CN114062074A - Method for discriminating toxicity of organic composite polluted sediment - Google Patents
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- 239000013049 sediment Substances 0.000 title claims abstract description 76
- 231100000419 toxicity Toxicity 0.000 title claims abstract description 37
- 230000001988 toxicity Effects 0.000 title claims abstract description 37
- 238000000034 method Methods 0.000 title claims abstract description 36
- 239000002131 composite material Substances 0.000 title claims abstract description 32
- 238000012360 testing method Methods 0.000 claims abstract description 54
- 231100000331 toxic Toxicity 0.000 claims abstract description 24
- 238000000338 in vitro Methods 0.000 claims abstract description 19
- 230000002588 toxic effect Effects 0.000 claims abstract description 19
- 238000012216 screening Methods 0.000 claims abstract description 17
- 231100000820 toxicity test Toxicity 0.000 claims abstract description 17
- 238000004458 analytical method Methods 0.000 claims abstract description 14
- 238000001228 spectrum Methods 0.000 claims abstract description 10
- 231100001234 toxic pollutant Toxicity 0.000 claims abstract description 9
- 241000256135 Chironomus thummi Species 0.000 claims abstract description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 48
- 241000256128 Chironomus <genus> Species 0.000 claims description 29
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 24
- 239000011347 resin Substances 0.000 claims description 24
- 229920005989 resin Polymers 0.000 claims description 24
- 239000000284 extract Substances 0.000 claims description 20
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 16
- 238000003756 stirring Methods 0.000 claims description 12
- 230000003203 everyday effect Effects 0.000 claims description 9
- 239000007788 liquid Substances 0.000 claims description 9
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 claims description 9
- 238000004587 chromatography analysis Methods 0.000 claims description 6
- 239000008367 deionised water Substances 0.000 claims description 6
- 229910021641 deionized water Inorganic materials 0.000 claims description 6
- 238000000605 extraction Methods 0.000 claims description 6
- 239000011521 glass Substances 0.000 claims description 6
- 238000004366 reverse phase liquid chromatography Methods 0.000 claims description 6
- 239000004576 sand Substances 0.000 claims description 6
- 238000005406 washing Methods 0.000 claims description 6
- 238000005286 illumination Methods 0.000 claims description 3
- 230000002452 interceptive effect Effects 0.000 claims description 3
- 150000002894 organic compounds Chemical class 0.000 claims description 3
- 230000003044 adaptive effect Effects 0.000 claims 1
- 238000010998 test method Methods 0.000 claims 1
- 239000002957 persistent organic pollutant Substances 0.000 abstract description 8
- 238000001514 detection method Methods 0.000 abstract description 2
- 239000000243 solution Substances 0.000 description 9
- 239000003344 environmental pollutant Substances 0.000 description 8
- 231100000719 pollutant Toxicity 0.000 description 8
- ONKNPOPIGWHAQC-UHFFFAOYSA-N galaxolide Chemical compound C1OCC(C)C2=C1C=C1C(C)(C)C(C)C(C)(C)C1=C2 ONKNPOPIGWHAQC-UHFFFAOYSA-N 0.000 description 6
- 238000011097 chromatography purification Methods 0.000 description 3
- 239000011259 mixed solution Substances 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 231100000041 toxicology testing Toxicity 0.000 description 3
- DNRJTBAOUJJKDY-UHFFFAOYSA-N 2-Acetyl-3,5,5,6,8,8-hexamethyl-5,6,7,8- tetrahydronaphthalene Chemical compound CC(=O)C1=C(C)C=C2C(C)(C)C(C)CC(C)(C)C2=C1 DNRJTBAOUJJKDY-UHFFFAOYSA-N 0.000 description 2
- 241000251468 Actinopterygii Species 0.000 description 2
- 241000402754 Erythranthe moschata Species 0.000 description 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 239000001301 oxygen Substances 0.000 description 2
- 229910052760 oxygen Inorganic materials 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 1
- 238000003723 Smelting Methods 0.000 description 1
- XKMRRTOUMJRJIA-UHFFFAOYSA-N ammonia nh3 Chemical compound N.N XKMRRTOUMJRJIA-UHFFFAOYSA-N 0.000 description 1
- 238000003889 chemical engineering Methods 0.000 description 1
- 239000000356 contaminant Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000006866 deterioration Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 229910001385 heavy metal Inorganic materials 0.000 description 1
- 239000000543 intermediate Substances 0.000 description 1
- 239000000575 pesticide Substances 0.000 description 1
- 238000005067 remediation Methods 0.000 description 1
- 239000010865 sewage Substances 0.000 description 1
- 231100000048 toxicity data Toxicity 0.000 description 1
- 238000003911 water pollution Methods 0.000 description 1
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
- G01N33/5008—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
- G01N33/5014—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 toxicity
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Abstract
The invention discloses a method for screening toxicity of organic composite polluted sediments, which comprises the following steps: 1) domesticating chironomid larvae; 2) pretreating organic composite pollution sediments; 3) extracting sediment extracting solution; 4) and (3) testing: separating interference components of the sediment extracting solution to obtain components for biological toxicity test, finding out toxic components by combining with the midge larva in-vitro toxicity test, then carrying out scanning analysis and spectrum library comparison on the toxic components, and tracking specific toxic pollutant. The method for screening the toxicity of the organic composite polluted sediments adopts common biological tests, can economically and visually reflect the toxicity of the sediments, meets the test requirements of most laboratories, can be popularized and used, can narrow the detection range, defines specific toxic organic pollutants, and has low cost in the whole process.
Description
Technical Field
The invention relates to a method for screening toxicity of organic composite polluted sediments.
Background
Since the 20 th century, with the rapid development of industries such as smelting, chemical engineering, pesticides and medicines, a large amount of industrial, agricultural and domestic sewage is discharged into water, so that the water is seriously polluted. After entering the water body, the pollutants are gradually enriched in the water body sediments, and the pollutants in the sediments are released into the water body again through the chemical and physical process to become a secondary pollution source, so that the water quality and the ecological system are influenced. Thus, sediment is a "source" and "sink" of water pollution. Furthermore, even if the source of pollution ceases to contaminate the water, the contaminated sediment may cause the water quality to deteriorate. High concentrations of contaminants accumulated in the sediment can be harmful to benthic organisms. For every body of water, contaminated sediments pose a potential risk to fish and humans, as well as to wild animals eating fish. Contaminated sediments also cause ecological and economic losses to aquatic resources, such as deterioration of habitats and expensive remediation costs.
In recent years, a large amount of pollutants including three types of organic pollutants, heavy metals and ammonia nitrogen are detected in sediments. Organic pollution is more serious. The toxicity of organic pollutants in water sediments is often the result of the combined action of multiple pollutants and various complex intermediates thereof, and the complexity and complexity of the pollution make the traditional characteristic pollutant-based toxicity evaluation method inapplicable. The extensive extension of the chemical analysis object, even the full scan of known organic pollutants, may partially compensate this drawback, however, the large amount of time, financial consumption and the lack of toxicity data limit the further application of the method. Therefore, in order to accurately evaluate and control the pollution of the water body sediments, on one hand, the toxicity of the water body sediments needs to be known, and on the other hand, the types of direct or indirect pollutants which generate toxic effects or specific pollutants need to be known.
When the toxicity of organic pollutants in the sediment is screened, some unknown pollutants cannot be tracked due to the coexistence of multiple organic pollutants. Therefore, it is an urgent need to solve the problem of determining the current biological toxicity of organic pollutants in sediments, tracing toxic pollutants, and finally identifying toxic pollutants.
Disclosure of Invention
In view of the above, the invention aims to provide a method for screening the toxicity of organic composite polluted sediments.
The technical scheme provided by the invention is as follows: a method for screening toxicity of organic composite polluted sediments comprises the following steps:
1) domesticating chironomid larvae
Collecting chironomus larvae as a test organism, and then domesticating the chironomus larvae for not less than 7 days;
2) pretreatment of organic composite contaminated deposits
Storing the organic composite pollution sediments to be treated in a refrigerator, and uniformly stirring;
3) extraction of sediment extract
Taking 200-500 g of wet sediment, 200-500 ml of medium-hardness water and 10-20g of XAD resin, stirring for 24 hours at 23 ℃ in a dark place, then recovering the resin, washing with deionized water, and then respectively extracting with mixed liquid of acetone, n-hexane and acetone to obtain extract liquid, namely: a sediment extract;
4) testing
Separating interference components of the sediment extracting solution to obtain components for biological toxicity test, finding out toxic components by combining with the midge larva in-vitro toxicity test, then carrying out scanning analysis and spectrum library comparison on the toxic components, and tracking specific toxic pollutant.
Preferably, in the step 1), feeding 1-2 times every day and changing water every 3 days in the process of domesticating the chironomid larvae.
Further preferably, in step 4), the interfering components of the sediment extract are separated by normal phase chromatography and reverse phase chromatography.
Further preferably, in the step 4), the method for testing the toxicity of the chironomus larvas in vitro is as follows:
first, 12-well plates were taken for testing, and then the components for biotoxicity testing were dissolved in dimethyl sulfoxide and poured into the wells of 12-well plates, respectively, and then 5 chironomid larvae, 4-5 ml of medium hardness water and a small amount of clean sand were added per well for testing.
Further preferably, the test time of the toxicity of the chironomus larvas in vitro is 72 hours, the test temperature is set to be 23-25 ℃, the illumination is 16-18 hours per day, the darkness is 8-6 hours, and the whole process is not changed with water and is not fed.
Further preferably, a glass lining adapted to the hole of the 12-hole plate is arranged in the hole of the 12-hole plate to prevent organic compounds from being adsorbed on the hole plate.
The method for screening the toxicity of the organic composite polluted sediment provided by the invention adopts common biological tests, can economically and visually reflect the toxicity of the sediment, meets the test requirements of most laboratories, can be popularized and used, can narrow the detection range, defines specific toxic organic pollutants, and has low cost in the whole process.
Detailed Description
The invention will be further explained with reference to specific embodiments, without limiting the invention.
In order to solve the problem that the conventional method for screening the toxicity of the organic composite polluted sediment is complex, the invention provides a method for screening the toxicity of the organic composite polluted sediment, which comprises the following steps:
1) domesticating chironomid larvae
Collecting chironomus larvae as a test organism, and then domesticating the chironomus larvae for not less than 7 days;
2) pretreatment of organic composite contaminated deposits
Storing the organic composite pollution sediments to be treated in a refrigerator, and uniformly stirring;
3) extraction of sediment extract
Taking 200 g-500 g of wet sediment, 200-500 ml of medium-hardness water and 10-20g of XAD resin, stirring for 24 h at 23 ℃ in a dark place, then recovering the resin, washing with deionized water, and then respectively extracting with mixed liquid of acetone, n-hexane and acetone to obtain extract liquid, namely: a sediment extract;
4) testing
Separating interference components of the sediment extracting solution to obtain components for biological toxicity test, finding out toxic components by combining with the midge larva in-vitro toxicity test, then carrying out scanning analysis and spectrum library comparison on the toxic components, and tracking specific toxic pollutant.
As an improvement of the technical scheme, in the step 1), feeding 1-2 times every day and changing water every 3 days in the process of domesticating the chironomid larvae.
As an improvement of the technical scheme, in the step 4), the interfering components of the sediment extracting solution are separated through normal phase chromatography and reverse phase chromatography.
As an improvement of the technical scheme, in the step 4), the method for testing the toxicity of the chironomus larvas in vitro comprises the following steps:
first, 12-well plates were taken for testing, and then the components for biotoxicity testing were dissolved in dimethyl sulfoxide and poured into the wells of 12-well plates, respectively, and then 5 chironomid larvae, 4-5 ml of medium hardness water and a small amount of clean sand were added per well for testing.
As an improvement of the technical scheme, the test time of the toxicity of the chironomus larvas in vitro is 72 hours, the test temperature is set to be 23-25 ℃, the light is 16-18 hours per day, the darkness is 8-6 hours, and the whole process does not need to change water or feed.
As an improvement of the technical scheme, a glass lining adapted to the hole of the 12-hole plate is arranged in the hole of the 12-hole plate so as to prevent organic compounds from being adsorbed on the hole plate.
Example 1
The method for screening the toxicity of the organic composite polluted sediments comprises the following steps:
1) domesticating chironomid larvae
Taking Chironomus larvae collected in Liaohe river as test organisms, then domesticating the test organisms for 7 days, feeding the test organisms for 1 time every day, changing water every 3 days, wherein the water temperature is 25 ℃, and the pH value is 8 so as to ensure the optimal domesticating environment;
2) pretreatment of organic composite contaminated deposits
Storing the organic composite pollution sediments to be treated, which are taken from the area A, in a refrigerator, and uniformly stirring;
3) extraction of sediment extract
Taking 200g of wet sediment, 200ml of medium-hardness water and 10g of XAD resin, putting the wet sediment, 200ml of medium-hardness water and 10g of XAD resin into a 500ml conical flask, stirring the mixture for 24 hours at the temperature of 23 ℃ in a dark place at the speed of 500 revolutions per minute, then recovering the resin, washing the resin with deionized water for 3 times, and then respectively extracting the resin once with acetone and extracting the resin for 2 times with a mixed solution of normal hexane and acetone (the volume ratio is 1: 1) to obtain extract liquid, namely: a sediment extract;
4) testing
Separating interference components of the sediment extracting solution through a normal phase chromatography and a reverse phase chromatography to obtain components for biological toxicity test, finding out toxic components by combining with the midge larva in-vitro toxicity test, then, carrying out scanning analysis and spectrum library comparison on the toxic components, and tracking specific toxic pollutant;
the method for testing the in vitro toxicity of the chironomus larvas comprises the following steps:
firstly, taking a 12-hole plate for testing, wherein a glass lining adapted to the 12-hole plate is arranged in the 12-hole plate, then dissolving components for biological toxicity testing in dimethyl sulfoxide and pouring the components into the 12-hole plate, then adding 5 chironomus larvae, 4 ml of medium-hardness water and a small amount of clean sand into each hole to serve as attachments of organisms, testing, wherein the testing time is 72 hours, the testing temperature is set to be 23 ℃, the lighting is 16 hours every day, the darkness is 8 hours, and the whole process is not changed with water and is not fed.
5) Analysis of results
The sediment extracting solution is subjected to normal phase and reverse phase chromatographic purification combined with chironomus larvas in-vitro toxicity test, the toxicity effect of specific components is determined by calculating the death rate of chironomus larvas, then scanning analysis and spectrum library comparison are carried out on the toxicity components, the similarity of galaxolide is more than 80%, the fact that galaxolide has obvious toxicity effect on the chironomus larvas is proved, and LC50=7.5 ng/ml.
Example 2
The method for screening the toxicity of the organic composite polluted sediments comprises the following steps:
1) domesticating chironomid larvae
Chironomus larvae collected in Liaohe river are taken as test organisms, and then domesticated for 10 days, the test organisms are fed for 2 times every day, water is changed every 3 days, the water temperature is controlled at 23 ℃, the proper pH value is 7, and the dissolved oxygen is controlled to be more than 2 mg/L. Culturing with a light-to-dark ratio of 16: 8;
2) pretreatment of organic composite contaminated deposits
Storing the organic composite pollution sediments to be treated, which are taken from the area B, in a refrigerator, and uniformly stirring;
3) extraction of sediment extract
Taking 500g of wet sediment, 500ml of medium-hardness water and 20g of XAD resin, putting the wet sediment, 500ml of medium-hardness water and 20g of XAD resin into a 1000ml conical flask, stirring the mixture for 24 hours at the temperature of 23 ℃ in a dark place at the speed of 800 revolutions per minute, then recovering the resin, washing the resin with deionized water for 3 times, and then respectively extracting the resin once with acetone and extracting the resin for 3 times with a mixed solution of normal hexane and acetone (the volume ratio is 1: 1) to obtain extract liquid, namely: a sediment extract;
4) testing
Separating interference components of the sediment extracting solution through a normal phase chromatography and a reverse phase chromatography to obtain components for biological toxicity test, finding out toxic components by combining with the midge larva in-vitro toxicity test, then, carrying out scanning analysis and spectrum library comparison on the toxic components, and tracking specific toxic pollutant;
the method for testing the in vitro toxicity of the chironomus larvas comprises the following steps:
firstly, taking a 12-hole plate for testing, wherein a glass lining adapted to the 12-hole plate is arranged in the 12-hole plate, then dissolving components for biological toxicity testing in dimethyl sulfoxide and pouring the dimethyl sulfoxide into the 12-hole plate holes respectively, then adding 5 chironomus larvae, 5ml of medium-hardness water and a small amount of clean sand into each hole to serve as attachments of organisms, testing, wherein the testing time is 72 hours, the testing temperature is set to be 25 ℃, the illumination is 16 hours every day, the darkness is 8 hours, and the whole process is not changed with water or fed.
5) Analysis of results
The sediment extracting solution is subjected to normal phase and reverse phase chromatographic purification combined with chironomid larva in-vitro toxicity test, the toxicity effect of specific components is determined by calculating the death rate of chironomid larvae, then, scanning analysis and spectrum library comparison are carried out on toxic components, and the similarity of tonalid musk and galaxolide musk is more than 80%. Galaxolide and tonalid demonstrated significant toxic effects on chironomid larvae with LC50=9 ng/ml and 8.5 ng/ml.
Example 3
The method for screening the toxicity of the organic composite polluted sediments comprises the following steps:
1) domesticating chironomid larvae
Chironomus larvas collected in Liaohe river are used as test organisms and identified as local brook Chironomus, then the test organisms are domesticated for 7 days, the test organisms are fed for 1 time every day, water is changed every 3 days, the water temperature is controlled at 24 ℃, the proper pH value is 7, and the dissolved oxygen is controlled to be more than 2 mg/L. Culturing with a light-to-dark ratio of 18: 6;
2) pretreatment of organic composite contaminated deposits
Storing the organic composite pollution sediments to be treated from the region C in a refrigerator, and uniformly stirring;
3) extraction of sediment extract
Taking 300g of wet sediment, 300ml of medium-hardness water and 15g of XAD resin, putting the wet sediment, 300ml of medium-hardness water and 15g of XAD resin into a 1000ml conical flask, stirring the mixture for 24 hours at the temperature of 23 ℃ in a dark place at the speed of 700 revolutions per minute, then recovering the resin, washing the resin with deionized water for 3 times, and then respectively extracting the resin once with acetone and extracting the resin for 3 times with a mixed solution of normal hexane and acetone (the volume ratio is 1: 1) to obtain extract liquid, namely: a sediment extract;
4) testing
Separating interference components of the sediment extracting solution through a normal phase chromatography and a reverse phase chromatography to obtain components for biological toxicity test, finding out toxic components by combining with the midge larva in-vitro toxicity test, then, carrying out scanning analysis and spectrum library comparison on the toxic components, and tracking specific toxic pollutant;
the method for testing the in vitro toxicity of the chironomus larvas comprises the following steps:
firstly, taking a 12-hole plate for testing, wherein a glass lining adapted to the 12-hole plate is arranged in the 12-hole plate, then dissolving components for biological toxicity testing in dimethyl sulfoxide and pouring the dimethyl sulfoxide into the 12-hole plate holes respectively, then adding 5 chironomus larvae, 4 ml of medium-hardness water and a small amount of clean sand into each hole to serve as attachments of organisms, testing, wherein the testing time is 72 hours, the testing temperature is set to be 25 ℃, the light is set for 18 hours every day, the dark is set for 6 hours, and the whole process is not changed with water and fed.
5) Analysis of results
The sediment extracting solution is subjected to normal phase and reverse phase chromatographic purification combined with chironomus larvas in-vitro toxicity test, the toxicity effect of specific components is determined by calculating the death rate of chironomus larvas, then scanning analysis and spectrum library comparison are carried out on the toxicity components, the similarity of galaxolide is more than 80%, the fact that galaxolide has obvious toxicity effect on the chironomus larvas is proved, and LC50=8 ng/ml.
Claims (6)
1. A method for screening toxicity of organic composite polluted sediments is characterized by comprising the following steps:
1) domesticating chironomid larvae
Collecting chironomus larvae as a test organism, and then domesticating the chironomus larvae for not less than 7 days;
2) pretreatment of organic composite contaminated deposits
Storing the organic composite pollution sediments to be treated in a refrigerator, and uniformly stirring;
3) extraction of sediment extract
Taking 200-500 g of wet sediment, 200-500 ml of medium-hardness water and 10-20g of XAD resin, stirring for 24 hours at 23 ℃ in a dark place, then recovering the resin, washing with deionized water, and then respectively extracting with mixed liquid of acetone, n-hexane and acetone to obtain extract liquid, namely: a sediment extract;
4) testing
Separating interference components of the sediment extracting solution to obtain components for biological toxicity test, finding out toxic components by combining with the midge larva in-vitro toxicity test, then carrying out scanning analysis and spectrum library comparison on the toxic components, and tracking specific toxic pollutant.
2. The method for screening the toxicity of the organic composite polluted sediments according to claim 1, which is characterized by comprising the following steps: in the step 1), feeding 1-2 times every day and changing water every 3 days in the process of domesticating chironomus larvae.
3. The method for screening the toxicity of the organic composite polluted sediments according to claim 1, which is characterized by comprising the following steps: in step 4), the interfering components of the sediment extract are separated by normal phase chromatography and reverse phase chromatography.
4. The method for screening the toxicity of the organic composite polluted sediments according to claim 1, which is characterized by comprising the following steps: in the step 4), the test method for the in vitro toxicity of the chironomus larvas comprises the following steps:
first, 12-well plates were taken for testing, and then the components for biotoxicity testing were dissolved in dimethyl sulfoxide and poured into the wells of 12-well plates, respectively, and then 5 chironomid larvae, 4-5 ml of medium hardness water and a small amount of clean sand were added per well for testing.
5. The method for screening the toxicity of the organic composite polluted sediments according to claim 4, which is characterized by comprising the following steps: the test time of the toxicity of chironomus larvas in vitro is 72 hours, the test temperature is set to be 23-25 ℃, the illumination is 16-18 hours per day, the darkness is 8-6 hours, and the whole process does not change water or feed.
6. The method for screening the toxicity of the organic composite polluted sediments according to claim 4, which is characterized by comprising the following steps: and a glass lining adaptive to the hole of the 12-hole plate is arranged in the hole of the 12-hole plate so as to prevent organic compounds from being adsorbed on the hole plate.
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