CN111208029A - Method for evaluating ecotoxicity of organic additive in full-biodegradable mulching film by applying water culture experiment - Google Patents
Method for evaluating ecotoxicity of organic additive in full-biodegradable mulching film by applying water culture experiment Download PDFInfo
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Abstract
The invention discloses a method for evaluating the ecotoxicity of an organic additive in a full-biodegradable mulching film by applying a water culture experiment, which comprises the following steps: solution extraction: s1, cutting the full-biodegradable mulching film into pieces to obtain samples for later use; s2, adding the internal standard solution and the extraction solution into the sample, performing sealed extraction by adopting a microwave extraction method, performing vortex extraction, and filtering to obtain an extraction liquid; s3, adding a mixed solution of ultrapure water, an extraction solvent and a dispersion solvent into the extraction liquid, shaking up, adding sodium chloride, performing vortex-assisted dispersion liquid micro-extraction, centrifuging, and taking a lower-layer solution to obtain an organic additive solution; (II) water culture experiment; and (III) carrying out sample injection analysis on the organic additive. The invention adopts microwave-assisted extraction to effectively improve the recovery rate of the organic additive in the fully biodegradable mulching film and reduce the extraction time, and meanwhile, the dispersion liquid-liquid microextraction can effectively remove the interference of a large amount of non-target compounds, so that the organic additive of the extract is purer and the ecological toxicity of the organic additive can be truly reflected.
Description
Technical Field
The invention relates to a method for evaluating the ecotoxicity of an organic additive in a full-biodegradable mulching film by applying a water culture experiment, belonging to the field of analysis of the ecotoxicity of the organic additive in the full-biodegradable mulching film.
Background
The full-biodegradable mulching film is a novel mulching film which can be completely degraded by microorganisms in 1-2 plant growth cycles, the degradation principle is that the surface of the mulching film is adhered by the microorganisms, the microorganisms use organic carbon in polyester as a nutrient carbon source, under the action of enzyme, high molecules are decomposed into fragments with lower molecular weight, the fragments are absorbed or consumed by the microorganisms, carbon dioxide, water, mineral salt and new biomass are formed through metabolism, and finally the carbon dioxide, water, mineral salt and new biomass are completely absorbed by the nature. The technology overcomes the pollution of polyethylene or polypropylene residual films from the source and has wide application prospect in agricultural production. The main components of the full-biodegradable mulching film are full-biodegradable resin (polylactic acid (PLA), butylene terephthalate-butylene adipate (PBAT), polybutylene succinate (PBS), Polycaprolactone (PCL) and the like), and the full-biodegradable mulching film has no toxic unit in a chemical structure, so that the environment is nontoxic, and the material composition and content in the full-biodegradable mulching film are less researched.
The full-biodegradable mulch additive is a compound which is intentionally or unintentionally added into a polymer in the production process of the mulch, so that the material can achieve the required physical and chemical properties, and the performance and the applicability of the material are improved to meet the production and use requirements. The additives can be classified into organic (antioxidant, light stabilizer, lubricant, etc.) or inorganic (silicate, titanate, aluminate, etc.) additives according to the composition, wherein the organic additives mainly comprise various compounds such as acids (containing benzene ring), amines, fatty amides, esters, phenols, etc., and some organic additives have obvious toxic units and certain acute and chronic toxicity. The substances are bound to be transferred into soil and underground water along with the use of the mulching film, so that the environment is polluted, and the human health is harmed. Therefore, the development of toxicity evaluation of the main organic additives in the fully biodegradable mulching film has very important significance for safe use of the fully biodegradable mulching film.
However, the ecotoxicity of organic additives in fully biodegradable mulch cannot be effectively evaluated at present, mainly because: firstly, the full-biodegradable mulching film is used as a new material, and no effective method for evaluating the safety of degradation products of the full-biodegradable mulching film exists at present; secondly, the prior art also does not disclose a method for extracting the main organic additive in the full-biodegradable mulching film. The existing literature is mainly used for detecting and analyzing main organic additives in non-degradable plastic packages. In the aspect of pretreatment extraction, methods such as ultrasonic extraction, microwave-assisted extraction, static/dynamic headspace analysis, solid-phase microextraction and the like are mainly utilized, wherein the ultrasonic extraction efficiency is low, and the time is long; static/dynamic headspace analysis, solid phase microextraction methods are generally only suitable for analyzing volatile components; the microwave-assisted extraction efficiency is high, the time is short, but a large amount of non-target compounds are extracted, the toxic interference on organic additives is large, and pollution is easy to generate and target chromatographic peaks are easy to interfere when a chromatographic system is evaluated.
Disclosure of Invention
The technical problems to be solved by the invention are as follows: the method adopts microwave-assisted extraction to effectively improve the recovery rate of the organic additive in the fully biodegradable mulching film and reduce the extraction time, and meanwhile, the dispersion liquid-liquid micro-extraction can effectively remove the interference of a large amount of non-target compounds, so that the organic additive of the extract is purer, the ecological toxicity of the organic additive can be truly reflected, and the defects in the prior art are overcome.
The technical scheme of the invention is as follows: a method for evaluating the ecological toxicity of an organic additive in a full-biodegradable mulching film by applying a water culture experiment comprises the following steps:
(I) solution extraction
S1, sample pretreatment: shearing the full-biodegradable mulching film to obtain a sample for later use;
s2, extracting organic additives: adding the internal standard solution and the extraction solution into the sample, performing sealed extraction by adopting a microwave extraction method, performing vortex extraction, and filtering to obtain an extraction liquid for later use;
s3, micro-extraction purification of organic additives: adding a mixed solution of ultrapure water, an extraction solvent and a dispersion solvent into the extract, shaking up, adding sodium chloride, performing vortex-assisted dispersion liquid-liquid microextraction, centrifuging, and taking a lower-layer solution to obtain an organic additive solution;
(II) Water culture experiment
1) Adding nutrient solution into organic additive solution as experimental group, adding equal amount of nutrient solution into distilled water as control group, wherein the volumes of the organic additive solution and the distilled water are the same;
2) respectively transferring the solutions of the experimental group and the control group which are soaked overnight into two water culture boxes, and adjusting the pH values of the solutions of the experimental group and the control group to 6.6-6.9;
3) placing the same number of plant seeds exposed to the white in each water culture box, and leaving one plant seed with the best growth vigor after the seeds grow out of true leaves;
4) adding equal amount of nutrient solution into each water culture tank at intervals to a water level line, keeping constant weight to supplement evaporated water, and managing according to a conventional mode;
5) randomly selecting the same number of seedlings in each group at a set time, measuring root system and stem leaf biomass of the seedlings in the experimental group and the seedlings in the control group, and comparing and judging the toxicity of the organic additive solution;
(III) analysis of organic additives by sample injection
And (3) sampling organic additive solution, and performing qualitative and quantitative chromatographic analysis on various organic additives by using a gas chromatography-mass spectrometry detector.
Preferably, the internal standard solution in step S2 is a phenylethyl acetate solution, and the extraction solution is a methanol solution.
Preferably, the microwave extraction method in step S2 adopts a closed microwave-assisted extraction system, and the microwave extraction conditions are as follows: the microwave power is 800w, the temperature is 80 ℃, and the extraction time is 20 min.
Preferably, the mixed solution of the extraction solvent and the dispersion solvent in step S3 is a mixed solution of carbon tetrachloride and acetone.
Preferably, the chromatographic conditions in the organic additive sample injection analysis are as follows: capillary chromatographic column, temperature programming, sample inlet temperature of 280 ℃, split-flow sample injection, split-flow ratio: 5:1, carrier gas: high purity helium, constant flow rate: 1.0mL/min, sample size: 1.0 μ L; mass spectrometry conditions were electron impact ion source (EI source), ion source temperature: 230 ℃, quadrupole temperature: 150 ℃, mass transfer line temperature: 280 ℃, ion source voltage: 70eV, solvent delay of 7min, qualitative full scan in a scanning range of 50-500 amu, and quantitative selective ion scan.
Preferably, the capillary chromatographic column is a 5% phenyl-95% dimethylpolysiloxane stationary phase, the specification is 60m × 0.25mm, the thickness of the film is 0.25 μm, the temperature programming condition is that the initial temperature is 60 ℃, and the temperature is kept for 2 min; raising the temperature to 230 ℃ at a speed of 15 ℃/min, and keeping the temperature for 5 min; raising the temperature to 280 ℃ at a speed of 5 ℃/min and keeping the temperature for 20 min; raising the temperature to 290 ℃ at a speed of 3 ℃/min, and keeping the temperature for 5 min.
Preferably, the organic additives are qualitatively analyzed by retention time, NIST08, Wiley08, retention index or standard substance, and quantitatively analyzed by internal standard method.
Preferably, the quantification is performed by absolute quantification and relative quantification respectively using phenylethyl acetate as an internal standard.
The invention has the beneficial effects that: the invention establishes a method for extracting main organic additives in a full-biodegradable mulching film based on microwave-assisted extraction combined with a dispersion liquid-liquid microextraction technology and carrying out a water culture experiment on the organic additives to evaluate the ecological toxicity of the organic additives, and compared with the prior art, the method has the advantages that: 1. the microwave-assisted extraction can effectively improve the extraction efficiency and reduce the extraction time; 2. the interference of non-target compounds can be effectively reduced by the liquid-liquid microextraction of the dispersion liquid, so that the organic additive of the extract is purer and the ecological toxicity of the organic additive can be truly reflected; 3. by combining microwave-assisted extraction and dispersion liquid-liquid microextraction, 71 main organic additives in the fully biodegradable mulching film can be analyzed simultaneously by adopting a gas chromatography-mass spectrometry detection method, so that the sensitivity of the method to additive detection can be improved; 4. the method has the characteristics of simplicity, rapidness and good stability, and can effectively improve the data accuracy of sample detection.
The method for evaluating the ecotoxicity of the organic additive in the fully biodegradable mulching film by applying the water culture experiment overcomes the defect that the prior art cannot effectively evaluate the ecotoxicity of the organic additive in the fully biodegradable mulching film, can more accurately and more intuitively reflect the ecotoxicity of the organic additive in the fully biodegradable mulching film, and can further lay a certain foundation for the correlation analysis between the organic additive of the fully biodegradable mulching film and ecological pollution and the improved production of the fully biodegradable mulching film based on the measurement and analysis results.
Drawings
FIG. 1 is a gas chromatography-mass spectrometry chromatogram of 14 organic additive standards;
FIG. 2 is a gas chromatography-mass spectrometry chromatogram of 71 main organic additives in the fully biodegradable mulch.
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.
Example 1
The method for evaluating the ecotoxicity of the organic additive in the full-biodegradable mulch film by applying the water culture experiment comprises the following steps of:
(I) solution extraction
S1, sample pretreatment: and shearing the full-biodegradable mulching film by using scissors, and uniformly mixing to obtain a sample for later use.
S2, extracting organic additives: weighing 100mg of sample in a microwave extraction tank, adding 5 mu L of phenethyl acetate internal standard solution with the concentration of 1.852mg/mL, adding 1.7mL of methanol extraction solution, covering and sealing, placing in a sealed microwave-assisted extraction system for extraction under the microwave extraction conditions that the microwave power and the temperature are respectively set as 800W and 80 ℃, the extraction time is 20min, carrying out vortex for 60s after microwave, and passing the extract through a 0.22 mu m microporous membrane in a 2mL centrifuge tube to obtain the extract for later use.
S3, micro-extraction purification of organic additives: adding 5mL of ultrapure water into a 10mL centrifuge tube, transferring 0.8mL of extract liquor, adding a mixed extraction solvent and a dispersion solvent of 80 μ L of carbon tetrachloride and 200 μ L of acetone, shaking up, adding 0.4g of sodium chloride, performing vortex-assisted dispersion liquid microextraction for 60s, centrifuging at 4000rpm for 5min, and taking a lower layer solution to obtain an organic additive solution.
(II) Germination test
1) Adding nutrient solution into organic additive solution as experimental group, adding equal amount of nutrient solution into distilled water as control group, wherein the volumes of organic additive solution and distilled water are the same. The nutrient solution may be Kawasaki nutrient solution.
2) And transferring the solutions of the experimental group and the control group after being soaked overnight into two water culture boxes respectively, and adjusting the pH values of the solutions of the experimental group and the control group to 6.6-6.9. The pH of the nutrient solution can be adjusted (by adding acid or alkali) by using a pH meter.
3) The same number of exposed monocotyledon or dicotyledon seeds are placed in each water culture box, and lettuce is taken as an example in the embodiment. After the seeds of the lettuce grow out of the true leaves, one single plant is left with the growth vigor more consistent with the integral growth vigor of the water culture box. The seeds used are consistent in size, identical in plumpness and particle size grade, and the germination rate is over 95%. Specifically, the sponge can be placed into the solution of the hydroponic box in advance to be soaked for 10 minutes, then water is squeezed, the sponge is kept in a wet state (the purpose is to facilitate moisture absorption after the seeds are placed in the box, and meanwhile, the seeds cannot germinate because the seeds are corroded or anaerobic due to excessive water), and the seeds exposed to the white are lightly clamped by tweezers and placed into the sponge. Put into the sponge and down drag the sponge with tweezers after, unify the sponge height, ensure that the sponge bottom contacts the solution in the water planting case, guarantee that the sponge can absorb water, improve the seed germination percentage. Two plant seeds exposed to the white are placed in each sponge, after the true leaves grow out, one plant with the growth vigor consistent with the overall growth vigor of the water culture box is reserved, and the uniformity of the seedlings is improved. The machine is plugged in a power supply, a switch is turned on, and Beijing time 7 is set: 00 (in this case, during sprout cultivation, no visible light is required). The nutrient solution in the water culture box is gradually reduced along with the growth of the lettuce, so the nutrient solution needs to be supplemented to the water level line every 10 days. And (3) when the seeds germinate, entering an illumination mode, wherein the light-dark ratio is 12: 12.
4) adding equal amount of nutrient solution into the water culture tank at intervals to reach water level, maintaining constant weight to supplement water evaporated or absorbed by plant, and managing according to conventional mode.
5) Randomly selecting lettuce of the same quantity in each group at set time, measuring root system and stem leaf biomass of the lettuce in the experimental group and the control group, and comparing and judging the toxicity of the organic additive solution;
specifically, the same number (for example, 10) of lettuce in each group is randomly selected at a set time, for example, on 30 th, 60 th and 90 th days, each treatment is repeated 3 times, plant roots and stems and leaves are cut off, and the biomass of the roots and stems and leaves of the lettuce in the experimental group and the control group is measured.
According to multiple experimental results, compared with corresponding values of a control group, the biomass of roots and stems and leaves of tobacco seeds in an experimental group is less than 90% of the biomass of roots and stems and leaves of tobacco seeds in the full-biodegradable mulching film, and the biomass of roots and stems and leaves of tobacco seeds in the experimental group has toxic influence on plant hydroponic growth, and more than 90% of the biomass of roots and leaves of tobacco seeds in the experimental group does not.
(III) analysis of organic additives by sample injection
And (3) taking the organic additive solution into a 100 mu L micro-sample bottle by using a 100 mu L manual sample injection valve injector, and carrying out chromatographic analysis by using a gas chromatography-mass spectrometry detector. The chromatographic conditions are 5% phenyl-95% dimethyl polysiloxane stationary phase capillary chromatographic column, the specification is 60m × 0.25mm, the film thickness is 0.25 μm, the temperature programming condition is initial temperature 60 ℃, and the temperature is kept for 2 min; raising the temperature to 230 ℃ at a speed of 15 ℃/min, and keeping the temperature for 5 min; raising the temperature to 280 ℃ at a speed of 5 ℃/min, and keeping the temperature for 20 min; raising the temperature to 290 ℃ at the speed of 3 ℃/min, keeping the temperature for 5min, and carrying out split injection at the injection inlet temperature of 280 ℃, wherein the split ratio is as follows: 5:1, carrier gas: high purity helium (99.999%), constant flow rate: 1.0mL/min, sample size: 1.0 μ L; mass spectrometry conditions were electron impact ion source (EI source), ion source temperature: 230 ℃, quadrupole temperature: 150 ℃, mass transfer line temperature: 280 ℃, ion source voltage: 70e V, solvent delay of 7min, qualitative full Scan (Scan mode), Scan range of 50-500 amu, quantitative selective ion Scan (SIM mode), and selective ion shown in Table 1.
TABLE 1 name of 71 organic additives in full-biodegradable mulch film, CAS, retention index, qualitative and quantitative ion of mass spectrum and identification matching degree
The results of the standard sample and the sample are shown in fig. 1 and fig. 2 by using all 71 main organic additives for qualitative and quantitative determination through gas chromatography-mass spectrometry. Among them, 14 organic additives, 2, 6-di-t-butyl-p-cresol, 1, 6-dioxadecane-7, 12-dione, 2, 4-trimethyl-1, 3-pentanediol diisobutyrate, 2,6,6, 8-tetramethyltricyclo [5.3.1.0] undecane-8-ol, n-heptadecane, 3, 5-di-t-butyl-4-hydroxybenzaldehyde, phenanthrene, dibutyl phthalate, hexadecanenitrile, octyl p-methoxycinnamate, diethylhexyl adipate, eicosamide, 2-hydroxy-4-n-octyloxybenzophenone, and trans-squalene, were absolutely quantified by an internal standard curve method, and 57 organic additives without standards were relatively quantified. Qualitative analysis was performed by retention time, NIST08, Wiley08, retention index or standard substance, and quantitative analysis was performed by internal standard method, respectively. And quantitative determination is carried out by taking phenethyl acetate as an internal standard for absolute quantification and relative quantification respectively. Using a standard curve to absolutely quantify 14 organic additives with standard substances; the other 57 organic additives were relatively quantified using the internal standard phenylethyl acetate at a correction factor F ═ 1.
Preparing mixed standard solution (mother solution) of 14 organic additives with corresponding concentration, wherein 1, 6-dioxacyclodecane-7, 12-dione, hexadecanenitrile, dibutyl phthalate, eicosamide, 2, 6-di-tert-butyl-p-cresol and trans-squalene are all prepared and dissolved by methanol, transferred into a 25.00mL volumetric flask, and subjected to constant volume to a scale mark and shaking to obtain 0.10mg/mL standard solution; 2,2, 4-trimethyl-1, 3-pentanediol diisobutyrate, 2,6,6, 8-tetramethyltricyclo [5.3.1.0] undecane-8-ol, n-heptadecane, 3, 5-di-tert-butyl-4-hydroxybenzaldehyde, phenanthrene, octyl p-methoxycinnamate, and 2-hydroxy-4-n-octyloxybenzophenone were prepared as a 0.05mg/mL standard solution by the above method; diethyl hexyl adipate was prepared as a 0.5mg/mL standard solution as described above, and the mother liquors were mixed and diluted with methanol to 5 concentration-gradient mixed standards.
Performing micro-extraction, chromatographic qualitative analysis and quantitative analysis according to the conditions of the S3 (standard chromatogram is shown in figure 1), performing regression analysis by taking the ratio of the peak area of the target compound to the peak area of the internal standard substance as an ordinate (y) and the ratio of the amount of the target compound to an internal scalar (9.26 mu g) as an abscissa (x) to obtain a regression equation and related parameters, calculating the lowest detection Limit (LOD) of the method by using the standard solution with the lowest concentration according to the signal-to-noise ratio of 3 (S/N-3), adding the standard solutions with two concentration gradients into the fully biodegradable mulch film, extracting, performing derivatization and chromatographic analysis, and calculating the recovery rate. The recovery rate was calculated as the additive content of the sample after addition minus the additive content of the blank divided by the additive content, with the addition concentrations being approximately 1/2 times and 1 time the additive content of the blank, respectively, with the precision of the method being 5 repetitions of the low and high concentration additions within the same day, for the day precision, and 5 repetitions of the same sample within 5 days for the day precision. The results are shown in tables 2 and 3.
TABLE 2 Standard Curve, Linear Range, LOD and Linear correlation coefficient for the major organic additives
TABLE 3 recovery rate of addition of main organic additives, precision in and between days
The results show that: the linear correlation coefficients of the 14 organic additives with the standard substances are all larger than 0.999, the detection sensitivity of each organic additive is improved through the optimization of dispersion liquid-liquid microextraction, and the LOD of all the organic additives is smaller than 0.0586 mu g/g. The recovery rate of each organic additive is between 92.3% and 109.7%, the recovery rate is good, the precision ranges of the daily precision and the daily precision are between 2.17% and 13.69% and 3.51% and 14.01%, and the method meets the strict quantitative requirement.
The contents of the embodiments show that the invention establishes a method for extracting the main organic additives in the full-biodegradable mulching film based on the microwave-assisted extraction combined with the dispersion liquid-liquid microextraction technology and carrying out the water culture experiment on the organic additives to evaluate the ecological toxicity of the organic additives, and compared with the prior art, the method has the advantages that: 1. the microwave-assisted extraction can effectively improve the extraction efficiency and reduce the extraction time; 2. the interference of non-target compounds can be effectively reduced by the liquid-liquid microextraction of the dispersion liquid, so that the organic additive of the extract is purer and the ecological toxicity of the organic additive can be truly reflected; 3. by combining microwave-assisted extraction and dispersion liquid-liquid microextraction, 71 main organic additives in the fully biodegradable mulching film can be analyzed simultaneously by adopting a gas chromatography-mass spectrometry detection method, so that the sensitivity of the method to additive detection can be improved; 4. the method has the characteristics of simplicity, rapidness and good stability, and can effectively improve the data accuracy of sample detection.
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 (8)
1. A method for evaluating the ecotoxicity of an organic additive in a full-biodegradable mulching film by applying a water culture experiment is characterized by comprising the following steps of:
(I) solution extraction
S1, sample pretreatment: shearing the full-biodegradable mulching film to obtain a sample for later use;
s2, extracting organic additives: adding the internal standard solution and the extraction solution into the sample, performing sealed extraction by adopting a microwave extraction method, performing vortex extraction, and filtering to obtain an extraction liquid for later use;
s3, micro-extraction purification of organic additives: adding a mixed solution of ultrapure water, an extraction solvent and a dispersion solvent into the extract, shaking up, adding sodium chloride, performing vortex-assisted dispersion liquid-liquid microextraction, centrifuging, and taking a lower-layer solution to obtain an organic additive solution;
(II) Water culture experiment
1) Adding nutrient solution into organic additive solution as experimental group, adding equal amount of nutrient solution into distilled water as control group, wherein the volumes of the organic additive solution and the distilled water are the same;
2) respectively transferring the solutions of the experimental group and the control group which are soaked overnight into two water culture boxes, and adjusting the pH values of the solutions of the experimental group and the control group to 6.6-6.9;
3) placing the same number of plant seeds exposed to the white in each water culture box, and leaving one plant seed with the best growth vigor after the seeds grow out of true leaves;
4) adding equal amount of nutrient solution into each water culture tank at intervals to a water level line, keeping constant weight to supplement evaporated water, and managing according to a conventional mode;
5) randomly selecting the same number of seedlings in each group at a set time, measuring root system and stem leaf biomass of the seedlings in the experimental group and the seedlings in the control group, and comparing and judging the toxicity of the organic additive solution;
(III) analysis of organic additives by sample injection
And (3) sampling organic additive solution, and performing qualitative and quantitative chromatographic analysis on various organic additives by using a gas chromatography-mass spectrometry detector.
2. The method for evaluating the ecotoxicity of the organic additive in the fully biodegradable mulch film according to claim 1, wherein the internal standard solution is a phenylethyl acetate solution and the extraction solution is a methanol solution in step S2.
3. The method for evaluating the ecotoxicity of the organic additive in the fully biodegradable mulch film by applying the hydroponic experiment as claimed in claim 1, wherein the microwave extraction method in the step S2 adopts a closed microwave-assisted extraction system, and the microwave extraction conditions are as follows: the microwave power is 800w, the temperature is 80 ℃, and the extraction time is 20 min.
4. The method for evaluating the ecotoxicity of the organic additive in the fully biodegradable mulch film according to claim 1, wherein the mixed solution of the extraction solvent and the dispersion solvent in the step S3 is a mixed solution of carbon tetrachloride and acetone.
5. The method for evaluating the ecotoxicity of the organic additive in the fully biodegradable mulch film by applying the hydroponic experiment according to claim 1, wherein the chromatographic conditions in the organic additive sample injection analysis are as follows: capillary chromatographic column, temperature programming, sample inlet temperature of 280 ℃, split-flow sample injection, split-flow ratio: 5:1, carrier gas: high purity helium, constant flow rate: 1.0mL/min, sample size: 1.0 μ L; mass spectrometry conditions were electron impact ion source (EI source), ion source temperature: 230 ℃, quadrupole temperature: 150 ℃, mass transfer line temperature: 280 ℃, ion source voltage: 70eV, solvent delay of 7min, qualitative full scan in a scanning range of 50-500 amu, and quantitative selective ion scan.
6. The method for evaluating the ecotoxicity of the organic additive in the fully biodegradable mulch film by applying the water culture experiment according to claim 5, wherein the capillary chromatographic column is 5% phenyl-95% dimethylpolysiloxane stationary phase, the specification is 60m x 0.25mm, the film thickness is 0.25 μm, the temperature programming condition is 60 ℃ at the initial temperature, and the temperature is kept for 2 min; raising the temperature to 230 ℃ at a speed of 15 ℃/min, and keeping the temperature for 5 min; raising the temperature to 280 ℃ at a speed of 5 ℃/min and keeping the temperature for 20 min; raising the temperature to 290 ℃ at a speed of 3 ℃/min, and keeping the temperature for 5 min.
7. The method for evaluating the ecotoxicity of the organic additives in the fully biodegradable mulch film according to claim 5 using hydroponic experiments wherein the organic additives are qualitatively analyzed by retention time, NIST08, Wiley08, retention index or standard substance and quantitatively analyzed by internal standard method.
8. The method for evaluating the ecotoxicity of the organic additive in the fully biodegradable mulch film by applying the hydroponic experiment as claimed in claim 5, wherein the quantification is performed by taking phenethyl acetate as an internal standard for absolute quantification and relative quantification respectively.
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