Method for evaluating biological safety of colored plastic based on zebra fish

Abstract

The invention discloses a method for evaluating the biological safety of colored plastics based on zebra fish. The method comprises the following steps: acquiring the half lethal concentration of zebra fish embryos of the colored plastics; determining the concentration of the colored plastic in the water body to be evaluated and performing secondary judgment; according to the method, the safety of the colored plastic in the water body can be subjected to risk assessment through zebra fish embryo indexes, so that whether the colored plastic in the water body generates toxicity to the ecological environment and human health or not can be accurately predicted, subsequent assessment and management and control of the colored plastic are facilitated, and the purposes of high efficiency and environmental protection are achieved.

Description

Technical Field

[0001] The present invention relates to the technical field of toxicity evaluation of colored plastics, and in particular to a method for evaluating the biosafety of colored plastics based on zebrafish. Background Art

[0002] Plastics are made from monomers of petroleum and natural gas, and have excellent characteristics such as low cost, high toughness, corrosion resistance, and water resistance. With the rapid development of social economy, industry and agriculture, plastic products are widely used in packaging, automobiles, construction, electronics, information technology, and communications. The global demand for plastics has gradually expanded, and the output of plastic products has shown exponential growth.

[0003] Inorganic pigments are widely used in the industry due to their thermal stability, high chemical resistance, strong coloring ability and high brightness. Generally speaking, due to the low solubility of inorganic pigments, the leaching concentration of heavy metals is low and the ecological health risk is low. However, recent studies have shown that when pigments are added to plastics, as the plastics break into microplastics in the environment, the leaching concentration increases significantly, and the ecological and human health risks caused cannot be ignored.

[0004] As an important aquatic model organism, zebrafish is highly adaptable and easy to raise. In addition, it is transparent during the embryonic stage, making it easier to observe its appearance. Currently, the whole genome sequencing of zebrafish has been completed, and it has been found that zebrafish and human genes have about 70% homology, and 84% of human disease-related genes can find homologous genes in the zebrafish genome.

[0005] Exposure to microplastics can cause endocrine disorders, intestinal damage, reduced nutrient intake, behavioral changes, and even increased mortality in organisms. Studies have shown that microplastics can cause oxidative stress and cytotoxicity in organisms. After Lu et al. exposed zebrafish to microplastics for seven days, they found that microplastics with particle sizes of 5 μm and 70 nm could cause inflammation and lipid accumulation in fish liver, and induce a significant increase in superoxide dismutase and catalase activity, and zebrafish showed oxidative stress. In in vitro experiments by Geiser et al., lung macrophages and red blood cells can ingest PS fluorescent microspheres (1, 0.2, 0.078 μm) by diffusion or adhesion, which means that plastic microspheres can directly enter proteins, organelles, and even DNA in cells, greatly enhancing their toxic potential. In addition, the presence of microplastics can also lead to gastrointestinal toxicity, directly or indirectly interfering with metabolism and energy changes. Jin et al. found that the mouse intestine was affected by PS microplastics, reducing intestinal mucus secretion and impairing intestinal barrier function. In addition, the intestinal flora also changed. Exposure to microplastics increased the anaerobic enzyme lactate dehydrogenase (LDH) in fish and regulated related RNA. In the liver of mice, microplastics can lead to reduced ATP levels and reduced lipid metabolism.

[0006] Due to the unique material, hydrophobicity and high surface area to volume ratio of microplastics, they can be used as carriers to transport adsorbed compounds. Their combined exposure with pollutants such as heavy metals, polycyclic aromatic hydrocarbons, and residual drugs may lead to enhanced toxicity and cause unpredictable ecological health risks.

[0007] However, few studies have focused on the toxicity of heavy metal pigments in colored plastics, and there are still many gaps in their related biological toxicity, which deserves more in-depth research. Summary of the invention

[0008] In order to solve the above problems, the present invention provides a method for evaluating the biosafety of colored plastics based on zebrafish.

[0009] A method for evaluating the biosafety of colored plastics based on zebrafish comprises the following steps:

[0010] Obtaining the zebrafish embryo median lethal concentration of colored plastic; wherein the colored plastic is plastic that has been injection molded with a heavy metal dye;

[0011] Determine the concentration of colored plastics in the water body to be assessed;

[0012] If the concentration of colored plastics in the water to be assessed is greater than or equal to the median lethal concentration of zebrafish embryos of the colored plastics, the water to be assessed is judged to be unsafe; conversely, if the concentration of colored plastics in the water to be assessed is less than the median lethal concentration of zebrafish embryos of the colored plastics, a secondary judgment is made;

[0013] The secondary judgment includes:

[0014] Determine the maximum release concentration of heavy metals from colored plastics during the evaluation period;

[0015] Taking the water body to be evaluated with the maximum release concentration of heavy metals in the colored plastic, cultivating zebrafish embryos, obtaining the experimental value of the zebrafish embryo index, and comparing the significant difference with the standard value of the zebrafish embryo index to determine whether the experimental value of the zebrafish embryo index exceeds the standard;

[0016] If the standard is exceeded, it means that the water body to be assessed is unsafe within the assessment period; if the standard is not exceeded, it means that the water body to be assessed is safe within the assessment period.

[0017] Note: The above method first roughly judges whether the water body is seriously polluted based on the concentration of colored plastics. If the concentration of colored plastics itself is high, it means that it is already highly toxic and there is no need for secondary detailed judgment. If the concentration of colored plastics is temporarily small, but over time, it may release heavy metals and other harmful substances, causing long-term and unpredictable pollution. The above secondary judgment method can be used to make more accurate predictions and judgments, so as to improve the accuracy of the biosafety evaluation of colored plastics in water bodies.

[0018] Further, the method for determining the maximum release concentration of heavy metals in colored plastics within the evaluation time includes:

[0019] Determine the evaluation time, and calculate the maximum release concentration of heavy metals in the colored plastic based on the evaluation time; wherein the concentration release range of the heavy metals in the colored plastic is related to the illumination duration and the illumination intensity.

[0020] Note: Through the above method, the maximum concentration of non-ferrous metals that can be released at the time point to be evaluated can be determined, and the study found that the amount of release is related to the parameters of light. Therefore, the release concentration of heavy metals in colored plastics can be predicted through light parameters.

[0021] Further, the calculation of the maximum release concentration of heavy metals in colored plastics based on the evaluation time includes:

[0022] Conducting a correlation analysis on the concentration of the colored plastic in the water body to be evaluated and the concentration release range of the heavy metals in the colored plastic under multiple sets of preset light durations and light intensities, and obtaining multiple release coefficients under multiple sets of preset light durations and light intensities;

[0023] The release coefficient when the light intensity is maximum under the evaluation time is determined, and the maximum release concentration of heavy metals in the colored plastic is calculated using the release coefficient when the light intensity is maximum and the concentration of the colored plastic.

[0024] Furthermore, the method for determining the range of the concentration of heavy metals released in the colored plastic is as follows:

[0025] Under multiple sets of preset illumination durations and illumination intensities, and within the evaluation time, the concentrations of heavy metals in multiple sets of preset concentrations of colored plastics are regularly tested to obtain the range of concentration release of heavy metals in the colored plastics.

[0026] Note: The above calculation method can obtain the release coefficient, which can be used to calculate the release concentration of heavy metals in colored plastics.

[0027] Furthermore, the release concentration of heavy metals in the colored plastic is calculated by a prediction model;

[0028] The prediction model satisfies:

[0029] C=k·I a ·t b

[0030] Wherein, C is the release concentration of heavy metals in colored plastics, in mg / L, k is the characteristic constant, I is the light intensity, a is the release coefficient of light intensity, t is the light duration, and b is the release coefficient of light duration.

[0031] Note: The above formula is the relationship between the release concentration of heavy metals in colored plastics, illumination parameters and release coefficient. Through this relationship, the release concentration of heavy metals in colored plastics can be obtained through model calculation under known illumination parameters without on-site testing. At the same time, the release concentration of heavy metals in non-ferrous metals can be predicted to improve the accuracy of colored plastic assessment.

[0032] Furthermore, the characteristic constant k is related to the characteristics of the colored plastic, the pH value, temperature and water flow velocity of the water body to be evaluated.

[0033] Note: Since different plastic components have different release effects on different heavy metals, and the release process will be affected by the water body itself, the characteristic constants are also different. By obtaining appropriate characteristic constants, the calculation effect of the prediction model can be improved.

[0034] Furthermore, the method of obtaining the zebrafish embryo median lethal concentration of the colored plastic includes:

[0035] Obtaining colored plastic aqueous solutions at multiple concentrations;

[0036] Toxicity tests were conducted using zebrafish embryos, and the zebrafish embryo median lethal concentration was obtained.

[0037] Furthermore, the zebrafish embryo indicators include: one or more of mortality rate, deformity rate, heart rate, developmental delay, number of autonomous movements and embryo oxidative stress indicators.

[0038] Note: The above indicators can more accurately characterize the impact of the water to be tested on zebrafish embryos, and the biological characteristics of zebrafish to toxicity are similar to those of humans, which can more accurately reflect whether the water to be tested is harmful to the human body.

[0039] Further, judging whether the cultured zebrafish embryos exceed the standard according to the experimental values ​​of the zebrafish embryo indicators includes:

[0040] Take water bodies that are not polluted by colored plastics, cultivate zebrafish embryos, and obtain standard values ​​of zebrafish embryo indicators;

[0041] Compare the experimental values ​​of zebrafish embryo indicators with the standard values ​​for significant differences;

[0042] If there is a significant difference, it means that the experimental value of the zebrafish embryo index exceeds the standard. If there is no significant difference, it means that the experimental value of the zebrafish embryo index does not exceed the standard.

[0043] Note: Through the above significant differences, it is possible to scientifically measure whether the water to be tested has a strong toxic effect on zebrafish embryos.

[0044] The beneficial effects of the present invention are:

[0045] The purpose of the present invention is to propose a method for evaluating the biosafety of colored plastics based on zebrafish to solve the problem of lack of evaluation of the biosafety of colored plastics. The method of the present invention first roughly judges whether the water body is seriously polluted based on the concentration of colored plastics. If the concentration of colored plastics itself is relatively high, it means that it has strong toxicity and there is no need to perform secondary subdivision judgment. If the concentration of colored plastics is temporarily small, it may release harmful substances such as heavy metals over time, causing long-term and unpredictable pollution. Through the above-mentioned secondary judgment method, it is possible to make more accurate predictions and judgments to improve the accuracy of the biosafety evaluation of colored plastics in water bodies. At the same time, through the setting of the prediction model, it is possible to perform model calculations under known lighting parameters to obtain the release concentration of heavy metals in colored plastics without on-site detection. At the same time, the release concentration of heavy metals in non-ferrous metals can be predicted to improve the accuracy of the evaluation of colored plastics. BRIEF DESCRIPTION OF THE DRAWINGS

[0046] Figure 1 is a data graph of the mortality rate, hatching rate, and deformity rate of zebrafish in an embodiment of the present invention;

[0047] Figure 2is a heavy metal detection data diagram of colored plastics in an embodiment of the present invention;

[0048] Figure 3 is a schematic diagram of developmental indicators of zebrafish embryos in an embodiment of the present invention;

[0049] Figure 4 is a schematic diagram of oxidative stress levels in zebrafish according to an embodiment of the present invention;

[0050] Figure 5 is a data graph of zebrafish oxidative stress levels in an embodiment of the present invention;

[0051] Figure 6 is an example data diagram of a prediction model in an embodiment of the present invention;

[0052] Figure 7 It is a schematic diagram of the processing results of each step of the present invention on zebrafish. DETAILED DESCRIPTION

[0053] In order to further illustrate the method adopted by the present invention and the effect achieved, the technical solution of the present invention will be clearly and completely described in combination with experiments below.

[0054] The method provided by the embodiment of the present invention relates to the technical field of toxicity assessment of colored plastics. The safety of colored plastics in water bodies can be risk assessed through zebrafish embryo indicators, so as to more accurately predict whether the colored plastics in water bodies will be toxic to the ecological environment and human health, thereby facilitating subsequent evaluation and control of colored plastics and achieving the purpose of high efficiency and environmental protection.

[0055] In the process of studying the toxicity of plastics in water bodies, the existing technology usually directly uses various indicators to conduct tests, and evaluates whether the plastics in the water body are safe by judging whether these indicators exceed the standard under the concentration of plastics. However, this method is not very accurate in the evaluation process of colored plastics, because colored plastics dyed with heavy metal pigments will release heavy metals into the water body, and the existing related methods are difficult to measure the subsequent release of heavy metals and the possible pollution. At the same time, the release of heavy metals in colored plastics does not exceed the standard according to the heavy metal standard, but harm may have been caused during the release process. Therefore, a more accurate evaluation method is needed to measure the hazards of colored plastics.

[0056] Based on the above, the present application provides a method for evaluating the biosafety of colored plastics based on zebrafish, comprising the following steps:

[0057] S1. Obtaining the zebrafish embryo median lethal concentration of colored plastic; wherein the colored plastic is a plastic that has been injection molded with a heavy metal dye;

[0058] The zebrafish embryo median lethal concentration of the colored plastics includes:

[0059] S1-1, obtaining colored plastic aqueous solutions of multiple concentrations;

[0060] For example, colored plastics dyed with cadmium red pigment are used to prepare colored plastic solutions with multiple concentrations, and then zebrafish embryos are used for toxicity tests to obtain the LC50 value of zebrafish embryos; and the exposure concentration is determined based on the LC50 and the environmental concentration;

[0061] The above-mentioned multiple concentration groups are respectively the Control group with illumination, the inorganic pigment group with illumination, the plastic group with illumination, the color plastic group with illumination, and the Control group without illumination, the inorganic pigment group without illumination, the plastic group without illumination, and the color plastic group without illumination, for a total of 8 groups.

[0062] S1-2: Toxicity test was conducted using zebrafish embryos to obtain the zebrafish embryo median lethal concentration.

[0063] S2. Determine the concentration of colored plastics in the water body to be assessed;

[0064] Exemplarily, the exposure concentration may be set to 1 mg / L.

[0065] S3. Determine the safety of water bodies

[0066] S3-1. If the concentration of colored plastics in the water body to be assessed is greater than or equal to the median lethal concentration of zebrafish embryos of the colored plastics, the water body to be assessed is deemed unsafe;

[0067] S3-2. On the contrary, if the concentration of colored plastics in the water body to be evaluated is lower than the median lethal concentration of the colored plastics for zebrafish embryos, a secondary judgment is made;

[0068] The secondary judgment includes:

[0069] S3-21. Determine the maximum release concentration of heavy metals in colored plastics during the assessment period;

[0070] The method for determining the maximum release concentration of heavy metals in colored plastics within the evaluation time includes:

[0071] Determine the assessment time; the assessment time refers to a time in the future when the water body needs to be assessed. For example, if the assessment time is 3 days, it means that the water body needs to be assessed 3 days later;

[0072] Based on the evaluation time, the maximum release concentration of heavy metals in colored plastics is calculated; wherein, the concentration release range of heavy metals in the colored plastics is related to the illumination duration and the illumination intensity.

[0073] The calculation of the maximum release concentration of heavy metals in colored plastics based on the evaluation time includes:

[0074] Conducting a correlation analysis on the concentration of the colored plastic in the water body to be evaluated and the concentration release range of the heavy metals in the colored plastic under multiple sets of preset light durations and light intensities, and obtaining multiple release coefficients under multiple sets of preset light durations and light intensities;

[0075] The release coefficient when the light intensity is maximum under the evaluation time is determined, and the maximum release concentration of heavy metals in the colored plastic is calculated using the release coefficient when the light intensity is maximum and the concentration of the colored plastic.

[0076] Wherein, the range of determining the concentration release of heavy metals in colored plastics includes:

[0077] Under multiple sets of preset illumination durations and illumination intensities, and within the evaluation time, the concentration release of heavy metals in multiple sets of preset concentrations of colored plastics are regularly tested to obtain the range of the concentration release of heavy metals in the colored plastics.

[0078] The release concentration of heavy metals in the colored plastic is calculated by a prediction model;

[0079] The prediction model satisfies:

[0080] C=k·I a ·t b

[0081] In the formula, C is the release concentration of heavy metals in colored plastics, the unit is mg / L, k is the characteristic constant, the value range is 0.95~1.7, I is the light intensity, a is the release coefficient of light intensity, the value range is 0.6~0.8, t is the light duration, b is the release coefficient of light duration, the value range is 0.7~0.9.

[0082] The characteristic constant k is related to the characteristics of the colored plastic, the pH value, temperature and water flow velocity of the water body to be evaluated. Further, when the characteristic constant k satisfies: In the formula, P is the plastic parameter (the value range is 1 to 2, for example, the value range of polyethylene is 1 to 1.5, the value range of polypropylene is 1.2 to 1.8, the value range of polyvinyl chloride is 1.5 to 2, and the value range of polystyrene is 1 to 1.3), H is the pH value of the water body to be evaluated, T is the temperature, ℃, and V is the water flow velocity, m / s).

[0083] For example, as shown in Table 1, using 1 mg / L CdSSe-PS, the illumination intensity I is constant at 1.627 ± 0.138 mW / cm 2 , under different light durations, detect Cd 2+ The release concentration is as follows, and the fitting curve is as follows Figure 6 As shown, y = 0.3615x + 1.8480 (in the figure, y is Cd2+ concentration, x is the illumination duration), which is consistent with the above prediction model.

[0084] Table 1 Cd under different light durations 2+ Concentration results table

[0085] t is the duration of illumination (h) <![CDATA[Cd 2+ Concentration (ppb)]]> 3 2.86875 5 3.96549 7 4.77542

[0086] S3-22, taking the water body to be evaluated with the maximum release concentration of heavy metals in the colored plastic, cultivating zebrafish embryos, obtaining the experimental value of the zebrafish embryo index, and judging whether the cultivated zebrafish embryos exceed the standard according to the experimental value of the zebrafish embryo index;

[0087] If the standard is exceeded, it means that the water body to be assessed is unsafe within the assessment period; if the standard is not exceeded, it means that the water body to be assessed is safe within the assessment period.

[0088] The method of judging whether the cultivated zebrafish embryos exceed the standard according to the experimental values ​​of the zebrafish embryo indicators comprises:

[0089] Take water bodies that are not polluted by colored plastics, cultivate zebrafish embryos, and obtain standard values ​​of zebrafish embryo indicators;

[0090] Compare the experimental values ​​of zebrafish embryo indicators with the standard values ​​for significant differences;

[0091] If there is a significant difference, it means that the experimental value of the zebrafish embryo index exceeds the standard. If there is no significant difference, it means that the experimental value of the zebrafish embryo index does not exceed the standard.

[0092] Exemplarily, the zebrafish embryo indicators include: one or more of mortality rate, deformity rate, heart rate, developmental delay, number of autonomous movements and embryo oxidative stress indicators;

[0093] Sublethal indicators of zebrafish embryos include: mortality, deformity rate, heart rate, developmental delay and reduced number of autonomous movements. The deformities include yolk sac edema, pericardial edema, spinal curvature, etc.; the developmental delay includes delayed hatching and decreased body length, etc.; embryo oxidative stress indicators include: distribution of reactive oxygen species (ROS) fluorescence intensity, catalase CAT, glutathione (GSH), malondialdehyde (MDA), etc.

[0094] The following are the indicators of zebrafish in this example at a concentration of 1 mg / L of colored plastic:

[0095] like Figure 1 As shown, Figure 1 A in the middle shows the hatching rate, mortality rate and deformity rate of zebrafish after exposure to heavy metal pigments at multiple time points; B shows the hatching rate, mortality rate and deformity rate of zebrafish after exposure to colored plastics. Figure 1The figure includes different concentrations of CdSSe and CdSSe-PS exposure groups, and the hatching rate, mortality rate and deformity rate of zebrafish at 24, 48, 72, 96, 120, 144 and 168 hpf. A: CdSSe pigment exposure group. B: CdSSe-PS particle exposure group. i: hatching rate; ii: mortality rate;

[0096] iii: Deformity rate.

[0097] from Figure 1 It can be seen that compared with the control group, the zebrafish in the exposure group showed delayed hatching, increased mortality and deformity rates, proving that the hatching rate, mortality and deformity rates of zebrafish can quickly characterize water pollution.

[0098] like Figure 2 As shown, Figure 2 are the concentrations of Cd and Se in the control group and the exposed group, A is the concentration of Cd and Se in the solution when there is no zebrafish exposure, showing a linear release, and there is a high correlation between the concentrations of Cd and Se. B and C are the concentrations of Cd and Se in zebrafish tissue samples and exposed water, and the concentrations of Cd and Se in the samples when there is zebrafish exposure. That is, (A) the concentration of Cd and Se in the solution when there is no material exposure. Ai: Cd concentration; A-ii: Selenium concentration; A-iii: Correlation between Cd and Se concentrations. (B&C) Cd and Se concentrations in the solution in zebrafish tissue samples or material exposure groups. i: solution concentration under normal lighting conditions; ii: solution concentration under dark field lighting conditions; iii: zebrafish tissue sample concentration under normal lighting conditions; iv: zebrafish tissue sample concentration under dark field lighting conditions.

[0099] from Figure 2 It can be seen that as the illumination time increases, heavy metal ions in the material are released, and the concentrations of Cd and Se in zebrafish increase, which has a greater impact on zebrafish. By measuring the concentrations of heavy metals in exposed water and organisms, the impact of colored pigments on zebrafish can be reflected more quickly.

[0100] like Figure 3 As shown, Figure 3 The following are typical images of zebrafish in the lateral position, including typical images of zebrafish in the lateral position at 24, 120, 144 and 168 hpf. It can be seen that with the increase in exposure time, the body shape of zebrafish gradually bends in the light exposure, accompanied by changes in basic developmental parameters. This shows that under different concentrations of colored plastics, different morphological characteristics of zebrafish can be seen, proving that the morphology of zebrafish can be used to characterize the pollution of colored plastics to water bodies.

[0101] like Figure 4 As shown, Figure 4These are the relevant developmental parameters of zebrafish and the basic developmental parameters of zebrafish. A: Yolk sac area at 24hpf. B: Number of tail flicks at 24hpf. C: Heartbeat at 72hpf. D: Swim bladder area at 168hpf. E: Body curvature at 168hpf. F: Body length at 168hpf.

[0102] It can be seen that the basic developmental indicators of zebrafish are all affected by the materials, which proves that the basic developmental indicators of zebrafish can be used to characterize the pollution of water bodies.

[0103] like Figure 5 As shown, Figure 5 The following are the ROS accumulation diagrams and oxidative stress-related parameter levels after using the DCFH-DA probe. Typical images of ROS generation and oxidative stress-related parameter levels in 168hpf zebrafish larvae exposed to different materials. A: ROS fluorescence distribution diagram after using the DCFH-DA probe; B: ROS fluorescence intensity after using the DCFH-DA probe; C: Glutathione level; D: SOD activity level; E: Malondialdehyde content (percentage of the control group);

[0104] from Figure 5 It can be seen that in the light field exposure group, colored plastics have a greater impact on the oxidative stress level of zebrafish than in the dark field exposure group, proving that under light conditions, colored plastics will have greater biological toxicity to zebrafish, indicating that the oxidative stress level of zebrafish can be used to evaluate water pollution.

[0105] The above-mentioned significant difference method adopts the one-way analysis of variance method in the prior art to clarify the significant differences in embryonic indicators between the blank control group (standard values ​​of zebrafish embryonic indicators) and the experimental group (experimental values ​​of zebrafish embryonic indicators), and evaluate the biosafety of the colored plastic to zebrafish; exemplarily, the significant difference level is expressed as: p < 0.05 or p < .01 or p < 0.001.

Claims

1. A method for evaluating the biosafety of colored plastics based on zebrafish, characterized in that: The following steps are involved: Obtaining the zebrafish embryo median lethal concentration of colored plastic; wherein the colored plastic is plastic that has been injection molded with a heavy metal dye; Determine the concentration of colored plastics in the water body to be assessed; If the concentration of colored plastics in the water to be assessed is greater than or equal to the median lethal concentration of zebrafish embryos of the colored plastics, the water to be assessed is judged to be unsafe; conversely, if the concentration of colored plastics in the water to be assessed is less than the median lethal concentration of zebrafish embryos of the colored plastics, a secondary judgment is made; The secondary judgment includes: Determine the maximum release concentration of heavy metals from colored plastics during the evaluation period; Taking the water body to be evaluated with the maximum release concentration of heavy metals in the colored plastic, cultivating zebrafish embryos, obtaining the experimental value of the zebrafish embryo index, and comparing the significant difference with the standard value of the zebrafish embryo index to determine whether the experimental value of the zebrafish embryo index exceeds the standard; If the standard is exceeded, it means that the water body to be assessed is unsafe within the assessment period; if the standard is not exceeded, it means that the water body to be assessed is safe within the assessment period.

2. A method for evaluating the biosafety of colored plastics based on zebrafish as claimed in claim 1, characterized in that: The method for determining the maximum release concentration of heavy metals in colored plastics within the evaluation time includes: Determine the evaluation time, and calculate the maximum release concentration of heavy metals in the colored plastic based on the evaluation time; wherein the concentration release range of the heavy metals in the colored plastic is related to the illumination duration and the illumination intensity.

3. A method for evaluating the biosafety of colored plastics based on zebrafish as claimed in claim 2, characterized in that: The calculation of the maximum release concentration of heavy metals in colored plastics based on the evaluation time includes: Conducting a correlation analysis on the concentration of the colored plastic in the water body to be evaluated and the concentration release range of the heavy metals in the colored plastic under multiple sets of preset light durations and light intensities, and obtaining multiple release coefficients under multiple sets of preset light durations and light intensities; The release coefficient when the light intensity is maximum under the evaluation time is determined, and the maximum release concentration of heavy metals in the colored plastic is calculated using the release coefficient when the light intensity is maximum and the concentration of the colored plastic.

4. A method for evaluating the biosafety of colored plastics based on zebrafish as claimed in claim 3, characterized in that: The method for determining the range of heavy metal concentration release in the colored plastic is as follows: Under multiple sets of preset illumination durations and illumination intensities, and within the evaluation time, the concentrations of heavy metals in multiple sets of preset concentrations of colored plastics are regularly tested to obtain the range of concentration release of heavy metals in the colored plastics.

5. The method for evaluating the biosafety of colored plastics based on zebrafish as claimed in claim 1, characterized in that: The maximum release concentration of heavy metals in the colored plastic is calculated by a prediction model; The prediction model satisfies: C=k·I a ·t b Wherein, C is the maximum release concentration of heavy metals in colored plastics, in mg / L, k is the characteristic constant, I is the light intensity, a is the release coefficient of light intensity, t is the light duration, and b is the release coefficient of light duration.

6. A method for evaluating the biosafety of colored plastics based on zebrafish as claimed in claim 5, characterized in that: The characteristic constant k is related to the characteristics of the colored plastic, the pH value, temperature and water flow rate of the water body to be evaluated.

7. The method for evaluating the biosafety of colored plastics based on zebrafish as claimed in claim 1, characterized in that: The zebrafish embryo median lethal concentration of the colored plastics includes: Toxicity tests were conducted on colored plastic aqueous solutions of various concentrations using zebrafish embryos to obtain the median lethal concentration for zebrafish embryos.

8. The method for evaluating the biosafety of colored plastics based on zebrafish as claimed in claim 1, characterized in that: The zebrafish embryo indicators include: one or more of mortality rate, deformity rate, heart rate, developmental delay, number of autonomous movements and embryo oxidative stress indicators.

9. The method for evaluating the biosafety of colored plastics based on zebrafish as claimed in claim 1, characterized in that: The method of judging whether the cultivated zebrafish embryos exceed the standard according to the experimental values ​​of the zebrafish embryo indicators comprises: Take water bodies that are not polluted by colored plastics, cultivate zebrafish embryos, and obtain standard values ​​of zebrafish embryo indicators; Compare the experimental values ​​of zebrafish embryo indicators with the standard values ​​for significant differences; If there is a significant difference, it means that the experimental value of the zebrafish embryo index exceeds the standard. If there is no significant difference, it means that the experimental value of the zebrafish embryo index does not exceed the standard.

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