Application of pentachlorophenol as standard reference substance in detecting interference effect of environmental sample on retinoic acid receptor beta

Abstract

The invention relates to the technical field of environment detection, in particular to application of pentachlorophenol as a standard reference substance to detection of interference effect of an environment sample on a retinoic acid receptor beta. The pentachlorophenol toxicity equivalent concentration of the triggering value of the interference effect is 10.77 [mu] g / L. A standard reference compound pentachlorophenol for the in-vitro biological test of the retinoic acid receptor beta interference effect of the water environment sample is screened out, the compound is high in detection rate, has a good linear interval on a dose-effect relation curve of the retinoic acid receptor beta interference effect, is beneficial to calculation of the toxicity equivalent, and has a good application prospect in the field of in-vitro biological test of the retinoic acid receptor beta interference effect. The problem that only the EC20 value or the EC50 value of the water environment sample can be calculated in the previous research is solved, and the method is beneficial to visual display of toxicity results and data use of an environment management department. The application also reveals that the pentachlorophenol toxicity concentration corresponding to the background value of the interference effect of the retinoic acid receptor beta of surface water in China is 6.90 [mu] g / L.

Description

Technical Field

[0001] The present application relates to the field of environmental detection technology, and in particular to an application of pentachlorophenol as a standard reference substance in detecting the interference effect of environmental samples on retinoic acid receptor β. Background Art

[0002] High-throughput in vitro biological testing of the comprehensive toxicity of water environment samples can directly obtain biological toxicity information under the coexistence of many pollutants in the water environment. By converting the comprehensive toxicity of water environment samples into the toxic equivalent concentration of a certain standard reference substance, the relative toxicity of water environment samples can be reflected. For example, the most widely used method is to convert the interference effect of environmental pollutants on aromatic hydrocarbon receptors into the concentration equivalent of dioxins. The conversion of the results of this in vitro comprehensive toxicity test into the toxicity of standard reference substances has become one of the important means of assessing and diagnosing water environment pollution.

[0003] The World Health Organization (WHO) points out that environmental endocrine disruptors refer to exogenous chemical substances or mixtures that can affect the function of the endocrine system of animals and cause damage to individuals, their offspring, or groups (partial subgroups). At present, thousands of environmental substances in the water environment have been identified as endocrine disruptors. In addition to interfering with the estrogen and androgen systems, many of these pollutants also affect the retinoic acid system through retinoic acid receptors, thereby affecting the growth and development of mammals, such as the differentiation and development of the brain, trunk, and limbs, and causing eye keratinization, dry eyes, and even blindness, increasing the risk of cancer.

[0004] At present, inhibitors of retinoic acid receptor β have been detected in environmental water bodies such as rivers. Considering that retinoic acid receptor β is most widely distributed throughout the body, environmental pollutants can interfere with multiple systems of the human body through retinoic acid receptor β. Therefore, the determination of the interference ability of retinoic acid receptor β in environmental pollutants can provide data support for the health risk assessment of environmental pollutants. However, there is a lack of standard reference substances for the inhibitory effect of retinoic acid receptor β in water environment samples at home and abroad, and there is no value of the effect trigger value of in vitro biological tests (there are estrogen, androgen and thyroid hormone, etc.). Therefore, it is urgent to screen standard reference compounds for in vitro biological tests of retinoic acid receptor β interference effects in water environment samples and determine the surface water effect trigger value based on ecological protection that conforms to my country's national conditions. Summary of the invention

[0005] Based on this, one or more embodiments of the present application provide an application of pentachlorophenol as a standard reference substance in detecting the interference effect of environmental samples on retinoic acid receptor β, and further determine that the equivalent concentration of pentachlorophenol of 10.77 μg / L is the surface water effect trigger value based on ecological protection of my country's water environment. The following technical solutions are included:

[0006] One or more embodiments of the present application provide a use of pentachlorophenol as a standard reference substance in detecting the interference effect of environmental samples on retinoic acid receptor β.

[0007] In some embodiments of the present application, the trigger value of the interference effect of the environmental sample on retinoic acid receptor β corresponds to a pentachlorophenol toxicity concentration of 10.77 μg / L.

[0008] In some embodiments of the present application, the background value of the interference effect of the environmental sample on retinoic acid receptor β corresponds to a pentachlorophenol toxicity concentration of 6.90 μg / L.

[0009] In some embodiments of the present application, the toxicity threshold value of the interference effect of the environmental sample on retinoic acid receptor β corresponds to a pentachlorophenol toxicity concentration of 1.62 μg / L.

[0010] In some embodiments of the present application, the water environment management value of pentachlorophenol is 9.0 μg / L.

[0011] In some embodiments of the present application, the environmental sample includes a water sample.

[0012] In some embodiments of the present application, the water source of the water sample includes surface water.

[0013] In some embodiments of the present application, the surface water includes source water.

[0014] In some embodiments of the present application, the surface water includes water from a common section.

[0015] In some embodiments of the present application, the detection comprises an in vitro biological test.

[0016] The details of one or more embodiments of the present application are set forth in the description which follows, and other features, objects, and advantages of the present application will be apparent from the description and its claims. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] In order to more clearly illustrate the technical solutions in the embodiments of the present application and to more completely understand the present application and its beneficial effects, the following is a brief introduction to the drawings required for the description of the embodiments. Obviously, the drawings described below are only some embodiments of the present application, and those skilled in the art can obtain other drawings based on these drawings without creative work.

[0018] Figure 1 To compare the interference types and intensities of the substances with high detection rates in my country's surface waters after testing the interference effects of retinoic acid receptor β.

[0019] Figure 2The dose-effect relationship curves of the interference effects of various substituted phenolic compounds on retinoic acid receptor β;

[0020] Figure 3 is the pentachlorophenol equivalent concentration of the RXRβ inhibitory effect of sewage in the Yangtze River Basin;

[0021] Figure 4 It is the pentachlorophenol equivalent concentration of RXRβ inhibitory effect in the Yangtze River Basin section and drinking water sources. DETAILED DESCRIPTION

[0022] The present application will be further described in detail below in conjunction with the accompanying drawings, embodiments and examples. It should be understood that these embodiments and examples are only used to illustrate the present application and are not used to limit the scope of the present application. The purpose of providing these embodiments and examples is to make the understanding of the disclosure of the present application more thorough and comprehensive. It should also be understood that the present application can be implemented in many different forms, is not limited to the embodiments and examples described herein, and those skilled in the art can make various changes or modifications without violating the connotation of the present application, and the equivalent form obtained also falls within the protection scope of the present application. In addition, in the description below, a large number of specific details are given in order to provide a more comprehensive understanding of the present application, and it should be understood that the present application can be implemented without one or more of these details.

[0023] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art to which this application belongs. The terms used herein in the specification of this application are only for the purpose of describing implementation modes and embodiments and are not intended to limit this application.

[0024] the term

[0025] Unless otherwise specified or incompatible herewith, the terms and phrases used herein shall have the following meanings:

[0026] The terms "and / or", "or / and", and "and / or" used in this article include any one of two or more related listed items, and also include any and all combinations of related listed items, and the arbitrary and all combinations include any two related listed items, any more related listed items, or a combination of all related listed items. It should be noted that when at least three items are connected by at least two conjunctions selected from "and / or", "or / and", and "and / or", it should be understood that in this application, the technical solution undoubtedly includes technical solutions that are all connected by "logical and", and undoubtedly includes technical solutions that are all connected by "logical or". For example, "A and / or B" includes three parallel solutions of A, B and A+B. For example, the technical solution of "A, and / or, B, and / or, C, and / or, D" includes any one of A, B, C, and D (that is, the technical solution that is all connected by "logical OR"), and also includes any and all combinations of A, B, C, and D, that is, the combination of any two or any three of A, B, C, and D, and also includes the combination of four of A, B, C, and D (that is, the technical solution that is all connected by "logical AND").

[0027] In the present application, "plurality", "multiple", "multiple times", "multiples", etc., unless otherwise specified, refer to a number greater than 2 or equal to 2. For example, "one or more" means one or greater than or equal to two.

[0028] As used herein, "combination thereof", "any combination thereof", "any combination thereof" etc. include all suitable combinations of any two or more of the listed items.

[0029] Herein, the “suitable” mentioned in “suitable combination”, “suitable method”, “any suitable method”, etc., shall be based on the ability to implement the technical solution of this application, solve the technical problems of this application, and achieve the expected technical effects of this application.

[0030] Herein, “preferred”, “better”, “more preferred” and “suitable” are merely used to describe implementation methods or examples with better effects, and it should be understood that they do not constitute limitations on the scope of protection of this application.

[0031] In the present application, "further", "further", "particularly" and the like are used for descriptive purposes to indicate differences in content, but should not be construed as limiting the scope of protection of the present application.

[0032] In this application, "optionally", "optional", and "optional" mean optional or dispensable, that is, any one of the two parallel schemes of "yes" or "no". If multiple "options" appear in a technical solution, unless otherwise specified and there is no contradiction or mutual restriction, each "optional" is independent.

[0033] In the present application, the terms "first", "second", "third", "fourth", etc. in "the first aspect", "the second aspect", "the third aspect", "the fourth aspect", etc. are used only for descriptive purposes and cannot be understood as indicating or implying relative importance or quantity, nor can they be understood as implicitly indicating the importance or quantity of the indicated technical features. Moreover, "first", "second", "third", "fourth", etc. only serve the purpose of non-exhaustive enumeration and description, and it should be understood that they do not constitute a closed limitation on quantity.

[0034] In the present application, the technical features described in an open manner include closed technical solutions composed of the listed features, and also include open technical solutions containing the listed features.

[0035] In the present application, when it comes to numerical intervals (i.e., numerical ranges), unless otherwise specified, the optional numerical distribution is considered continuous within the above numerical interval, and includes the two numerical endpoints (i.e., the minimum and maximum values) of the numerical range, and each numerical value between the two numerical endpoints. Unless otherwise specified, when the numerical interval only refers to the integers within the numerical interval, it includes the two endpoint integers of the numerical range, and each integer between the two endpoints. In this article, it is equivalent to directly listing each integer, such as t is an integer selected from 1 to 10, indicating that t is any integer selected from the group of integers consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10. In addition, when multiple ranges are provided to describe features or characteristics, these ranges can be combined. In other words, unless otherwise specified, the ranges disclosed herein should be understood to include any and all subranges included therein.

[0036] The temperature parameters in this application, unless otherwise specified, are allowed to be either constant temperature treatment or to vary within a certain temperature range. It should be understood that the constant temperature treatment allows the temperature to fluctuate within the accuracy range controlled by the instrument. Fluctuations within the range of ±5°C, ±4°C, ±3°C, ±2°C, and ±1°C are allowed.

[0037] In the present application, % (w / w) and wt% both represent weight percentage, % (v / v) refers to volume percentage, and % (w / v) refers to mass volume percentage.

[0038] Environmental samples refer to various substances collected from the environment for the purpose of analyzing and evaluating environmental quality. There are many types of environmental samples, mainly including the following categories: atmospheric environmental samples, water environmental samples, soil environmental samples, solid waste samples, biological samples, etc.

[0039] High-throughput in vitro biological testing of the comprehensive toxicity of water environment samples with standard reference substances can directly obtain biological toxicity information under the coexistence of many pollutants in the water environment. By converting the comprehensive toxicity of water environment samples into the toxic equivalent concentration of a certain standard reference substance, the relative toxicity of water environment samples can be reflected. For example, the most widely used method is to convert the interference effect of environmental pollutants on aromatic hydrocarbon receptors into the concentration equivalent of dioxins. The conversion of the results of this in vitro toxicity comprehensive test into the toxicity of standard reference substances has become one of the important means of assessing and diagnosing water environment pollution.

[0040] The 5% hazard concentration (HC5) is an important ecological risk assessment indicator, which refers to the concentration of a substance that can cause harm to 5% of species in a specific environment. This indicator is often used to assess the impact of environmental pollution on biodiversity, especially in the establishment of water quality benchmarks and aquatic toxicity studies. The estimation of HC5 involves statistical analysis of toxicity data of different species to determine the concentration of a substance that may cause harm to a certain proportion of biological populations in a specific environment.

[0041] The toxic equivalent level at a safe concentration is an important indicator of the impact of chemical substances on organisms. When evaluating the toxicity of chemical substances, the safe concentration is a key parameter, which refers to the maximum concentration of chemical substances allowed to exist without affecting the health of organisms. The toxic equivalent level is an indicator used to quantify this impact, which reflects the potential degree of harm of chemical substances to organisms.

[0042] Effect-Based Trigger Values ​​(EBT) refer to the critical value at which a certain pollutant or indicator in the environment may begin to have adverse effects on the ecosystem or human health when it reaches a certain concentration or level. It is determined based on a comprehensive assessment and research of various factors in the environment.

[0043] At present, there is a lack of standard reference substances commonly found in water environment samples that can be used as the inhibitory effect of retinoic acid hormone receptor β. Although the effect trigger values ​​of in vitro biological tests for the interference effects of hormone receptors such as estrogen, androgen and thyroid in water environment samples have been reported in the literature, there are no effect trigger values ​​of in vitro biological tests for the interference effects of all retinoic acid hormone receptors (α, β, γ). The effect trigger values ​​of foreign in vitro biological tests are mainly constructed based on the water quality conditions of the country or region, and there is currently no effect trigger value of retinoic acid hormone receptor β in the water environment that meets the water quality conditions in my country.

[0044] The present application screens out a standard reference compound for in vitro biological testing of the retinoic acid receptor β interference effect on aquatic samples. The compound has a high detection rate in river waters in my country, and the dose-effect relationship curve for the retinoic acid receptor β interference effect has a good linear range, which is conducive to the calculation of toxic equivalents and solves the problem that only the EC20 or EC50 values ​​of aquatic samples could be calculated in previous studies. It is conducive to the intuitive display of toxicity results and the use of data by environmental management departments.

[0045] This application determines the effect trigger value of the water environment of the retinoic acid hormone receptor β inhibitory effect that meets the surface water quality conditions in my country. This effect trigger value needs to comprehensively consider the background value of pollutants in my country, the starting point of the toxic effect of retinoic acid hormone receptor β, and the protection level of native water species in my country. The effect trigger value determined in this application can provide data for the environmental monitoring department to monitor endocrine disruptors; and provide support for the safety evaluation of chemicals and the environmental management of new chemical substances for the environment and related departments.

[0046] The present application embodiment provides an application of pentachlorophenol as a standard reference substance in detecting the interference effect of environmental samples on retinoic acid receptor β.

[0047] In some of the examples, the trigger value of the interference effect of the environmental sample on retinoic acid receptor β corresponds to a toxic concentration of pentachlorophenol of 10.77 μg / L.

[0048] In some of the examples, the background value of the interference effect of the environmental samples on retinoic acid receptor β corresponds to a pentachlorophenol toxicity concentration of 6.90 μg / L.

[0049] In some of the examples, the toxicity threshold value of the interference effect of the environmental samples on retinoic acid receptor β corresponds to a pentachlorophenol toxicity concentration of 1.62 μg / L.

[0050] In some of the examples, the water environment management value of pentachlorophenol is 9.0 μg / L.

[0051] In some examples, the environmental sample includes a water sample.

[0052] In some examples, the source of the water sample includes surface water.

[0053] In some of these examples, the surface water includes source water.

[0054] In some of these examples, the surface water includes water from a common cross section.

[0055] In some of these examples, the testing includes in vitro biological testing.

[0056] The embodiments of the present application will be described in detail below in conjunction with examples. It should be understood that these examples are only used to illustrate the present application and are not intended to limit the scope of the present application. The experimental methods for which specific conditions are not specified in the following examples are preferably referred to the guidance provided in the present application, and can also be based on the experimental manual or normal conditions in this area, can also be based on the conditions recommended by the manufacturer, or refer to experimental methods known in the art.

[0057] In the following specific embodiments, the measured parameters of raw material components may have slight deviations within the range of weighing accuracy unless otherwise specified. For temperature and time parameters, acceptable deviations caused by instrument test accuracy or operation accuracy are allowed.

[0058] 1. Reference material screening

[0059] The retinoic acid receptor β interference effect was detected on highly detected substances in my country's surface water bodies, and the retinoic acid receptor β interference effect was determined on various common pollutants in water bodies using a two-hybrid yeast experiment with recombinant retinoic acid receptor β. Among them, pollutants with strong interference effects on retinoic acid receptor β and with high concentrations in my country's surface water were screened out, and substituted phenols were screened out as highly active interfering substances.

[0060] Then, the substituted phenolic compounds with high detection rate in my country's surface water bodies were tested for the above-mentioned interference effect of retinoic acid receptor β. By obtaining the complete dose-effect relationship curves of these compounds, substituted phenols with higher activity, good dose-effect relationship curves and longer linear intervals can be screened as standard reference substances. By comparing the dose-effect relationship curves between various substituted phenols, pentachlorophenol was finally selected as the standard reference substance for the inhibitory effect of retinoic acid receptor β.

[0061] according to Figure 1 It can be seen that substituted phenolic compounds generally have a stronger interference effect on retinoic acid receptor β.

[0062] according to Figure 2 It can be seen that pentachlorophenol has higher activity, better dose-effect relationship curve and longer linear range than other substituted phenolic compounds, and is suitable as a standard reference substance for the inhibitory effect of retinoic acid receptor β.

[0063] Among them, the method for determining the interference effect of retinoic acid receptor β is as follows:

[0064] (1) 1 mL of freshly frozen and thawed recombinant retinoic acid receptor β yeast strain (recombinant retinoic acid receptor β gene double hybrid Saccharomyces cerevisiae) was centrifuged and inoculated into 20 mL of SD culture medium, and cultured overnight in an air bath shaker at 150 rpm and 37°C. After diluting the bacterial solution 10 times, the absorbance was measured at 600 nm and the bacterial solution was diluted to make the OD600 value 0.75. References for yeast strain construction: Li Jian et al., Study on the interference effect of retinoic acid X receptor in sewage treatment plant effluent, Journal of Beijing Institute of Technology, 2011, 31(02).

[0065] (2) Take 990 μL of bacterial solution into an EP tube, add 5 μL of the endogenous retinoic acid receptor β inducer (9-cis retinoic acid) and 5 μL of the sample to be tested (various common pollutants) and mix well; transfer 200 μL of each of the above solutions to a 96-well plate in turn. After shaking and culturing at 800 rpm and 37°C for 2 hours, measure the OD600.

[0066] (3) Discard 150 μL of the bacterial solution in the culture plate and add 120 μL of test buffer (basic buffer solution: dissolve 21.51 g of Na2HPO4·12H2O, 6.22 g of NaH2PO4·2H2O, 0.25 g of MgSO4·7H2O and 0.75 g of KCl and make up to 1 L of distilled water, sterilize after full dissolution, and store in a refrigerator at 4°C for later use; test buffer solution: add 3.33 ml of 0.1% SDS solution and 270 uL of β-mercaptoethanol to every 100 mL of basic buffer), then add 20 μL of chloroform to break the cell wall, shake on a shaker at 1200 rpm for 10 min; add 40 μL of ONPG to start the enzyme reaction, place on a shaker at 30°C, and start timing the reaction time at 800 rpm until a distinct yellow color appears (within 2 h). Add 100 μL Na2CO3 solution to terminate the reaction; shake and mix on a constant temperature plate shaker at 400 rpm for 10 min. Transfer the supernatant to an ELISA plate and measure the OD420 value at a wavelength of 420 nm.

[0067] (4) Based on the OD420 and OD600 values, the enzyme activity values ​​of the test sample, the inducer of endogenous retinoic acid receptor β (9-cis retinoic acid) and the solvent control (DMSO) as well as the inhibition rate of the test sample relative to the positive control are calculated, and the dose-effect relationship curve of the test sample and the equivalent concentration of pentachlorophenol are obtained.

[0068] 2. Calculation of the effect trigger value for the inhibitory effect of retinoic acid hormone receptor β

[0069] Based on the toxic equivalent level (background-BEQ) of the background investigation, the toxic equivalent level at 5% hazard concentration (HC5-BEQ) and the toxic equivalent level at the safe concentration, that is, the minimum effect concentration (safe-BEQ), the trigger value for the inhibitory effect of retinoic acid hormone receptor β is calculated together.

[0070] 2.1 By looking up the environmental standards of pentachlorophenol in my country's surface water standards and the values ​​of HC5 (5% hazardous concentration) of pentachlorophenol in my country's surface water bodies in the literature, the water environmental management value of pentachlorophenol for my country's surface water quality is 9.0μg / L (the target standard limit of the specific project of the centralized drinking water surface water source), and the HC5 value based on native species in my country's surface water is 10.77μg / L. Zhang Yahui, Cao Ying, Li Ji, et al. Ecological risk assessment of three chlorophenol mixtures in my country's main rivers [J]. Environmental Science Research, 2012, 25(4): 385-390.

[0071] 2.2 Water samples were collected in the Yangtze River Basin and the Yellow River Basin. The water samples collected included water sources, sections and water bodies near sewage outlets, with a total of 48 sampling points. These points are located in 12 cities from the upper reaches of the Yangtze River to the lower reaches of the Yangtze River, namely Luzhou, Chongqing, Wanzhou, Yichang, Yueyang, Wuhan, Jiujiang, Tongling, Nanjing, Taizhou, Nantong, and Shanghai. The collected water samples were stored in brown glass bottles that had been washed with chromic acid and rinsed with water samples in advance. The samples were immediately transported back to the laboratory and enriched within 24 hours. The water samples were filtered through a glass cellulose membrane (APFF, MILLIPORE, USA) and then enriched with an HLB column (500 mg, OASIS, USA), and then gradient eluted with solvents such as dichloromethane and methanol. The eluate was concentrated by rotary evaporation, dehydrated by adding anhydrous sodium sulfate, and the replacement solvent was DMSO (99.5% GC, SIGMA), fixed to 0.2 mL, and stored at -20°C for the determination of the interference effect of retinoic acid receptor β. The determination method refers to the determination method of the interference effect of retinoic acid receptor β recorded under Item 1. The measured toxicity data was regressed through the standard curve of the inhibitory effect of pentachlorophenol on retinoic acid hormone receptor β, and the pentachlorophenol equivalent concentration of the interference effect of retinoic acid receptor β in water samples of the Yangtze River Basin and the Yellow River Basin in my country was obtained.

[0072] The background value is calculated by summing up the equivalent concentration of pentachlorophenol at each sampling point and then calculating the average value. The background value of the retinoic acid receptor β inhibitory effect in major water bodies in my country is 6.90 μg / L.

[0073] 2.3 Calculate the toxicity threshold value of the inhibitory effect of pentachlorophenol on retinoic acid receptor β, that is, the safe-BEQ value. This value is obtained based on the dose-effect relationship curve of the inhibitory effect of pentachlorophenol on retinoic acid receptor β, that is, by Figure 1The dose-effect relationship curve of pentachlorophenol in water was used to obtain its toxicity starting value, i.e., the EC10 value was 6.1×10 -9 mol / L, EC10 = (10% effect value - minimum effect value) / (maximum effect value - minimum effect value) × concentration range + minimum concentration value, combined with the molecular weight of pentachlorophenol is 266.3 g / mol, the concentration of safe-BEQD is 1.62 μg / L.

[0074] 2.4 The in vitro biological toxicity test effect trigger value of the retinoic acid receptor β inhibitory effect in my country's water quality was determined by comprehensively calculating the background value of the retinoic acid receptor β inhibitory effect of major water bodies in my country (6.90 μg / L), the toxicity threshold value of the retinoic acid receptor β inhibitory effect of pentachlorophenol (safe-BEQ value, 1.62 μg / L), and the water environment management value of pentachlorophenol in my country's water quality (9.0 μg / L). The rules adopted are as follows:

[0075] (1) If background-BEQ is less than HC5-BEQ, the effect trigger value is HC5-BEQ;

[0076] (2) If background-BEQ is less than HC5-BEQ by more than 100 times, the effect trigger value is (2-10)×safe-BEQ;

[0077] (3) If the background-BEQ and HC5-BEQ data are close, the effect trigger value is the HC5 95% confidence interval;

[0078] (4) If background-BEQ is greater than HC5-BEQ data, the effect trigger value is (2-5)×background-BEQ.

[0079] According to the actual test results of my country's surface water bodies in this application, the background-BEQ was 6.90μg / L, while the HC5-BEQ was 10.77μg / L, which belongs to the first case. Therefore, it can be inferred that the effect trigger value is 10.77μg / L.

[0080] 3. In vitro biological test of the interference effect of retinoic acid receptor β in vitro

[0081] The sampling points in the Yangtze River Basin include 12 cities from the upper reaches of the Yangtze River to the lower reaches of the Yangtze River, and 48 sampling points (located in 12 typical cities including Luzhou, Chongqing, Wanzhou, Yichang, Yueyang, Wuhan, Jiujiang, Tongling, Nanjing, Taizhou, Nantong, and Shanghai). Figure 3 P1 to P12 are the points close to the sewage outlets in the Yangtze River in these 12 cities. Figure 4D1-1 to D12-2 are 24 points of ordinary sections of the Yangtze River in these 12 cities, and S1 to S12 are points where the Yangtze River serves as water sources in these 12 cities.

[0082] Figure 3 , Figure 4 Characterization data: It proves that the water samples of my country's surface water bodies, such as the water samples near the sewage outlets of the Yangtze River, as water sources and ordinary sections, are all below the effect trigger value of the retinoic acid receptor β inhibitory effect and are relatively safe.

[0083] The technical features of the above-mentioned implementation modes and examples can be combined in any appropriate manner. To make the description concise, not all possible combinations of the technical features in the above-mentioned implementation modes and examples are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0084] The above-described embodiments only express several implementation methods of the present application, which is convenient for understanding the technical solution of the present application in detail, but it cannot be understood as a limitation on the scope of protection of the patent application. It should be pointed out that for ordinary technicians in this field, without departing from the concept of the present application, several deformations and improvements can be made, which all belong to the protection scope of the present application. In addition, it should be understood that after reading the above-mentioned teaching content of the present application, the technicians in this field can make various changes or modifications to the present application, and the equivalent forms obtained also fall within the protection scope of the present application. It should also be understood that the technical solutions obtained by the technicians in this field through logical analysis, reasoning or limited experiments on the basis of the technical solutions provided in the present application are all within the protection scope of the claims attached to the present application. Therefore, the protection scope of the patent of the present application shall be based on the content of the attached claims, and the description and drawings can be used to explain the content of the claims.

Claims

1. Application of pentachlorophenol as a standard reference substance in detecting the interference effect of environmental samples on retinoic acid receptor β.

2. The use according to claim 1, characterized in that: The trigger value of the interference effect of the environmental sample on retinoic acid receptor β corresponds to a pentachlorophenol toxicity concentration of 10.77 μg / L.

3. The use according to claim 2, characterized in that: The background value of the interference effect of the environmental sample on retinoic acid receptor β corresponds to a pentachlorophenol toxicity concentration of 6.90 μg / L.

4. The use according to claim 2, characterized in that: The toxicity starting value of the interference effect of the environmental sample on retinoic acid receptor β corresponds to a pentachlorophenol toxicity concentration of 1.62 μg / L.

5. The use according to claim 2, characterized in that: The water environment management value of pentachlorophenol is 9.0 μg / L.

6. The use according to any one of claims 1 to 5, characterized in that The environmental sample includes a water sample.

7. The use according to claim 6, characterized in that: The water source of the water sample includes surface water.

8. The use according to claim 7, characterized in that: The surface water includes source water.

9. The use according to claim 8, characterized in that: The surface water includes water in ordinary sections.

10. The use according to any one of claims 1 to 5 and 7 to 9, characterized in that Testing includes in vitro biological tests.

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