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Establishment method of heavy metal oral bioaccessibility detection system

A technology for establishing methods and heavy metals, applied in biological testing, measuring devices, material inspection products, etc., can solve the problem of affecting the accuracy of biological accessibility detection and health risk assessment, difficult to reflect the absorption of human body, and ignoring the impact of chemical transformation. and other problems, to achieve the effects of predicting toxic effects, detecting biological accessibility, and controllable experimental conditions.

Active Publication Date: 2020-08-07
SUN YAT SEN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, in vitro cell models often use the method of directly infecting the Caco-2 cell model or other intestinal tissue-derived cells with the chemical to be tested to simulate the intestinal absorption and transformation process of the chemical. Compared with the in vivo study, this method is simple to operate and low in cost. , It can preliminarily evaluate the impact of human exposure to chemicals in a relatively short period of time, but there are obvious defects: (1) The Caco-2 cell model is only composed of a single layer of small intestinal epithelial cells (2) The method of directly using chemicals for poisoning ignores the influence of the human body’s digestive process on chemical conversion, especially when evaluating unstable chemicals in food and drinking water, the digestive process cannot ignore
This method only determines the bioaccessibility through the in vitro digestion model, without considering the absorption situation, and it is difficult to reflect the actual absorption situation of the human body, thus affecting the accuracy of bioaccessibility detection and health risk assessment

Method used

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  • Establishment method of heavy metal oral bioaccessibility detection system
  • Establishment method of heavy metal oral bioaccessibility detection system
  • Establishment method of heavy metal oral bioaccessibility detection system

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Experimental program
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Embodiment 1

[0037] Embodiment 1 Method for establishing a detection system for oral bioaccessibility of heavy metals

[0038] A method for establishing a heavy metal oral bioaccessibility detection system is provided, comprising the following steps:

[0039] 1.1 Construct a human in vitro gastrointestinal digestion model and a 3D model of human small intestinal epithelial cells;

[0040] The in vitro digestion model is established based on the static in vitro digestion InfoGest2.0 method. Before the start, it is necessary to measure the activity of pepsin and trypsin, the concentration of bile salt and the volume of acid and alkali required to adjust the pH to a specific value to control the batch difference between experiments and improve Experimental repeatability. Prepare 1.25× simulated saliva fluids (SSF) stock solution, (1× simulated saliva contains 15.1mM potassium chloride, 3.7mM potassium dihydrogen phosphate, 13.6mM sodium bicarbonate, 0.15mM magnesium chloride hexahydrate, 0.0...

Embodiment 2

[0056] The establishment method of embodiment double heavy metal oral toxicity detection system

[0057] A method for establishing a heavy metal oral toxicity detection system is provided, comprising the following steps:

[0058] 1.1 Construct in vitro digestion model and 3D model of human intestinal epithelial cells;

[0059] 1.2 Apply different concentrations of heavy metals to the in vitro digestion model to simulate the human digestion process, and collect heavy metal-containing digestive juice;

[0060] 1.3 After applying the digestive juice containing heavy metals to the human small intestinal epithelial cell in vitro absorption model, culture it for 4-24 hours to simulate the small intestinal absorption process, and measure the change curve reflecting the degree of toxic damage of the cell model with the applied concentration of heavy metals;

[0061] 1.4 Track the concentration changes of heavy metals / digested products with heavy metal characteristics at each stage of...

Embodiment 3

[0073] The quality control involved in the system application process of embodiment three

[0074] The invention performs quality control by calculating the recovery rate in the digestion / absorption stage to ensure the accuracy of the result. In the digestion stage, the concentration of heavy metals in the supernatant (digestion) obtained by centrifugation after digestion and the insoluble precipitate (chyme) after centrifugation is measured. The ratio of the total amount of heavy metals contained in the two to the content of heavy metals in the food sample is the recovery rate of the digestion stage. The recovery rate at this stage is required to be in the range of 0.9-1.1; the concentration of heavy metals in AP, BL and cells are measured respectively during the intestinal absorption stage, and the ratio of the sum of the three to the heavy metal content of AP added to the digestive juice is the recovery rate in the intestinal absorption stage. Stage recoveries ranged from 0...

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Abstract

The invention provides an establishment method of a heavy metal oral bioaccessibility detection system. The establishment method comprises the following steps: 1) respectively establishing a human body external gastrointestinal digestion model and a human small intestine epithelial cell 3D model; 2) respectively applying substances containing different concentrations of heavy metals to the human body external gastrointestinal digestion model to simulate the gastrointestinal digestion process; 3) respectively applying the digestive juice containing the heavy metals to the human small intestineepithelial cell 3D model, and performing culturing for a preset time to simulate a small intestine absorption process; and 4) tracking changes of heavy metal concentrations and valence states in the human body external gastrointestinal digestion process and the small intestine epithelial cell absorption process, calculating oral biological accessibility of heavy metals, and establishing a method for predicting exposure risks based on the biological accessibility. The invention belongs to the technical field of food safety detection and evaluation, is suitable for detecting oral biological accessibility of heavy metals, and is also suitable for potential toxicity effect detection and exposure risk evaluation.

Description

technical field [0001] The invention belongs to the technical field of food safety detection and evaluation, and in particular relates to a method for establishing a heavy metal oral bioaccessibility and toxicity detection system. Background technique [0002] Affected by environmental pollution and other factors, safety issues of food and drinking water sources occur from time to time. The incidence of diseases caused by oral intake of contaminated food / drinking water is increasing day by day, and has occupied the second place in the list of the total incidence of various diseases, and has become a safety and health issue that is currently focused on. Heavy metals, as an important chemical pollutant in nature, once they enter the body, are difficult to be discharged naturally or completely removed. The harm to the body has the characteristics of multi-system, multi-organ and irreversible, which seriously affects human health and safety. Therefore, it is imperative to condu...

Claims

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Application Information

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IPC IPC(8): G01N33/50
CPCG01N33/5014G01N33/5044
Inventor 陈雯王紫微张睿李道传陈丽萍
Owner SUN YAT SEN UNIV
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