Neutral red absorption assaying method for evaluating cell toxicity of electronic cigarette smoke solution

A technology for e-cigarette liquid and cytotoxicity, which is applied in the measurement of color/spectral characteristics, etc., to achieve the effects of high sensitivity, high detection throughput, and accurate quantification.

Active Publication Date: 2014-05-21
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 assaying method for evaluating cell toxicity of electronic cigarette smoke solution
  • Neutral red absorption assaying method for evaluating cell toxicity of electronic cigarette smoke solution
  • Neutral red absorption assaying method for evaluating cell toxicity of electronic cigarette smoke solution

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Embodiment Construction

[0026] The present invention will be further described below in conjunction with the examples, but the present invention is not limited.

[0027] In order to investigate the toxic effects of e-cigarette sample 2 e-liquid exposure on BEAS-2B cells. After the expanded and cultured BEAS-2B cells were digested with trypsin, the cell suspension (2×10 5 pieces / ml). Add 100 μl of cell suspension to each well of a 96-well plate (except the outermost 36 wells), and store at 37 °C, 5% CO 2 Incubate under the condition for 24 h. After the cells were cultured for 24 h, the culture medium was sucked off, and each column of 6 wells in a 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 exposure and positive control treatment. Add 100 μl of diluted culture solution to each well of the outermost 36 wells of the 96-well plate as a solution back...

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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 measurement of extracellular cytotoxicity of e-cigarette liquid, in particular to a quantitative determination method for analyzing the cell survival rate of e-cigarette liquid after exposure to poison based on neutral red dye. Background technique [0002] With people's understanding of "smoking is harmful to health", governments of various 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 WHO Framework Convention on Tobacco Control. In 2012, the Chinese government issued the "China Tobacco Control Plan (2012-2015)", proposing to create a smoke-free environment and implement a comprehensive ban on smoking in public places. The indoor smoking ban has led to the rapid growth of new products such as electronic cigarettes in the markets of various countries, and...

Claims

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Application Information

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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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