CN111103386B - Method for evaluating full-biodegradable material ecotoxicity by using plant seeds - Google Patents
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Abstract
The invention discloses a method for evaluating the ecological toxicity of a biodegradable material by using plant seeds, which comprises the following steps: solution extraction: 1) preparing the degradation material into a powder or block sample for later use; 2) adding an organic solvent into the sample, and then sealing and soaking; 3) oscillating the soaking solution of the sample and the organic solvent, filtering the solution to be used as an experimental group, and setting double controls which are respectively an organic solvent control combined distilled water blank control group; 4) respectively carrying out rotary evaporation and water bath heating on the experimental group solution and the double-control group solution to obtain distillation extraction solutions of the experimental group and the double-control group; 5) dissolving and distilling the extraction solution to respectively obtain experimental extraction solutions of an experimental group and a double control group; (II) germination experiment and index measurement. The method overcomes the defect that the prior art can not effectively evaluate the ecological toxicity of the full-biodegradable material, and can more accurately and more intuitively reflect the ecological toxicity of the full-biodegradable material.
Description
Technical Field
The invention relates to a method for evaluating the ecotoxicity of a full-biodegradable material by using plant seeds, belonging to the field of evaluating the ecotoxicity of the full-biodegradable material.
Background
Plastics are novel materials developed in the beginning of the 20 th century, and become a new industry and are rapidly developed in the 50 th century, and the plastics are widely applied to a plurality of fields such as aviation, automobiles, household appliances, daily necessities, packaging, building materials, medical equipment and the like in the 80 th century. In recent years, the plastic industry in China keeps a rapidly developing situation, the yield and the sales are the first global, and the yield of plastic products accounts for about 20% of the total world yield. Since 2014, the annual output of plastic products in China exceeds 7000 ten thousand tons, and in 2017, 7515.5 ten thousand tons are achieved. However, a large number of disposable plastic packages are used and discarded, which causes serious problems to the environment. For example, disposable foamed plastic lunch boxes and plastic bags are used for containing food, which seriously affects the health of human beings. When the temperature reaches 65 ℃, harmful substances in the disposable foamed plastic tableware permeate into food and can damage the liver, the kidney, the central nervous system and the like of people. The waste agricultural mulching films and plastic bags in the agricultural field remain in the field for a long time, which can affect the absorption of water and nutrients by crops, inhibit the growth and development of the crops and cause the yield reduction of the crops. Survey displayThe average residual film amount of the multi-year film-covered farmland is 71.9-259.1 kg/hm 2 The residual film amount in the soil of the northwest farmland is obviously higher than that of the northwest China and the southwest China. If livestock such as cattle and sheep eat the residual film fragments mixed with crop straws and feed by mistake, gastrointestinal dysfunction can be caused, and even death can be caused. Moreover, the micro-plastics generated in each link enter the atmosphere and water circulation, and seriously harm the human health.
Under the background, research and development and application of fully biodegradable materials and related plastic products prepared by the fully biodegradable materials are imperative. The biodegradable material is a material which is degraded by the action of microorganisms existing in nature under the conditions of nature such as soil and/or sandy soil, and/or under specific conditions such as composting conditions or anaerobic digestion conditions or in an aqueous culture solution, and finally completely degraded into carbon dioxide (CO) 2 ) And/or methane (CH) 4 ) Water (H) 2 O) and mineralized inorganic salts of the elements contained therein, and new biomass materials. As a new material, the safety evaluation of the degradation product of the material is rarely reported, and the invention aims to provide reference for the development and application of the fully biodegradable material by carrying out the biotoxicity evaluation on the fully biodegradable material.
Disclosure of Invention
The invention aims to provide a method for evaluating the ecotoxicity of a full-biodegradable material by using plant seeds.
The technical scheme of the invention is as follows: a method for evaluating the ecological toxicity of a full-biodegradable material by using plant seeds comprises the following steps:
(I) solution extraction
1) Preparing the degradation material into a powder or block sample for later use;
2) adding an organic solvent into the sample, and then sealing and soaking;
3) oscillating the soaking solution of the sample and the organic solvent, filtering the solution to be used as an experimental group, and setting double controls which are an organic solvent control group and a distilled water blank control group respectively;
4) respectively carrying out rotary evaporation and water bath heating on the solutions of the experimental group and the double control group to obtain distilled extraction solutions of the experimental group and the double control group;
5) dissolving and distilling the extraction solution to respectively obtain experimental extraction solutions of an experimental group and a double control group;
(II) Germination test
1) Selecting at least one of monocotyledon and dicotyledon seeds of various families for testing, wherein the seeds are required to be consistent in size, plumpness and same in particle size grade, and the germination rate is over 95 percent;
2) respectively adding equal amounts of the experimental extracting solutions of the experimental group and the double control group into culture dishes with the same specification, and adding equal amounts of distilled water into the culture dishes of the blank group;
3) discharging the same amount of plant seeds in each culture dish, covering the culture dishes, placing the culture dishes in a tissue culture room for culture, adding the same amount of corresponding extracting solution into each culture dish every day, keeping constant weight to supplement evaporated water, and performing germination culture under the condition of keeping the mass concentration of the extracting solution constant;
(III) determination of seed germination percentage and germination vigor
Taking the length of the embryonic roots growing to the length of the seeds as the standard of germination, recording the germination number once every a period of time, and calculating the germination rate and the germination potential of the tobacco seedlings;
(IV) determination of radicle index
Randomly selecting each group of seedlings with the same quantity at set time, cutting off radicles, measuring the height and the length of the seedlings, and calculating the biomass and the ratio of roots to seedlings;
(V) analysis of differences in metabolism of substances
Taking each group of seedlings to perform substance metabolism difference analysis;
(VI) evaluation of results
The germination rates, germination vigor, root-seedling ratio and biomass of the plant seeds of the experimental group and the double control group were compared, and the difference in substance between the experimental group and the control group was evaluated.
Preferably, the extraction solvent is methyl tert-butyl ether.
Preferably, the extraction solution is distilled using ethanol dissolution.
Preferably, the culture conditions in the tissue culture room are as follows: setting the temperature at 26 ℃, setting the relative humidity at more than 85%, taking the length of the radicle growing to the seed as the standard of germination, culturing without light before germination, culturing for 14 days under the light condition with the light-dark ratio of 16:8h after germination.
Preferably, the method for analyzing the difference in metabolism of a substance comprises the following steps:
1) extraction: adding the mixed internal standard solution into a seedling sample, adding the mixed extraction solution, performing vortex, performing normal-temperature ultrasonic extraction, performing vortex again, performing centrifugation, and taking supernatant liquid into a centrifugal tube to obtain an extracting solution for later use;
2) drying: blowing the extracting solution in the centrifugal tube by using nitrogen, adding dichloromethane, and blowing the extracting solution in the centrifugal tube by using nitrogen again;
3) reaction: adding methoxylamine hydrochloride into a centrifuge tube, sealing, performing vortex mixing, reacting at a constant temperature of metal bath at 40 ℃ for 120min, cooling to room temperature, centrifuging, adding BSTFA, sealing, performing vortex mixing, reacting at a constant temperature of metal bath at 70 ℃ for 90min, cooling to room temperature, adding chromatographic pure acetonitrile, sealing, performing vortex mixing, reacting at a constant temperature of metal bath at 81 ℃ for 90min, cooling to room temperature, performing vortex mixing, centrifuging, and taking supernatant to obtain a reaction solution for later use;
4) and (3) detection: and (3) sampling the reaction solution, and performing qualitative and quantitative chromatographic analysis on various substances contained in the solution by using a gas chromatography-mass spectrometry detector.
Preferably, the chromatographic conditions are: a chromatographic column: HP-5MS (60 m.times.250. mu. m.times.0.25 μm) capillary column, injection port temperature: 280 ℃; sample introduction amount: 2 uL; the split ratio is as follows: 20: 1; column flow rate: 1.0 mL/min; temperature rising procedure: keeping at 60 deg.C for 2min, then increasing to 230 deg.C at 5 deg.C/min for 5min, increasing to 290 deg.C at 8 deg.C/min for 23.5min, and operating for 72 min; mass spectrum conditions: ion source temperature: 230 ℃, quadrupole temperature: 150 ℃; ionization energy: 70eV, transmission line temperature: 280 ℃, full scan mass number range 45-600aum, solvent delay: 11.90 min; an acquisition mode: collecting full scanning (Scan); MS spectrum library: the NIST08 library and the Willy08 library.
Preferably, the mixed internal standard solution contains a first internal standard substance and a second internal standard substance, wherein the first internal standard substance is formed by mixing adipic acid, phenyl glucoside and norvaline according to a volume ratio of 10:4:4, and the second internal standard substance is formed by mixing methanol and water according to a volume ratio of 1: 1.
Preferably, the mixed extraction solution is prepared by mixing methanol, chloroform and water according to the volume ratio of 2.5:1: 1.
The invention has the beneficial effects that: the invention treats the full-biodegradable material by the organic solvent, fully dissolves and distills the full-biodegradable material into the extraction solution, uses the extraction solution as an experimental group, uses an organic solvent control group and a distilled water blank control group as a double control group, and evaluates the ecological toxicity of the full-biodegradable material to crops according to the germination experiments treated by the experimental group and the double control group.
The method for evaluating the ecotoxicity of the fully biodegradable material by using the plant seeds overcomes the defect that the ecotoxicity of the fully biodegradable material cannot be effectively evaluated in the prior art, can more accurately and more intuitively reflect the ecotoxicity of the fully biodegradable material, and can further lay a certain foundation for correlation analysis between the fully biodegradable material and ecological pollution and improvement and production of the fully biodegradable material, particularly the fully biodegradable mulching film based on the measurement and analysis results.
Detailed Description
The present invention will be described in detail with reference to the following embodiments in order to make the aforementioned objects, features and advantages of the invention more comprehensible. In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present invention. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein, but rather should be construed as broadly as the present invention is capable of modification in various respects, all without departing from the spirit and scope of the present invention.
Selection of organic solvent: by screening the solvent, except that dichloromethane can dissolve all the degradable materials (films) but cannot dissolve PE, other solvents can not dissolve the degradable materials (films) and PE but dichloromethane is not well filtered, and 1) plastic residues exist after filtering; 2) damaging the filter membrane during filtration; 3) there are problems that some types of degraded materials (membranes) cannot be filtered at all, and the remaining of dissolved degraded materials (membranes) causes serious residue of chromatography instruments.
TABLE 1 comparison of the extraction efficiencies of different solvents for different degradation materials (membranes)
As can be seen from table 1 above, dichloromethane has the greatest extraction efficiency, and the species and content of substances in the extracted and degraded material (membrane) are the highest, but dichloromethane has a slightly poor repeatability, and after filtration, there are disadvantages of plastic residues and easily damaged filter membrane. Therefore, the embodiment of the invention selects the methyl tert-butyl ether as the extraction solvent, the extraction efficiency of the methyl tert-butyl ether is high, the repeatability is good, the filtering is easy, and the main volatile components in the degradation material (membrane) can be better extracted.
The embodiment of the invention provides a method for evaluating the ecotoxicity of a full-biodegradable material by using plant seeds, which comprises the following steps:
(I) solution extraction
1) The degraded material (film) is prepared into powder or small block-shaped samples for standby.
2) Putting the sample into an erlenmeyer flask, adding an organic solvent methyl tert-butyl ether, soaking for 12h, and sealing the erlenmeyer flask to prevent the methyl tert-butyl ether from volatilizing.
3) The sample and organic solvent were shaken for 5 minutes and the solution was filtered into a distillation flask using a double layer gauze as the experimental group, with double controls, each of which was an organic solvent control combined with a distilled water blank control.
4) Respectively carrying out rotary evaporation on the solutions of the experimental group and the double-control group, placing the solutions on a water bath kettle for heating, wherein the temperature of the water bath kettle is 40 ℃, the vacuumizing is 500mbar, the rotating speed is 74rpm, one end of a rotary evaporator device is connected with a flat-bottomed flask (containing waste liquid flowing out in the experimental process), and the other end is connected with the solution (the experimental group and the double-control group) filtered into a distillation flask, so as to obtain the distillation extraction solutions of the experimental group and the double-control group.
5) And dissolving the distilled extraction solution with 1-2 mL of ethanol to obtain experimental extraction solutions of an experimental group and a double-control group respectively.
(II) Germination test
1) At least one of monocotyledon and dicotyledon seeds of various families is selected for testing, the seeds are required to be consistent in size, plumpness and same in particle size grade, and the germination rate is over 95 percent. This example takes the example of a dicotyledonous tobacco seed.
2) The germination test was carried out according to the tobacco seed test protocol (YC/T20-1994). 5.00mL of the experimental extract of the experimental group and the experimental extract of the double control group were added to the same size petri dish (two layers of filter paper on the pad), and the blank group was distilled water.
3) Then, 100 uniform and full tobacco seeds are randomly extracted and arranged on each culture dish according to a certain distance, the culture dishes are covered, the culture dishes are placed in a tissue culture room for culture, the temperature is set to be 26 ℃, the relative humidity is more than 85%, the standard of germination is taken as the embryonic root growth to the seed length, no illumination culture is carried out before germination, the light-dark ratio after germination is 16:8h, the culture is carried out for 14d under the illumination condition, the same amount of corresponding extracting solution is added into each culture dish every day by using a liquid transfer gun, the constant weight is kept to supplement the evaporated moisture, and the germination culture is carried out under the condition that the mass concentration of the extracting solution is kept constant.
(III) determination of seed germination percentage and germination vigor
And taking the length of the growing radicle to the seed as the standard of germination, recording the germination number at intervals of 24 hours, and calculating the germination rate and the germination potential of the tobacco seedling. The faster the germination speed, the more uniform and consistent the germination speed, which indicates that the seed vitality is higher.
Germination rate (GER,%) (germination number of seeds/test seed number) × 100%
Germination vigor (GEP,%) x 100% (number of seeds at peak germination time/number of seeds tested) × (note: 1 germination count per 24h, with the most 1d being peak germination time.)
In the aspect of seed vigor, the germination potential and the germination rate are two important indexes, and the germination rate and the germination potential can also indicate the production performance of the seeds. Therefore, the toxicity of the fully biodegradable material to plants can be effectively evaluated through the germination rate and the germination potential of the seeds.
(IV) radicle index determination
Groups of the same number (e.g., 10) of seedlings were randomly selected at a set time, e.g., day 14, 3 replicates of each treatment were cut, seedling height and root length were determined, and biomass (weight of root and seedling) and root-seedling ratio (height ratio) were calculated.
The root-seedling ratio is equal to root length/seedling height.
(V) analysis of differences in metabolism of substance
Taking each group of seedlings to perform substance metabolism difference analysis, and specifically comprising the following steps:
1) extraction: accurately weighing 50mg of fresh tobacco leaves (50mL of plastic centrifuge tube) of a sample to be tested, adding 40 muL of mixed internal standard (the first internal standard is prepared from 10mg/mL of adipic acid, 8.04mg/mL of phenyl glucoside and 4.9mg/mL of norvaline according to the volume ratio of 10:4:4, the second internal standard is prepared from methanol and water according to the volume ratio of 1: 1), adding 3mL of mixed extracting solution (prepared from methanol, trichloromethane and water according to the volume ratio of 2.5:1: 1), homogenizing the fresh tobacco leaves for 1 minute for extraction, carrying out vortex for 1 minute, carrying out ultrasonic extraction at normal temperature for 40 minutes (vortex for 1 time halfway and vortex for 1 minute), finally carrying out vortex for 1 minute again, placing at 3000 plus 5000rpm for centrifugation for 5 minutes, carefully taking 600 muL of supernatant liquid to be placed in a 2mL of centrifuge tube, and obtaining the extracting solution for later use.
2) Drying: blowing the extracting solution in the centrifugal tube by using nitrogen, adding dichloromethane, and blowing the extracting solution in the centrifugal tube by using nitrogen again. Specifically, nitrogen is firstly blown for one half hour to two hours, the liquid level descending condition is observed every 20-30min in the middle, the height of a blowing needle is properly adjusted according to the actual condition, the gas flow (the flow is not too large, the liquid is not required to splash out of a centrifuge tube or be attached to the needle), 100 mu L of dichloromethane is added after no obvious liquid is observed in the centrifuge tube, the height of the needle is properly adjusted from the first centrifuge tube immediately after the dichloromethane is sequentially added, the blowing time is kept for one half hour, the dichloromethane can be recovered, and the dichloromethane is sealed by a sealing film for low-temperature storage (if water gas exists on the wall of the centrifuge tube, the dichloromethane is wiped by paper). The nitrogen is used in an amount of substantially 3 to 4 times per pass.
3) Reaction: drying fresh tobacco leaf with 600 μ L of extractive solution with nitrogen, drying (drying, storing at 4 deg.C, sealing with sealing film, recovering to room temperature before use, wiping off water vapor, opening for the next step), adding 40 μ L of 25-30 (preferably 30) mg/mL methoxylamine hydrochloride, adding 1, covering with 1 (prepared with dry pyridine, prepared with large bottle of pyridine), sealing with sealing film, mixing by vortex for 5min, reacting at 40 deg.C for 120min (vortex for 1 time at 60min for about 30s), cooling to room temperature, centrifuging slightly, and adding 200 μ LBS TFA (TMCSL 1%). Adding 1, covering 1, sealing with a sealing film, performing vortex mixing for 30s, performing reaction at 70 ℃ for 90min (slightly swirling for 10s when 45 min) in a constant-temperature metal bath, cooling to room temperature, adding 90 mu L of chromatographic pure acetonitrile, performing vortex mixing for 30s after sealing with the sealing film, performing reaction at 81 ℃ for 90min (slightly swirling for 10s when 45 min) in the constant-temperature metal bath, cooling to room temperature, performing vortex mixing, centrifuging for 3min, and taking 100 mu L of supernatant to obtain a reaction solution for later use.
4) And (3) detection: and (3) sampling the reaction solution in a 250-mu-L liner tube, and performing qualitative and quantitative chromatographic analysis on various substances contained in the solution by using a gas chromatography-mass spectrometry detector. Preferably, the chromatographic conditions are: a chromatographic column: HP-5MS (60 m.times.250. mu.m.times.0.25 μm) capillary chromatography column, injection port temperature: 280 ℃; sample introduction amount: 2 uL; the split ratio is as follows: 20: 1; column flow rate: 1.0 mL/min; temperature rising procedure: keeping at 60 deg.C for 2min, then increasing to 230 deg.C at 5 deg.C/min for 5min, increasing to 290 deg.C at 8 deg.C/min for 23.5min, and operating for 72 min; mass spectrum conditions: ion source temperature: 230 ℃, quadrupole temperature: 150 ℃; ionization energy: 70eV, transmission line temperature: 280 ℃, full scan mass number range 45-600aum, solvent delay: 11.90 min; an acquisition mode: collecting full scanning (Scan); MS spectrum library: the NIST08 library and the Willy08 library.
(VI) evaluation of results
According to the results of multiple experiments, when the germination rate, the germination vigor, the biomass and the ratio of roots to seedlings of the tobacco seeds in the experimental group are less than 90% of the corresponding values of the control group, the fully biodegradable material has toxic influence on the plant seeds, and when the ratio is more than 90%, the fully biodegradable material does not have toxic influence.
The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the present invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.
Claims (2)
1. A method for evaluating the ecotoxicity of a full-biodegradable material by using plant seeds is characterized by comprising the following steps:
(I) solution extraction
1) Preparing the degradation material into a powder or block sample for later use;
2) adding an organic solvent into the sample, and then sealing and soaking, wherein the organic solvent is methyl tert-butyl ether;
3) oscillating the soaking solution of the sample and the organic solvent, filtering the solution to be used as an experimental group, and setting double controls which are an organic solvent control group and a distilled water blank control group respectively;
4) respectively carrying out rotary evaporation and water bath heating on the solutions of the experimental group and the double control group to obtain distilled extraction solutions of the experimental group and the double control group;
5) dissolving and distilling the extraction solution by adopting ethanol to respectively obtain experimental extraction solutions of an experimental group and a double control group;
(II) Germination test
1) Selecting at least one of monocotyledon and dicotyledon seeds of various families for testing, wherein the seeds are required to be consistent in size, plumpness and same in particle size grade, and the germination rate is over 95 percent;
2) respectively adding equal amounts of the experimental extracting solutions of the experimental group and the double control group into culture dishes with the same specification, and adding equal amounts of distilled water into the culture dishes of the blank group;
3) discharging the same amount of plant seeds in each culture dish, covering the culture dishes, placing the culture dishes in a tissue culture room for culture, adding the same amount of corresponding extracting solution into each culture dish every day, keeping constant weight to supplement evaporated water, and performing germination culture under the condition of keeping the mass concentration of the extracting solution constant;
(III) determination of seed germination percentage and germination vigor
Taking the length of the embryonic roots growing to the length of the seeds as the standard of germination, recording the germination number once every a period of time, and calculating the germination rate and the germination vigor;
(IV) determination of radicle index
Randomly selecting each group of seedlings with the same quantity at set time, cutting off radicles, measuring the height and the length of the seedlings, and calculating the biomass and the ratio of roots to seedlings;
(V) analysis of differences in metabolism of substance
Taking each group of seedlings to perform substance metabolism difference analysis, wherein the substance metabolism difference analysis method comprises the following steps:
1) extraction: adding the mixed internal standard solution into a seedling sample, adding the mixed extraction solution, performing vortex, performing normal-temperature ultrasonic extraction, performing vortex again, performing centrifugation, and taking supernatant liquid into a centrifugal tube to obtain an extracting solution for later use;
2) drying: blowing the extracting solution in the centrifugal tube by using nitrogen, adding dichloromethane, and blowing the extracting solution in the centrifugal tube by using nitrogen again;
3) reaction: adding methoxylamine hydrochloride into a centrifuge tube, sealing, performing vortex mixing, reacting at a constant temperature of metal bath at 40 ℃ for 120min, cooling to room temperature, centrifuging, adding BSTFA, sealing, performing vortex mixing, reacting at a constant temperature of metal bath at 70 ℃ for 90min, cooling to room temperature, adding chromatographic pure acetonitrile, sealing, performing vortex mixing, reacting at a constant temperature of metal bath at 81 ℃ for 90min, cooling to room temperature, performing vortex mixing, centrifuging, and taking supernatant to obtain a reaction solution for later use;
4) and (3) detection: sampling the reaction solution, and performing qualitative and quantitative chromatographic analysis on various substances contained in the solution by using a gas chromatography-mass spectrometry detector;
the mixed internal standard solution contains a first internal standard substance and a second internal standard substance, wherein the first internal standard substance is formed by mixing adipic acid, phenyl glucoside and norvaline according to a volume ratio of 10:4:4, and the second internal standard substance is formed by mixing methanol and water according to a volume ratio of 1: 1; the mixed extraction solution is prepared by mixing methanol, trichloromethane and water according to the volume ratio of 2.5:1: 1;
the chromatographic conditions are as follows: a chromatographic column: HP-5MS capillary chromatographic column, 60m × 250 μm × 0.25 μm, injection port temperature: 280 ℃; sample introduction amount: 2 mu L of the solution; the split ratio is as follows: 20: 1; column flow rate: 1.0 mL/min; temperature rising procedure: keeping at 60 deg.C for 2min, then increasing to 230 deg.C at 5 deg.C/min for 5min, increasing to 290 deg.C at 8 deg.C/min for 23.5min, and operating for 72 min; mass spectrum conditions: ion source temperature: 230 ℃, quadrupole temperature: 150 ℃; ionization energy: 70eV, transmission line temperature: 280 ℃, full scan mass number range 45-600aum, solvent delay: 11.90 min; an acquisition mode: collecting full scanning; MS spectrum library: the NIST08 and Willy08 libraries;
(VI) evaluation of results
And (3) comparing the germination rates, the germination potentials, the root-seedling ratios and the biomass of the plant seeds of the experimental group and the double control groups, and simultaneously evaluating the substance difference between the experimental group and the control groups.
2. The method for evaluating the ecotoxicity of the whole biodegradable material using the plant seeds as claimed in claim 1, wherein the culture conditions in the tissue culture room are: setting the temperature at 26 ℃, setting the relative humidity at more than 85%, taking the length of the radicle growing to the seed as the standard of germination, culturing without light before germination, culturing for 14 days under the light condition with the light-dark ratio of 16:8h after germination.
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| CN113419052A (en) * | 2021-05-21 | 2021-09-21 | 江苏大学 | Method for evaluating micro-plastic pollution level based on plant physiological indexes |
| CN115015450B (en) * | 2022-05-26 | 2024-05-14 | 贵州省烟草科学研究院 | Method for analyzing metabolites in soil by microwave derivatization-quasi-target gas chromatography-mass spectrometry |
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| CN107860877B (en) * | 2017-11-02 | 2020-06-02 | 湖南大学 | Method for evaluating the phytotoxicity of antibiotics |
| CN109164189A (en) * | 2018-10-17 | 2019-01-08 | 河北科技大学 | γ-phytotoxic the evaluation method of hexabromocyclododecane enantiomer |
| CN110261590B (en) * | 2019-03-15 | 2022-10-04 | 暨南大学 | Method for evaluating phytotoxicity of phthalate |
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