A method for evaluating the cytotoxicity of e-cigarette liquid based on lactate dehydrogenase assay

Abstract

The invention relates to an electronic cigarette smoke solution cell toxicity evaluation method based on lactate dehydrogenase (LDH) assaying. The method comprises the following steps: (1) preparing an experiment reagent; (2) carrying out cell inoculation; (3) contaminating an electronic cigarette smoke solution; (4) assaying LDH; (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; the sensitivity of detection is improved through optimizing cell inoculating density, selecting a cell lysing solution and optimizing the reaction time of an LDH matrix solution; 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; through assaying LDH, the degree of damage to cell membranes caused by the electronic cigarette smoke solution can be reflected, so that the cell toxicity mechanism of electronic cigarette is disclosed. In addition, the method further has the advantages of quickness, simplicity and convenience in operation, high sensitivity and stable results.

Description

technical field [0001] The invention relates to the research field of external toxicological evaluation of e-cigarette liquid, in particular to a method for measuring the cytotoxicity of e-cigarette liquid based on lactate dehydrogenase. 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 the continuous expansion of consumer grou...

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Problems solved by technology

[0005] At present, the impact of e-cigarette liquid exposure on the integrity of the cell membrane has not been reported in the literature. It is urgent to establish a method for evaluating the cytotoxicity of e-cigarette liquid based on the determination of LDH, to investigate the mechanism of e-liquid cytotoxicity, and to evaluate it quantitatively and accurately Cytotoxicity of e-liquids from commercially available e-cigarette products

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