Method for detecting influence of gum-based type tobacco product on human oral cavity in vitro micronucleus

Abstract

The invention discloses a method for detecting the influence of a gum-based type tobacco product on the human oral cavity in vitro micronucleus. The method comprises: (1) placing a gum-based type tobacco product to be detected in liquid nitrogen, and freezing; (2) immediately crushing the frozen gum-based type tobacco product by using a crusher; (3) re-freezing; (4) soaking the crushed gum-based type tobacco product by using artificial saliva; (5) carrying out centrifugal separation on the extraction solution; (6) determining the supernatant through an osmotic pressure determining instrument, and adjusting the osmotic pressure to achieve the same osmotic pressure of a cell culture medium; (7) determining the nicotine content in the extraction solution; (8) culturing human oral mucosal epithelial cells as the detection object; (9) adding the extraction solution to the cultured human oral mucosal epithelial cells; and (10) after the incubation, detecting the cell in vitro micronucleus condition by using 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 the micronucleus of human oral cavity cells in vitro. 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 human oral micronuclei in vitro to evaluate its safety. [0003] The existing safety evaluation methods of tobacco products are mainly aimed at traditional cigaret...

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, and their contents can be dissolved by ordinary soaking methods. Due to the existence of gum base, most of the contents of gum-based tobacco products can be dissolved by direct soaking methods. Contents are difficult to dissolve, making accurate biosafety testing impossible

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