Neutral red absorption assay method for e-cigarette liquid cytotoxicity evaluation

A technology of e-cigarette liquid and cytotoxicity, which is applied in the direction of color/spectral characteristic measurement, etc., to achieve high sensitivity, stable results, and accurate quantitative effects

Active Publication Date: 2017-01-11
CHINA NAT TOBACCO QUALITY SUPERVISION & TEST CENT
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Problems solved by technology

However, whether this method can be used to evaluate the cytotoxicity of e-cigarette liquid has not been reported in the literature

Method used

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  • Neutral red absorption assay method for e-cigarette liquid cytotoxicity evaluation
  • Neutral red absorption assay method for e-cigarette liquid cytotoxicity evaluation
  • Neutral red absorption assay method for e-cigarette liquid cytotoxicity evaluation

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Embodiment Construction

[0026] The present invention will be further described below in conjunction with the embodiments, but it does not limit the present invention.

[0027] To investigate the toxic effect of e-cigarette sample 2 smoke liquid on BEAS-2B cells. After the expanded and cultured BEAS-2B cells were trypsinized, a cell suspension (2×10 5 / Ml). Add 100 μl cell suspension to each well of a 96-well plate (except the 36 wells on the outermost periphery). 2 Culture for 24 h under the conditions. After the cells were cultured for 24 hours, the culture medium was aspirated, and each column of the 96-well plate (except the outermost 36 wells) was set as a negative control and 8 different doses (5 mg / ml, 10 mg / ml, 15 mg). / ml, 20mg / ml, 30 mg / ml, 40 mg / ml, 50 mg / ml, 60 mg / ml) e-cigarette liquid poisoning and positive control treatment. Add 100μl of diluted culture medium to each of the 36 holes on the outermost periphery of the 96-well plate as a solution background control, at 37 ℃, 5% CO 2 Cultu...

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Abstract

The invention relates to a neutral red absorption assaying method for evaluating the cell toxicity of an electronic cigarette smoke solution. The method is characterized by comprising the following steps: (1) preparing an experiment reagent; (2) carrying out cell inoculation; (3) contaminating the electronic cigarette smoke solution; (4) dyeing with neutral red; (5) resulting and analyzing. According to the method, aiming at the characteristic that most of electronic cigarette smoke solution samples are high in density, a method that an electronic cigarette contaminated solution is prepared by adopting mass weighing is determined; by a cell inoculation density optimizing step, cells of a cell line BEAS-2B, which are most applicable to the toxicity evaluation of the electronic cigarette smoke solution under the optimal inoculation density, are screened; the best contaminating concentration is determined through optimizing the contaminating concentration of the electronic cigarette smoke solution, so as to obtain the optimal dose-effect curve. The assaying method disclosed by the invention further has the advantages of quickness, simplicity, convenience and high sensitivity.

Description

Technical field [0001] The invention relates to the determination of the cytotoxicity of the electronic cigarette liquid, in particular to a quantitative determination method based on the neutral red dye analysis of the cell survival rate after the electronic cigarette liquid is poisoned. Background technique [0002] As people understand that "smoking is harmful to health", governments of all countries have actively adopted and implemented effective legislative, administrative or other measures within the scope of current laws and regulations to prohibit smoking in public places. In 2005, China signed the World Health Organization's Framework Convention on Tobacco Control. In 2012, the Chinese government issued the "China Tobacco Control Plan (2012-2015)", which proposed to create a smoke-free environment and implement a total ban on smoking in public places. Indoor smoking bans have led to the rapid growth of new products such as electronic cigarettes in the markets of various...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/31
Inventor 刘彤陈欢韩书磊吴帅宾付立伟张小涛石龙凯侯宏卫胡清源
Owner CHINA NAT TOBACCO QUALITY SUPERVISION & TEST CENT
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