CN110954688B - Method for determining bioavailability of nickel in soil and food for mice - Google Patents
Method for determining bioavailability of nickel in soil and food for mice Download PDFInfo
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- CN110954688B CN110954688B CN201911245086.3A CN201911245086A CN110954688B CN 110954688 B CN110954688 B CN 110954688B CN 201911245086 A CN201911245086 A CN 201911245086A CN 110954688 B CN110954688 B CN 110954688B
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- G—PHYSICS
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- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
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- G01N33/502—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics for testing non-proliferative effects
- G01N33/5038—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics for testing non-proliferative effects involving detection of metabolites per se
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/62—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating the ionisation of gases, e.g. aerosols; by investigating electric discharges, e.g. emission of cathode
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Abstract
The invention discloses a method for determining the mouse biological effectiveness of nickel in soil and food, which comprises the specific steps of preparing food containing nickel and feeding a mouse, collecting urine of the mouse, and finally obtaining the mouse relative biological effectiveness of the nickel in the soil and food.
Description
Technical Field
The invention relates to the field of environmental science, in particular to a method for determining the bioavailability of nickel in soil and food in mice.
Background
Evaluation of the human health risk of nickel in soils and food products is typically assessed using the biological effectiveness of nickel. The human bioavailability of nickel is the ratio of the amount of nickel absorbed by the human body to the total intake of nickel, and the ratio is less than or equal to 100 percent. The method for measuring the biological effectiveness is divided into an animal living body experiment and an in-vitro simulation experiment. In vitro simulation experiments have the advantages of simplicity and convenience, but the results are usually large in error and difficult to simulate the real conditions of human bodies. The animal living body experiment period is long, the operation is complex, but the biological effectiveness of the animal body to the pollutants can be accurately reflected, so that the related human health risk evaluation is carried out. Mice are one of the most commonly used models of animal in vivo experiments. At present, if the conventional mouse experimental method is used for measuring the mouse biological effectiveness of nickel in soil and food, namely, the target organs which finally indicate nickel accumulation are determined as the liver and the kidney, the experiment shows that the nickel cannot be accumulated in the liver and the kidney of the mouse, and the mouse biological effectiveness cannot be measured.
Disclosure of Invention
The present invention aims to provide a method for determining the mouse bioavailability of nickel in soil and food, so as to solve the problems in the background technology.
In order to achieve the purpose, the invention provides the following technical scheme:
a method for determining the mouse bioavailability of nickel in soil and food comprises the following specific steps:
(1) adding a reference substance of soluble nickel into food with low nickel concentration, so as to be convenient for making a standard curve of nickel within a certain concentration range;
(2) grouping mice according to the foods with different concentration ranges in the step (1), feeding a plurality of mice in each group with the mouse food containing the nickel element prepared in the step (1) for 7 days, and then performing 1 day fasting treatment; during 8 days, mice were housed individually in metabolic cages, provided sufficient normal mouse food and Mili-Q water, and urine was isolated and collected;
(3) centrifuging the urine samples collected in step (2) in a centrifuge of 3000r/min for 10min, filtering with 0.45 μm filter membrane, and adding 0.1mol HNO3After diluting by 10 times, analyzing the content of nickel in the urine by ICP-MS;
(4) the consumption of the diet is the difference between the total mass of the diet and the residual amount at the end of the exposure, and the nickel content in the urine is calculated by the product of the concentration and the volume of the nickel in the urine;
the total dose of nickel exposure is the product of the concentration of nickel in the provided rat food and the dietary consumption;
obtaining a urine correction factor:
(5) for the soil to be detected: adding the soil to be measured into food with low nickel concentration at a concentration of 5% to prepare rat food which is easy to be eaten by mice;
directly crushing the food to be measured to prepare rat food which is easy to be eaten by a mouse aiming at the food to be measured;
then calculating the urine dose correction factor of the soil or food to be detected according to the method of the steps (2) to (4), and finally calculating the relative bioavailability of the nickel mouse according to the following formula:
as a further scheme of the invention: the reference substance in the step (1) is Ni Kg in an amount of 0, 0.5, 1.0, 1.5 and 2.5mg-1Is configured.
As a further scheme of the invention: the reference substance in the step (1) is nickel sulfate.
As a further scheme of the invention: and (3) adopting Balb/c for the mice in the step (2), wherein the weight of the mice is 16 g.
Compared with the prior art, the invention has the beneficial effects that:
the method is improved and innovated on the basis of a conventional experimental method, and a method capable of accurately and effectively measuring the bioavailability of nickel in soil and food is designed, namely, the end point of nickel accumulation is positioned in urine, the amount of nickel absorbed by a mouse is obtained by collecting the urine of the mouse and measuring the content of nickel in the urine, and therefore the relative bioavailability of the nickel in the mouse is calculated; is helpful for carrying out systematic evaluation on the nickel content of local crops.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
In the embodiment of the invention, a method for determining the bioavailability of nickel in soil and food by a mouse comprises the following specific steps:
(1) adding soluble nickel reference substance (nickel sulfate, NiSO4) into food with low nickel concentration to facilitate preparation of Ni Kg with certain concentration range (0, 0.5, 1.0, 1.5 and 2.5mg Ni-1) A standard curve of (a);
(2) grouping mice according to the foods with different concentration ranges in the step (1), feeding a plurality of mice (Balb/c, weight-16 g) in each group by using the mouse food containing the nickel element prepared in the step (1), and exposing for 7 days (namely feeding for 7 days, and then performing fasting treatment for 1 day); during 8 days, mice were individually housed in metabolic cages (three samples per concentration of mice fed), provided sufficient normal mouse food and Mili-Q water, and urine was isolated and collected;
(3) centrifuging the urine samples collected in step (2) in a centrifuge of 3000r/min for 10min, filtering with 0.45 μm filter membrane, and filtering with 0.1mol filter membraneHNO3After diluting by 10 times, analyzing the content of nickel in the urine by ICP-MS;
(4) the consumption of the diet is the difference between the total mass of the diet and the residual amount at the end of the exposure, and the nickel content in the urine is calculated by the product of the concentration and the volume of the nickel in the urine;
the total dose of nickel exposure is the product of the concentration of nickel in the provided rat food and the dietary consumption;
obtaining a urine correction factor:
(5) for the soil to be detected: adding the soil to be measured into food with low nickel concentration at a concentration of 5% to prepare rat food which is easy to be eaten by mice;
directly crushing the food to be measured to prepare rat food which is easy to be eaten by a mouse aiming at the food to be measured;
then calculating the urine dose correction factor of the soil or food to be detected according to the method of the steps (2) to (4), and finally calculating the relative bioavailability of the nickel mouse according to the following formula:
it will be evident to those skilled in the art that the invention is not limited to the details of the foregoing illustrative embodiments, and that the present invention may be embodied in other specific forms without departing from the spirit or essential attributes thereof. The present embodiments are therefore to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein.
Furthermore, it should be understood that although the present description refers to embodiments, not every embodiment may contain only a single embodiment, and such description is for clarity only, and those skilled in the art should integrate the description, and the embodiments may be combined as appropriate to form other embodiments understood by those skilled in the art.
Claims (4)
1. A method for determining the mouse bioavailability of nickel in soil and food is characterized by comprising the following specific steps:
(1) adding a reference substance of soluble nickel into food with low nickel concentration, so as to be convenient for making a standard curve of nickel within a certain concentration range;
(2) grouping mice according to the foods with different concentration ranges in the step (1), feeding a plurality of mice in each group with the mouse food containing the nickel element prepared in the step (1) for 7 days, and then performing 1 day fasting treatment; during 8 days, mice were housed individually in metabolic cages, provided sufficient normal mouse food and Mili-Q water, and urine was isolated and collected;
(3) centrifuging the urine samples collected in step (2) in a centrifuge of 3000r/min for 10min, filtering with 0.45 μm filter membrane, and adding 0.1mol HNO3After diluting by 10 times, analyzing the content of nickel in the urine by ICP-MS;
(4) the consumption of the diet is the difference between the total mass of the diet and the residual amount at the end of the exposure, and the nickel content in the urine is calculated by the product of the concentration and the volume of the nickel in the urine;
the total dose of nickel exposure is the product of the concentration of nickel in the provided rat food and the dietary consumption;
obtaining a urine correction factor:
in a reference substance
Preparing a standard reference curve according to the content of nickel in the urine, the total exposed nickel dose and the urine correction factor;
(5) for the soil to be detected: adding the soil to be measured into food with low nickel concentration at a concentration of 5% to prepare rat food which is easy to be eaten by mice;
directly crushing the food to be measured to prepare rat food which is easy to be eaten by a mouse aiming at the food to be measured;
then calculating a urine dose correction factor of the soil or food to be detected according to the method of the steps (2) to (4), and finally calculating the relative bioavailability of the nickel mouse according to the following formula:
2. the method for determining mouse bioavailability of nickel in soil and food according to claim 1, wherein the reference substance in step (1) is Ni Kg in the range of 0, 0.5, 1.0, 1.5 and 2.5mg Ni-1Is configured.
3. The method for determining mouse bioavailability of nickel in soil and food according to claim 1 or 2, wherein the reference substance in step (1) is nickel sulfate.
4. The method for determining the bioavailability of nickel in soil and food for mice according to claim 1, wherein the mice in step (2) adopt Balb/c and have a body weight of 16 g.
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