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Cell toxicity determination method in cigarette mainstream flue gas

A technology for mainstream smoke and cytotoxicity of cigarettes, which is applied to the determination/inspection of microorganisms, biochemical equipment and methods, measuring devices, etc., and can solve the problems of cumbersome operation, long time, and failure to reflect the cytotoxicity of mainstream smoke

Inactive Publication Date: 2009-03-25
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 the mainstream smoke of cigarettes, it does not test the gas phase components in the mainstream smoke, so it cannot reflect the true cytotoxicity of the mainstream smoke.

Method used

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  • Cell toxicity determination method in cigarette mainstream flue gas
  • Cell toxicity determination method in cigarette mainstream flue gas
  • Cell toxicity determination method in cigarette mainstream flue gas

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] 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 burning is diluted with clean air, and the concentration of total particulate matter in the smoke is adjusted to 30 μg / L, 60 μg / L, 90 μg / L and 120 μg / L, and the diluted smoke is directly introduced into the The culture flasks of the cells 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 .

[0021] In addition, clean air was blown into the culture flask with cells as a control sample. After the exposure, the culture medium in the culture bottle was replaced with fresh culture medium and placed in CO 2 Cultivate in the incubator for 24h. The medium was then aspirated and ...

Embodiment 2

[0023] 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 20μg / L, 60μg / L, 80μg / L, 100μg / L, 120μg / L and 150μg / L. see results image 3 . LC 50 The calculated result was 78.0 μg / L.

Embodiment 3

[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 3 mg tar in the box, and the smoke exposure concentrations were 60 μg / L, 90 μg / L, 120 μg / L and 160 μg / L. see results Figure 4 . LC 50 The calculated result was 95.1 μg / L.

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Abstract

The invention relates to a method for mensurating cytotoxicity of mainstream cigarette smoke. The method is characterized in comprising the following steps: firstly, a cultured cell is directly put to the mainstream cigarette smoke diluted by clean air and is exposed; secondly, the cell is subjected to neutral red staining; according to the situation that the cell is ingested with neutral red, a spectrophotometer of a readable porous culturing plate is utilized to mensurate an absorbance value; and the cell is compared with a contrastive sample exposed in the clean air, thereby judging the toxicity of the mainstream cigarette smoke. The method is based on a testing principle that the cultured cell is directly put to the mainstream cigarette smoke and is exposed; the level for ingesting the neutral red by the cell is in positive proportion to the number of viable cells; when the toxic effect of the mainstream cigarette smoke is larger, the survival cells exposed in the mainstream cigarette smoke is fewer, correspondingly the dye absorption amount is less and the absorbance value is lower. The method can quantitatively evaluate the cytotoxicity of a tested object, has the characteristics of simple operation, high sensitivity and reliable result, and creates a new method for mensurating the cytotoxicity of the mainstream cigarette smoke.

Description

technical field [0001] The invention relates to the field of cytotoxicity evaluation of mainstream cigarette smoke, in particular to a method for measuring cytotoxicity of mainstream cigarette smoke, which is to measure the cell death rate caused by mainstream smoke by exposing cells Toxicity Assay. 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 cytotoxicity of cigarette smoke or chemical components in smoke. The CORESTA In vitro Toxicology Task Force recommends the selection of appropriate mammalian cells to evaluate the cytotoxicity of cigarette smoke with the neutral red method. Neutral The red evaluation method has been widely used to evaluate and compare the cytotoxicity of smoke condensate of different tar contents and different types of cigarettes [Putnam KP, Bombick DW, Doolittle DJ. Evaluatio...

Claims

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Application Information

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IPC IPC(8): G01N33/00C12Q1/02
Inventor 卢斌斌宗永立胡军孙世豪
Owner ZHENGZHOU TOBACCO RES INST OF CNTC
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