Method of detecting influence on human oral cell [gamma] H2AX level due to tobacco chewing gum

A tobacco product, gum-based technology, applied in the field of biotechnology applications, can solve problems such as inability to accurately conduct biosafety testing, unsuitable cigarette safety assessment, and difficulty in dissolving the contents, so as to improve efficiency and reliability. , the effect of improving the accuracy

Inactive Publication Date: 2017-02-01
CHINA TOBACCO YUNNAN IND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, gum-based tobacco products do not produce smoke and directly affect the oral cavity, so the safety assessment methods of traditional cigarettes are not applicable to this product
Snus products in pouches are generally used by sticking them between the gums and lips,

Method used

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  • Method of detecting influence on human oral cell [gamma] H2AX level due to tobacco chewing gum

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0037] 1. Randomly select a gum-based tobacco product (N-2, nicotine content 2mg / piece), randomly select 2 pieces and put them into a sample tube, and freeze the sample tube in liquid nitrogen for 30 minutes.

[0038] 2. Take out the sample tube from the liquid nitrogen, and then take out the gum-based tobacco product (N-2), immediately pulverize it with a sample pulverizer, and collect the pulverized powder with the sample tube.

[0039] 3. Place the sample tube containing the powdered gum-based tobacco products in liquid nitrogen and freeze for 30 minutes;

[0040] 4. Take out the sample tube from the liquid nitrogen, immediately add 10mL of artificial saliva preheated to 37°C, soak the pulverized gum-based tobacco product powder (N-2), use the extraction conditions of 37°C and 30min, and Stir the extract solution every 2 minutes during the soaking process to make the sample fully contact with the extract solution to facilitate the dissolution of the contents.

[0041] 5. P...

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Abstract

The invention discloses a method of detecting influence on human oral cell [gamma] H2AX level due to tobacco chewing gum. The method includes the steps of: 1) freezing to-be-tested tobacco chewing gum in liquid nitrogen; 2) instantly crushing the tobacco chewing gum by means of a crusher; 3) freezing the crushed tobacco chewing gum in the liquid nitrogen; 4) soaking the crushed tobacco chewing gum in artificial saliva; 5) centrifugally separating an extract liquid; 6) detecting a supernatant liquid by means of an osmotic pressure measuring instrument and regulating the osmotic pressure to be consistent with that of a cell culture medium; 7) measuring the content of nicotine in the extract liquid; 8) culturing human oral mucosa epithelial cells as a detection object; 9) adding the extract liquid of the tobacco chewing gum to the cultured human oral mucosa epithelial cells; and 10) culturing the cells for 4 hours, and detecting the change situation of the [gamma] H2AX of the cells by means of a high-content imaging instrument.

Description

technical field [0001] The invention relates to the application field of biotechnology, in particular to the determination of the effect of gum-based tobacco products on rH2AX of human oral cavity cells. Background technique [0002] Gum-based smokeless tobacco product (Tobacco chewing gum) is an oral smokeless tobacco product made by adding a certain amount of tobacco extract, spices and other edible additives to the gum base. Gum-based tobacco products are used by putting them in the mouth and chewing them. The chewing extracts will merge with saliva and gather in the mouth, and be spit out or swallowed according to different consumption habits. Since the product mainly acts on the oral cavity and directly contacts oral cells, it is necessary to establish a method for detecting its effect on the γH2AX level of human oral cells to evaluate its safety. [0003] The existing safety evaluation methods of tobacco products are mainly aimed at traditional cigarettes, usually by ...

Claims

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Application Information

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IPC IPC(8): C12Q1/02
CPCG01N33/502G01N33/5044
Inventor 高茜李雪梅夭建华朱洲海黄海涛倪红梅管莹米其利刘欣
Owner CHINA TOBACCO YUNNAN IND
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