CN108593401B - Method for separating submicron-order micro-plastic in water environment or marine products - Google Patents
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Abstract
A method for separating submicron order micro plastic comprises the following steps: adding sodium dodecyl sulfate into a sample solution, ultrasonically dispersing uniformly, standing, then dividing into a suspension layer, a solution layer and a precipitation layer from top to bottom, removing the middle solution layer through siphoning, and collecting the residue; adding 2-3 volume times of absolute ethyl alcohol into the collected residues, standing after ultrasonic treatment to separate the mixture into two layers, removing the upper layer solution through siphoning, and collecting the lower layer substance; placing the collected lower-layer substance in a flat bottom vessel, adding 1-3 volume times of absolute ethyl alcohol, uniformly mixing, standing until the lower-layer substance is divided into two layers, absorbing the upper-layer liquid, and naturally airing to separate the upper-layer liquid into submicron-grade micro plastic in a sample liquid; the method can quickly and accurately separate and extract the submicron plastic particles in the sample, does not need suction filtration, centrifugation and drying, has simple operation and low cost, and provides technical support for subsequent accurate quantitative detection.
Description
(I) technical field
The invention relates to a method for separating micro-plastics, in particular to a method for separating sub-micron micro-plastics in water environment or marine products.
(II) technical background
The ocean is an important component upon which humans live and develop. There have been studies to find that over 5 trillion blocks of plastic in the world float on the sea, weighing 250000 tons. These plastic fragments are broken down into fine particles through physical, chemical and biological processes, and are currently becoming an important source of marine environmental pollution. Numerous studies have demonstrated that micro-plastics are threatening the survival of marine organisms and have been demonstrated by different marine species, which means that micro-plastics can accumulate in the marine food chain, affecting food safety and human health. At present, the pollution of micro-plastics in marine products is not regulated at home and abroad, but European countries have recognized the danger of the micro-plastics in food, and ecsafe seafoam firstly marks micro-and nano-plastic particles in the food as a potential food safety problem.
The toxicity of the marine products is directly influenced by the content of the micro-plastics in the marine products, and how to separate the micro-plastics from the marine products is a difficult point for establishing a quantitative analysis method of the micro-plastics. At present, the research on the isolation of microplastics is still in the initial stage and there is a lack of standardized methods for the isolation of microplastics in seafood. When the plastic fragments are larger than 1mm, they can be identified in a simple manner, i.e. visually with a microscope, but plastic particles smaller than 1mm cannot be identified in this way. Existing methods for micro-plastic chip separation typically involve the use of salt solution flotation or density separation. Although these methods have been commonly used, their collection efficiency is low and they are limited by the kind of plastic. Some researchers have succeeded in binding high density salts (NaI and ZnCl) using special equipment2) To separate micro-plastics from sediment, but the high density salts are expensive and this method is not suitable for samples with high organic content, such as waste water. On the basis of the above, the research finds that the defects can be overcome by enzyme digestion, peroxide digestion and acid digestion, but most of the post-treatments of the research are to filter and collect micro-plastics by using filter paper. These separation methods are only suitable for separating plastics larger than 40 microns, and can not completely separate plastics (such as submicron and nanometer) in the sample, and the fibers in the filter paper can interfere with the analysis of the sample.
Current analysis of microplastics has a size limit of about 20-40 μm, and the isolation of microplastics smaller than 20 μm is currently of greatest interest in the field of microplastics because they are more accessible to the body by uptake and other mechanisms. Therefore, the separation of submicron order micro plastic is a problem which needs to be solved urgently at present.
Disclosure of the invention
Aiming at the defects in the prior art, the invention provides a method for separating micro-plastics in water environment or marine products based on the combination of a double density separation method and a siphon method. The method can quickly and accurately separate and extract the submicron plastic particles in the sample, does not need suction filtration, centrifugation and drying, has simple operation and low cost, and provides technical support for subsequent accurate quantitative detection.
The technical scheme of the invention is as follows:
a method for separating submicron order micro-plastics, the method comprising the steps of:
(1) adding Sodium Dodecyl Sulfate (SDS) into a sample solution, ultrasonically dispersing uniformly, standing for 10-20 min, dividing the sample solution into a suspension layer, a solution layer and a precipitation layer from top to bottom, removing the intermediate solution layer through siphoning, and collecting the residue;
the dosage of the sodium dodecyl sulfate is 0.3-0.7 g/L based on the volume of the sample liquid;
when the middle solution layer is removed through siphoning, attention is paid to the fact that the middle solution layer cannot be removed completely, and a solution layer with the height of 0.2-0.3 cm is left between the suspension layer and the precipitation layer, so that particle loss of the suspension layer and the precipitation layer is prevented;
(2) adding 2-3 times of absolute ethyl alcohol by volume into the residue collected in the step (1), carrying out ultrasonic treatment (1-2 min), standing for 10-20 min, separating into two layers, removing the upper-layer solution through siphoning, and collecting the lower-layer substance;
(3) and (3) placing the lower-layer substance collected in the step (2) into a flat-bottomed vessel (such as a glass culture dish), adding 1-3 volume times of absolute ethyl alcohol, uniformly mixing, standing for 10-15 min, absorbing the upper-layer liquid after the two layers are separated, and naturally airing to separate the submicron-grade micro-plastic in the sample liquid.
Further, in the step (1), the sample solution is a water sample collected in a water environment or a digestion solution of marine products.
The digestion solution of the marine product is obtained by processing the following steps:
adding electronic grade HNO into dried marine product3Digesting at 45-50 ℃ for 10-20 min, cooling to room temperature (20-30 ℃), and adding analytical grade H2O2Continuously digesting for 10-20 min at 45-50 ℃, and then adding 10-15 volume times of water into the reaction liquid for dilution to obtain a digestion liquid;
the electronic grade HNO3The volume dosage of the composition is 16-23 mL/g based on the mass of the dried marine product;
the electronic grade HNO3And analytical grade H2O2The volume ratio of (A) to (B) is 4-8: 1, particularly preferably 8: 1.
further, in the step (1), after the intermediate solution layer sucked by the siphon is kept still (for 10-20 min), the intermediate solution layer is divided into 3 layers from top to bottom, the intermediate layer is continuously removed by the siphon, and the residual substances and the collected substances in the previous time are combined.
Further, in the step (2), after standing still (10-20 min), the upper layer solution sucked by the siphon is divided into two layers, the upper layer solution is continuously removed by the siphon, and the lower layer substance is combined with the substance collected in the previous time.
Compared with the prior art, the invention has the beneficial effects that:
1. the method is simple to operate, the separation liquid can be recycled, filter paper filtration, centrifugation and drying in an oven are not needed, and submicron plastic particles in marine products can be extracted quickly;
2. the separating liquid has less consumption, low cost and easy obtaining, wherein the SDS can also be used for removing trace organic matters adsorbed on the surface of the micro-plastic;
3. compared with the conventional filter paper filtration and separation, the invention has the advantages that the plastic particle loss caused in the separation process is less, the plastic particle loss can be used for separating micro plastic with the particle size of less than 20 micrometers, and the interference on the subsequent identification and quantitative analysis can not be caused;
4. the invention can be used to separate submicron order micro-plastics in seafood and water samples, which can provide technical support for food safety work.
(IV) description of the drawings
FIG. 1: the siphon separation device of the invention is shown schematically;
FIG. 2: the invention discloses a separation process flow chart;
FIG. 3: example 1 flow chart of the process for separating microplastic (mixed system of PVC, PS, PE) from marine shrimp.
(V) detailed description of the preferred embodiments
The present invention is further illustrated by the following specific examples, but the scope of the invention is not limited thereto.
HNO used in the following examples3(electronic grade, 70%) from Shanghai Aladdin reagent; h2O2(AR，30wt.％in H2O2) Purchased from Hangzhou institute general and herbal and textbook equipment Co.
Example 1 separation of microplastics from Marine product shrimp
(1) Weighing 0.336g of freeze-dried prawns into a beaker, adding 0.0286g of micro-plastic standard PVC (polyvinyl chloride) with the particle size of less than 20 mu m into the prawns as an internal standard, adding 8mL of nitric acid into the beaker, placing the beaker on a heating plate, heating at 50 ℃ for 20min, taking down the beaker, cooling, adding 1mL of hydrogen peroxide, digesting for 15min under the same condition, taking down the beaker after the sample is completely digested, cooling to room temperature, and adding 150mL of water into a reaction solution for dilution to obtain a digestion solution.
(2) Adding 0.05g of sodium dodecyl sulfate into the obtained digestion solution (159mL), ultrasonically dispersing uniformly (70Hz for 1-2min), standing for 15min, dividing the solution into a suspension layer, a solution layer and a precipitation layer from top to bottom, removing the intermediate solution layer through siphoning, and collecting the rest; standing the intermediate solution layer for 15min, dividing into 3 layers from top to bottom, removing the intermediate layer by siphoning, and mixing the residue with the residue collected from the previous time;
when removing the intermediate solution layer by siphoning, attention was paid to the fact that the intermediate solution layer could not be removed completely, and a solution layer with a height of 0.2cm remained between the suspension layer and the precipitation layer to prevent the loss of particles from the suspension layer and the precipitation layer.
(3) Adding 50mL of anhydrous ethanol into the collected residue (25mL), performing ultrasonic treatment (70Hz for 2min), standing for 15min, separating into two layers, removing the upper layer solution by siphoning, and collecting the lower layer substance; taking the upper layer solution sucked by siphon, standing for 15min, dividing into two layers, removing the upper layer solution by siphon, and combining the lower layer material with the previous collected material.
(4) And (3) placing the collected lower-layer substances (10mL) in a glass culture dish, adding 10mL of absolute ethyl alcohol, uniformly mixing, standing for 15min, sucking the upper-layer liquid after the lower-layer substances are separated into two layers, and naturally airing to separate the upper-layer liquid into submicron-grade micro-plastics in the prawns.
Single standard, mixed standard spiking experiments were performed on the stock samples following the procedure of steps (1) to (4) above to verify process feasibility, with the results shown in tables 1 and 2:
table 1: labeling result of single standard substance in marine product prawn
Table 2: labeling result of mixed standard substance in marine product prawn
Example 2 separation of microplastics in aqueous environments
A water sample of a Hangzhou Shanggan river in Zhejiang is used as a test sample, and the separation process is as follows:
(1) taking 150mL of water sample, adding 0.031g of a micro-plastic standard PVC with the particle size of less than 20 mu m into the water sample as an internal standard, then adding 0.04g of sodium dodecyl sulfate, ultrasonically dispersing uniformly (70Hz for 1-2min), standing for 15min, then dividing into 3 layers from top to bottom, namely a suspension layer, a solution layer and a precipitation layer, removing the middle solution layer through siphoning, and collecting the rest; standing the intermediate solution layer for 15min, dividing into 3 layers from top to bottom, removing the intermediate layer by siphoning, and mixing the residue with the residue collected from the previous time;
when removing the intermediate solution layer by siphoning, attention was paid to the fact that the intermediate solution layer could not be removed completely, and a solution layer with a height of 0.2cm remained between the suspension layer and the precipitation layer to prevent the loss of particles from the suspension layer and the precipitation layer.
(2) Adding 50mL of anhydrous ethanol into the collected residue (25mL), performing ultrasonic treatment (70Hz for 2min), standing for 15min, separating into two layers, removing the upper layer solution by siphoning, and collecting the lower layer substance; taking the upper layer solution sucked by siphon, standing for 15min, dividing into two layers, removing the upper layer solution by siphon, and combining the lower layer material with the previous collected material.
(3) And (3) placing the collected lower-layer substance (10mL) in a glass culture dish, adding 10mL of absolute ethyl alcohol, uniformly mixing, standing for 15min, sucking the upper-layer liquid after the lower-layer substance is divided into two layers, and naturally airing to separate the upper-layer liquid into submicron-grade micro-plastics in a water sample.
A single standard spiking experiment was performed on the stock samples following the procedure of steps (1) - (3) above to verify process feasibility, with the results shown in table 3:
table 3: single standard substance adding result in water sample
| Name of micro-plastic | Mass of standard substance (g) | Mass after separation (g) | Recovery (%) |
| PVC | 0.031 | 0.0295 | 95.16％ |
| PS | 0.0322 | 0.0308 | 95.65％ |
| PE | 0.018 | 0.017 | 94.44％ |
Claims (7)
1. A method for separating submicron order micro-plastic, comprising the steps of:
(1) adding sodium dodecyl sulfate into a sample solution, ultrasonically dispersing uniformly, standing, then dividing into a suspension layer, a solution layer and a precipitation layer from top to bottom, removing the middle solution layer through siphoning, and collecting the residue;
the dosage of the sodium dodecyl sulfate is 0.3-0.7 g/L based on the volume of the sample liquid;
(2) adding 2-3 times by volume of absolute ethyl alcohol into the residue collected in the step (1), standing after ultrasonic treatment to separate the mixture into two layers, removing the upper layer solution through siphoning, and collecting the lower layer substance;
(3) and (3) placing the lower-layer substance collected in the step (2) into a flat bottom vessel, adding 1-3 volume times of absolute ethyl alcohol, uniformly mixing, standing until the lower-layer substance is divided into two layers, absorbing the upper-layer liquid, and naturally airing to separate the upper-layer liquid into submicron-grade micro plastic in the sample liquid.
2. The method for separating submicron order microplastic according to claim 1, wherein in step (1), the sample solution is a water sample collected in an aqueous environment or a digestion solution of marine products.
3. The method for separating submicron order microplastic according to claim 2, wherein the digestion solution of marine products is obtained by the following steps:
adding electronic grade HNO into dried marine product3Digesting at 45-50 ℃ for 10-20 min, cooling to room temperature, and adding analytical grade H2O2Continuously digesting for 10-20 min at 45-50 ℃, and then adding 10-15 volume times of water into the reaction liquid for dilution to obtain a digestion liquid;
the electronic grade HNO3The volume dosage of the composition is 16-23 mL/g based on the mass of the dried marine product;
the electronic grade HNO3And analytical grade H2O2The volume ratio of (A) to (B) is 4-8: 1.
4. the method for separating submicron-sized micro-plastic according to claim 3, wherein the electronic-grade HNO3And analytical grade H2O2Is 8: 1.
5. the method for separating submicron order microplastic according to claim 1, wherein in step (1), when the intermediate solution layer is removed by siphoning, a solution layer with a height of 0.2-0.3 cm is left between the suspension layer and the precipitation layer.
6. The method for separating submicron order microplastic according to claim 1, wherein in step (1), the layer of the intermediate solution sucked by the siphon is allowed to stand and then is divided into 3 layers from top to bottom, and the removal of the intermediate layer by the siphon is continued to combine the remainder with the previous collection.
7. The method for separating submicron order microplastic according to claim 1, wherein in step (2), the upper solution sucked by the siphon is allowed to stand and is then separated into two layers, and the upper solution is further removed by siphon to combine the lower layer with the previous collected material.
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| CN110186863A (en) * | 2019-05-08 | 2019-08-30 | 浙江工业大学 | The detection method of the micro- plastic degradation situation in mulch source in a kind of pair of soil |
| ES2796998A1 (en) | 2019-05-31 | 2020-11-30 | Univ Madrid Autonoma | Procedure for the separation of microplastics from aqueous matrices (Machine-translation by Google Translate, not legally binding) |
| CN111504741B (en) * | 2020-04-22 | 2023-04-18 | 山东师范大学 | Method for detecting micro-plastics in fish body |
| CN112525635B (en) * | 2020-11-20 | 2022-01-04 | 哈尔滨工业大学（深圳） | Method for extracting micro-plastic |
| CN112903349A (en) * | 2021-01-19 | 2021-06-04 | 河南省科学院高新技术研究中心 | Method for extracting and detecting micro-plastics in urban river sediment |
| CN113447466B (en) * | 2021-05-24 | 2022-11-01 | 暨南大学 | Method for detecting abundance of micro-plastics in aquatic organisms |
| JP7299450B2 (en) * | 2021-06-14 | 2023-06-28 | 東レテクノ株式会社 | Method for separating synthetic resin pieces from water containing synthetic resin pieces |
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