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Cigarette mainstream smoke genotoxicity measurement method

A technology for mainstream cigarette smoke and measurement methods, which is applied in the measurement/inspection of microorganisms, biochemical equipment and methods, and analysis by causing chemical reactions to occur in materials, etc. Gas composition, etc.

Active Publication Date: 2009-11-18
ZHENGZHOU TOBACCO RES INST OF CNTC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Such a test method takes a lot of time and is cumbersome to operate, and because it only tests the particulate matter in mainstream cigarette smoke and does not test the gas phase components in mainstream smoke, it cannot reflect the true genotoxicity of mainstream smoke

Method used

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  • Cigarette mainstream smoke genotoxicity measurement method
  • Cigarette mainstream smoke genotoxicity measurement method
  • Cigarette mainstream smoke genotoxicity measurement method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Evaluate a certain domestic cigarette (package label tar 15mg). The sample cigarettes were smoked under ISO standard conditions, with a puff volume of 35ml per puff, a duration of 2s, and one puff per minute. The mainstream smoke produced by cigarette combustion is diluted with clean air, and the concentration of total particulate matter in the smoke is adjusted to 41.5μg / L, 53.8μg / L, and 67.3μg / L, and the diluted smoke is directly introduced into the The culture flasks of cells (including two groups of samples of -S9 group and +S9 group) were exposed, and the exposure time of each concentration was 60 min. For the connection between the smoking machine used in the test and the cell culture bottle and the smoke control method figure 1 .

[0023] In addition, the culture flask with cells (including two groups of samples of -S9 group and +S9 group) was ventilated with clean air as a control sample. After the exposure, the culture medium in the culture bottle was replac...

Embodiment 2

[0025]This embodiment is basically the same as Embodiment 1, only the cigarette samples and smoke exposure concentrations are different. The cigarette samples were domestic cigarettes with 8mg tar on the box, and the smoke exposure concentrations were 24.2μg / L, 50.6μg / L and 98.7μg / L, see the results in image 3 .

Embodiment 3

[0027] This embodiment is basically the same as Embodiment 1, only the cigarette samples and smoke exposure concentrations are different. The cigarette samples were imported cigarettes with 9 mg tar on the box, and the smoke exposure concentrations were 57.5 μg / L and 90.5 μg / L, see the results in Figure 4 .

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Abstract

A cigarette mainstream smoke genotoxicity measurement method is characterized in that: the cultured cells are directly placed in cigarette mainstream smoke diluted by pure air to expose, the exposed cells are cultured, centrifuged and fixed by cold methanol, acridine orange solution is used for the fluorescent staining for the cells, the stained cells are observed in low power lens to select the region which is distributed evenly and stained well and calculate the number of cells with microkernel(including mononuclear, binuclear and multinuclear) of 500 cells in each sample under fluorescent microscope, the samples is compared with the control sample exposed in pure air to judge whether the cigarette mainstream smoke can increase the cell micronucleus and judge the genotoxicity of the cigarette mainstream smoke. The genotoxicity of the cigarette mainstream smoke can be quantitative evaluated by the invention and the invention has simple operation, high sensitivity and reliable result, thus establishing a new method for measuring cigarette mainstream smoke genotoxicity.

Description

technical field [0001] The invention relates to the field of genotoxicity evaluation of cigarette mainstream smoke, in particular to a method for measuring genotoxicity of cigarette mainstream smoke, which is to measure the proportion of cell micronucleus increase caused by mainstream smoke by exposing cells to cigarette mainstream smoke for genotoxicity assays. Background technique [0002] Cigarette smoke is an aerosol composed of thousands of chemical substances, many of which have certain toxic effects on cells or the human body. Scientists have carried out many studies on the genotoxicity of cigarette smoke or chemical components in the smoke. The CORESTA in vitro toxicology task force recommends the selection of appropriate mammalian cells to evaluate the genotoxicity of cigarette smoke with micronucleus experiments. The purpose of genotoxicity testing is mainly to test the potential negative effects of exogenous substances on the structure or function of DNA. At pre...

Claims

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Application Information

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IPC IPC(8): C12Q1/06G01N21/76
Inventor 卢斌斌宗永立李鹏孙世豪
Owner ZHENGZHOU TOBACCO RES INST OF CNTC
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