi...

Embodiment 2

[0057] Example 2 A complete smoke cytotoxicity test of a domestic cigarette. Refer to Example 1 for specific implementation.

[0058] The TPM release of a single cigarette is 44.9 mg / stick.

[0059] Cytotoxicity of cigarette smoke

[0060]

Embodiment 3

[0061] Example 3 The whole smoke cytotoxicity test of a certain domestic cigarette. Refer to Example 1 for specific implementation.

[0062] The TPM release of a single cigarette is 31.35 mg / stick.

[0063] Cytotoxicity of cigarette smoke

[0064]

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Abstract

The invention relates to a method for testing cytotoxicity in full smoke contamination of cigarette. In the method, when full smoke exposure of the cigarette is carried out, an insertion type cell culture dish is used so that cultured cells are positioned at a gas-liquid interface between the smoke and the culture solution, and therefore, the purpose that the cells directly and fully contact withcigarette smoke is achieved, and the feel of the cells to the cigarette smoke can be comprehensively reflected. The method has high sensitivity, and the result can more truly reflect the cytotoxicityof the cigarette smoke. The insertion type cell culture dish is used so that a grain-phase part and a gas-phase part in the main-flow smoke of the cigarette directly and fully contact with the cultured cells, and when the full smoke exposure of the cigarette is carried out, the cells growing at microporous membrane attached to a wall are positioned at the gas-liquid interface between the cigarette smoke and the exposed culture solution, so that the problem that the cells can not comprehensively feel the full cigarette smoke in the previous experiments is solved; and meanwhile, in the method, the stationary step for formaldehyde stationary liquid is reduced so that the experimental process is simpler and more convenient.

Description

technical field [0001] The invention relates to the research field of external toxicological evaluation of cigarette smoke, in particular to a cytotoxicity test method for whole cigarette smoke exposure. Background technique [0002] The in vitro toxicological study of cigarette smoke has become one of the important means to evaluate the health hazards of smoking. Cigarette smoke is a complex aerosol composed of thousands of chemical substances, and the smoke components are distributed in the particle phase and gas phase of the smoke aerosol. The main harmful components in the particle phase of smoke are nicotine, aromatic amines, phenols, tobacco-specific nitrosamines, polycyclic aromatic hydrocarbons, etc.; harmful components such as CO, nitrogen oxides and volatile organic compounds are distributed in the gas phase ; Aldehydes, ketone carbonyl compounds, hydrocyanic acid, ammonia and other harmful components are distributed in the gas and particle phase. At the same tim...

Claims

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Application Information

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IPC IPC(8): C12Q1/02
Inventor 李翔尚平平聂聪杨松王宜鹏刘惠民谢剑平
Owner ZHENGZHOU TOBACCO RES INST OF CNTC
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