Method for detecting influences of gum based chewing tobacco on cellular superoxide dismutase activity

A technology of superoxide and dismutase enzymes, which is applied in the direction of measuring devices, biological testing, material inspection products, etc., can solve the problems of time-consuming and labor-intensive, no uniform standard methods, etc., and achieve accurate and effective detection results

Pending Publication Date: 2018-06-05
CHINA TOBACCO YUNNAN IND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are a large number of primary formulas, and sensory evaluation of all of them is time-consuming and labor-intensive, and the sensory evaluation focuses on screening the style and taste of the product. How to use biological test indicators to screen the primary formula of the product to obtain the effect of reducing the adverse feelings of the human body? The product formula is still in the exploratory stage, and there is no uniform standard method

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  • Method for detecting influences of gum based chewing tobacco on cellular superoxide dismutase activity
  • Method for detecting influences of gum based chewing tobacco on cellular superoxide dismutase activity

Examples

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Effect test

Embodiment 1

[0040] A method for detecting the influence of gum-based chewing tobacco on the activity of superoxide dismutase in cells, comprising the following steps:

[0041] Step (1), sample pretreatment: add the gum-based chewing tobacco product to the oral keratinocyte culture medium at 37°C, and the volume ratio of the gum-based chewing tobacco product to the oral keratinocyte culture medium is 1g: 10mL; Grinding at 37°C for 10 minutes, then filtering, adding serum to the filtrate so that the final volume percentage of serum is 10%, to obtain the test solution;

[0042]Step (2), preparation of single cell suspension: HOK human oral keratinocytes at 37°C, 5% CO 2 Oral keratinocyte culture medium was used in the incubator. When the cell confluency rate was 80-85%, the medium was removed, washed twice with phosphate buffer solution with a pH value of 7.2, and the washing solution was discarded; the concentration of addition was 0.25% (w / v) trypsin solution for monolayer incubation for...

Embodiment 2

[0059] A method for detecting the influence of gum-based chewing tobacco on the activity of superoxide dismutase in cells, comprising the following steps:

[0060] Step (1), sample pretreatment: add the gum-based chewing tobacco product to the oral keratinocyte culture medium at 37°C, and the volume ratio of the gum-based chewing tobacco product to the oral keratinocyte culture medium is 1g: 10mL; Grinding at 37°C for 10 minutes, then filtering, adding serum to the filtrate so that the final volume percentage of serum is 10%, to obtain the test solution;

[0061] Step (2), preparation of single cell suspension: HOK human oral keratinocytes at 37°C, 5% CO 2 Oral keratinocyte culture medium was used in the incubator. When the cell confluency rate was 85-90%, the culture medium was removed, washed twice with phosphate buffer solution with a pH value of 7.2, and the washing solution was discarded; the concentration of addition was 0.25% (w / v) trypsin solution for monolayer incub...

Embodiment 3

[0078] A method for detecting the influence of gum-based chewing tobacco on the activity of superoxide dismutase in cells, comprising the following steps:

[0079] Step (1), sample pretreatment: add the gum-based chewing tobacco product to the oral keratinocyte culture medium at 37°C, and the volume ratio of the gum-based chewing tobacco product to the oral keratinocyte culture medium is 1g: 10mL; Then grind for 10 minutes in a constant temperature water bath at 37°C with a grinder, then filter, add serum to the filtrate, so that the final volume percentage concentration of serum is 10%, and obtain the test solution;

[0080] Step (2), preparation of single cell suspension: HOK human oral keratinocytes at 37°C, 5% CO 2 Oral keratinocyte culture medium was used in the incubator. When the cell confluency rate was 84-87%, the medium was removed, washed twice with phosphate buffer solution with a pH value of 7.2, and the washing solution was discarded; the concentration was 0.25% ...

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Abstract

The invention relates to a method for detecting influences of gum based chewing tobacco on cellular superoxide dismutase activity, belonging to the technical field of biological effect evaluation of tobacco and tobacco products. The method comprises the following nine steps: sample pretreatment, preparation of single-cell suspension, cell concentration calculation of the single-cell suspension, test substance exposure, cell collection, sample preparation, sample determination and superoxide dismutase activity calculation. According to the method disclosed by the invention, the fine dissolutionmanner, exposure pathway and action manner of the gum based chewing tobacco are comprehensively surveyed, the sample pretreatment method for the gum based chewing tobacco is established, and effective sample pretreatment, optimized setting of the detection dose and accurate selection of target cells can be realized, so that the influences of the gum based chewing tobacco on the cellular superoxide dismutase activity can be accurately and effectively detected by the method.

Description

technical field [0001] The invention belongs to the technical field of biological effect evaluation of tobacco and tobacco products, and in particular relates to a method for detecting the influence of gum-based chewing tobacco on the enzyme activity of superoxide dismutase in cells. Background technique [0002] As people pay more and more attention to health, higher requirements are put forward for tobacco products, not only to have a better taste, but also to reduce the adverse feelings of the human body as much as possible. Snus products avoid the harm caused by the complex mixture produced by the combustion of traditional cigarette products. At this stage, it has become one of the new directions for foreign tobacco companies to turn from traditional cigarette products. According to the components and morphological characteristics of the product, snus products are divided into traditional Snus-type snus containing tobacco leaf raw materials and new-type snus without toba...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/00G01N33/573G01N33/50G01N33/535G01N33/569
CPCG01N33/00G01N33/5044G01N33/535G01N33/56966G01N33/573G01N2033/0088G01N2333/902
Inventor 管莹高茜朱洲海李雪梅夭建华米其利徐玉琼陆舍铭唐萍
Owner CHINA TOBACCO YUNNAN IND
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