CN108181154A - The detection method of micro- plastics in a kind of organism - Google Patents
The detection method of micro- plastics in a kind of organism Download PDFInfo
- Publication number
- CN108181154A CN108181154A CN201711178353.0A CN201711178353A CN108181154A CN 108181154 A CN108181154 A CN 108181154A CN 201711178353 A CN201711178353 A CN 201711178353A CN 108181154 A CN108181154 A CN 108181154A
- Authority
- CN
- China
- Prior art keywords
- micro
- plastics
- organism
- detection
- water
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 229920003023 plastic Polymers 0.000 title claims abstract description 143
- 239000004033 plastic Substances 0.000 title claims abstract description 143
- 238000001514 detection method Methods 0.000 title claims abstract description 22
- 238000000034 method Methods 0.000 claims abstract description 36
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 19
- GRYLNZFGIOXLOG-UHFFFAOYSA-N Nitric acid Chemical compound O[N+]([O-])=O GRYLNZFGIOXLOG-UHFFFAOYSA-N 0.000 claims abstract description 15
- 238000005238 degreasing Methods 0.000 claims abstract description 15
- 229910017604 nitric acid Inorganic materials 0.000 claims abstract description 14
- 239000004094 surface-active agent Substances 0.000 claims abstract description 7
- 241000237502 Ostreidae Species 0.000 claims description 40
- 235000020636 oyster Nutrition 0.000 claims description 40
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 39
- 238000004043 dyeing Methods 0.000 claims description 17
- -1 Oxygen alkane Chemical class 0.000 claims description 12
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 claims description 11
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical class [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 10
- 238000000967 suction filtration Methods 0.000 claims description 10
- GNBHRKFJIUUOQI-UHFFFAOYSA-N fluorescein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 GNBHRKFJIUUOQI-UHFFFAOYSA-N 0.000 claims description 8
- YUYCVXFAYWRXLS-UHFFFAOYSA-N trimethoxysilane Chemical compound CO[SiH](OC)OC YUYCVXFAYWRXLS-UHFFFAOYSA-N 0.000 claims description 8
- RCEAADKTGXTDOA-UHFFFAOYSA-N OS(O)(=O)=O.CCCCCCCCCCCC[Na] Chemical group OS(O)(=O)=O.CCCCCCCCCCCC[Na] RCEAADKTGXTDOA-UHFFFAOYSA-N 0.000 claims description 6
- 238000004140 cleaning Methods 0.000 claims description 6
- 239000000975 dye Substances 0.000 claims description 5
- 238000002329 infrared spectrum Methods 0.000 claims description 5
- 239000000463 material Substances 0.000 claims description 5
- 239000002245 particle Substances 0.000 claims description 5
- 229920013822 aminosilicone Polymers 0.000 claims description 4
- IDGUHHHQCWSQLU-UHFFFAOYSA-N ethanol;hydrate Chemical compound O.CCO IDGUHHHQCWSQLU-UHFFFAOYSA-N 0.000 claims description 3
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 2
- 239000001110 calcium chloride Substances 0.000 claims description 2
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 2
- 239000000725 suspension Substances 0.000 claims description 2
- 229910052760 oxygen Inorganic materials 0.000 claims 1
- 239000001301 oxygen Substances 0.000 claims 1
- 230000007613 environmental effect Effects 0.000 abstract description 3
- 238000009826 distribution Methods 0.000 abstract description 2
- 238000001727 in vivo Methods 0.000 abstract description 2
- 238000012544 monitoring process Methods 0.000 abstract description 2
- 238000002474 experimental method Methods 0.000 description 55
- 239000000243 solution Substances 0.000 description 31
- 239000000523 sample Substances 0.000 description 19
- 239000004743 Polypropylene Substances 0.000 description 18
- 239000007788 liquid Substances 0.000 description 18
- 230000029087 digestion Effects 0.000 description 14
- 229910021642 ultra pure water Inorganic materials 0.000 description 13
- 239000012498 ultrapure water Substances 0.000 description 13
- 239000003643 water by type Substances 0.000 description 13
- 238000001157 Fourier transform infrared spectrum Methods 0.000 description 10
- 238000010438 heat treatment Methods 0.000 description 10
- 239000004800 polyvinyl chloride Substances 0.000 description 10
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 9
- 239000004519 grease Substances 0.000 description 9
- 239000012528 membrane Substances 0.000 description 9
- 229920001155 polypropylene Polymers 0.000 description 9
- 235000019483 Peanut oil Nutrition 0.000 description 8
- 239000003925 fat Substances 0.000 description 8
- 239000000312 peanut oil Substances 0.000 description 8
- 238000000605 extraction Methods 0.000 description 7
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 6
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 description 6
- 238000002604 ultrasonography Methods 0.000 description 6
- 238000002203 pretreatment Methods 0.000 description 5
- 239000000126 substance Substances 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 4
- 239000003599 detergent Substances 0.000 description 4
- 238000005516 engineering process Methods 0.000 description 4
- 238000012986 modification Methods 0.000 description 4
- 230000004048 modification Effects 0.000 description 4
- 235000019834 papain Nutrition 0.000 description 4
- IOLCXVTUBQKXJR-UHFFFAOYSA-M potassium bromide Chemical compound [K+].[Br-] IOLCXVTUBQKXJR-UHFFFAOYSA-M 0.000 description 4
- 239000007787 solid Substances 0.000 description 4
- 239000004365 Protease Substances 0.000 description 3
- 230000008859 change Effects 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 235000019441 ethanol Nutrition 0.000 description 3
- 238000000799 fluorescence microscopy Methods 0.000 description 3
- 239000006260 foam Substances 0.000 description 3
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 3
- 238000001000 micrograph Methods 0.000 description 3
- 229920001343 polytetrafluoroethylene Polymers 0.000 description 3
- 239000004810 polytetrafluoroethylene Substances 0.000 description 3
- 238000002444 silanisation Methods 0.000 description 3
- 239000011780 sodium chloride Substances 0.000 description 3
- 235000002639 sodium chloride Nutrition 0.000 description 3
- 108090000790 Enzymes Proteins 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 2
- 108090000526 Papain Proteins 0.000 description 2
- 239000004698 Polyethylene Substances 0.000 description 2
- 230000000903 blocking effect Effects 0.000 description 2
- 238000012512 characterization method Methods 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 230000006835 compression Effects 0.000 description 2
- 238000007906 compression Methods 0.000 description 2
- 239000004035 construction material Substances 0.000 description 2
- 238000001816 cooling Methods 0.000 description 2
- 229940088598 enzyme Drugs 0.000 description 2
- 239000011521 glass Substances 0.000 description 2
- 239000012535 impurity Substances 0.000 description 2
- 239000005416 organic matter Substances 0.000 description 2
- 230000010355 oscillation Effects 0.000 description 2
- 229940055729 papain Drugs 0.000 description 2
- 239000011236 particulate material Substances 0.000 description 2
- 229920000573 polyethylene Polymers 0.000 description 2
- 229920000915 polyvinyl chloride Polymers 0.000 description 2
- 239000003755 preservative agent Substances 0.000 description 2
- 230000002335 preservative effect Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 230000002000 scavenging effect Effects 0.000 description 2
- 239000002002 slurry Substances 0.000 description 2
- 239000011734 sodium Substances 0.000 description 2
- 229910052708 sodium Inorganic materials 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 231100000027 toxicology Toxicity 0.000 description 2
- 230000000007 visual effect Effects 0.000 description 2
- 241001436679 Adama Species 0.000 description 1
- RZYKUPXRYIOEME-UHFFFAOYSA-N CCCCCCCCCCCC[S] Chemical compound CCCCCCCCCCCC[S] RZYKUPXRYIOEME-UHFFFAOYSA-N 0.000 description 1
- 102000057297 Pepsin A Human genes 0.000 description 1
- 108090000284 Pepsin A Proteins 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 239000004793 Polystyrene Substances 0.000 description 1
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 1
- BQCADISMDOOEFD-UHFFFAOYSA-N Silver Chemical compound [Ag] BQCADISMDOOEFD-UHFFFAOYSA-N 0.000 description 1
- 230000005856 abnormality Effects 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 238000000184 acid digestion Methods 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 235000013361 beverage Nutrition 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000005660 chlorination reaction Methods 0.000 description 1
- 238000004040 coloring Methods 0.000 description 1
- 238000009833 condensation Methods 0.000 description 1
- 230000005494 condensation Effects 0.000 description 1
- 239000000356 contaminant Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000001493 electron microscopy Methods 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000010419 fine particle Substances 0.000 description 1
- 125000000524 functional group Chemical group 0.000 description 1
- 230000004077 genetic alteration Effects 0.000 description 1
- 230000002070 germicidal effect Effects 0.000 description 1
- 238000000227 grinding Methods 0.000 description 1
- 229940116364 hard fat Drugs 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 238000009533 lab test Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 235000010755 mineral Nutrition 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000000465 moulding Methods 0.000 description 1
- 238000011017 operating method Methods 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 239000005022 packaging material Substances 0.000 description 1
- 229940111202 pepsin Drugs 0.000 description 1
- 229920001084 poly(chloroprene) Polymers 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- QQONPFPTGQHPMA-UHFFFAOYSA-N propylene Natural products CC=C QQONPFPTGQHPMA-UHFFFAOYSA-N 0.000 description 1
- 125000004805 propylene group Chemical group [H]C([H])([H])C([H])([*:1])C([H])([H])[*:2] 0.000 description 1
- 235000019419 proteases Nutrition 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 239000004576 sand Substances 0.000 description 1
- 238000007127 saponification reaction Methods 0.000 description 1
- 239000013535 sea water Substances 0.000 description 1
- 235000014102 seafood Nutrition 0.000 description 1
- 230000035939 shock Effects 0.000 description 1
- 229910052709 silver Inorganic materials 0.000 description 1
- 239000004332 silver Substances 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 239000000344 soap Substances 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 238000013517 stratification Methods 0.000 description 1
- 238000009941 weaving Methods 0.000 description 1
Classifications
-
- 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
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
-
- 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
- G01N1/34—Purifying; Cleaning
-
- 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
- G01N1/40—Concentrating samples
- G01N1/4044—Concentrating samples by chemical techniques; Digestion; Chemical decomposition
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Optical investigation techniques, e.g. flow cytometry
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/35—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using infrared light
- G01N21/3563—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using infrared light for analysing solids; Preparation of samples therefor
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N23/00—Investigating or analysing materials by the use of wave or particle radiation, e.g. X-rays or neutrons, not covered by groups G01N3/00 – G01N17/00, G01N21/00 or G01N22/00
- G01N23/22—Investigating or analysing materials by the use of wave or particle radiation, e.g. X-rays or neutrons, not covered by groups G01N3/00 – G01N17/00, G01N21/00 or G01N22/00 by measuring secondary emission from the material
- G01N23/2202—Preparing specimens therefor
-
- 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
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
- G01N2001/302—Stain compositions
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Optical investigation techniques, e.g. flow cytometry
- G01N2015/1486—Counting the particles
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/35—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using infrared light
- G01N21/3563—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using infrared light for analysing solids; Preparation of samples therefor
- G01N2021/3572—Preparation of samples, e.g. salt matrices
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/35—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using infrared light
- G01N2021/3595—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using infrared light using FTIR
Landscapes
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- General Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Dispersion Chemistry (AREA)
- Sampling And Sample Adjustment (AREA)
Abstract
The present invention relates to it is a kind of detect Organism Samples in micro- plastics method, including：S110, resolution：Organism Samples with a concentration of 50 70% nitric acid are impregnated 6 18 hours at room temperature, be heated to 60 95 DEG C and are maintained 14 hours, the Organism Samples cleared up；S120, degreasing：The Organism Samples of resolution obtained in step S110 are rinsed and filtered with 60 95 DEG C of surfactant solution while hot, obtain micro- plastics of degreasing；Micro- plastics of the degreasing obtained in step S120 are rinsed and are filtered, the micro- plastics cleaned with 60 95 DEG C of water by S130.The method of the present invention quickly and accurately can carry out qualitative and quantitative detection to the micro- plastics entered in organism, realize the purpose for detecting the enrichment in vivo of micro- plastics and distribution, and the tool of practicality is provided for environmental monitoring.
Description
Technical field
The invention belongs to environmental contaminants detection field, more particularly, to a kind of detection method of micro- plastics in organism,
The method of micro- plastics in particularly a kind of method detection organism counted using fluorescence microscope.
Background technology
Plastics since its is at low cost, stablize by chemical property, the features such as shock resistance is good, is largely made in daily life
With, but because it is difficult to degrade, environmental pollution is exacerbated, ocean plastic pollution situation is especially prominent.Plastic pollution object in seawater
Molecule is gradually degraded into factors such as light, heat, biologies, referred to as micro- plastics.Once micro- plastic pollution raising in water, just
Toxicology can be caused to react marine organisms, such as induce genetic aberrations, toxicology occur in reproduction abnormality, hormone disturbance or tissue
Variation etc..The ingredient and content of micro- plastics in oyster are studied, its pollution mechanism can be not only illustrated, more can be seafood security
It provides safeguard.
Therefore, there is still a need for the method for detecting micro- plastics in organism that exploitation is new.
Invention content
In order to solve above-mentioned the problems of the prior art, the present invention provides following technical schemes.
One aspect of the present invention provides a kind of method for detecting micro- plastics in Organism Samples, including：
S100, the step of micro- plastics are extracted from Organism Samples, including：
S110, resolution：Organism Samples with the nitric acid of a concentration of 50-70% are impregnated 6-18 hours at room temperature, are heated
To 60-95 DEG C and maintain 1-4 hours, the Organism Samples cleared up；
S120, degreasing：The Organism Samples of resolution obtained in step S110 are used to 60-95 DEG C of surfactant while hot
Solution is rinsed and is filtered, and obtains micro- plastics of degreasing；
The water of 60-95 DEG C of micro- plastics of the degreasing obtained in step S120 is rinsed and filtered, cleaned by S130
Micro- plastics.
Further, step S100 is further included：Micro- plastics of obtained cleaning are added to the water, is then ultrasonically treated, obtains
To micro- plastics suspension.
Further, the suction filtration is the suction filtration carried out with miillpore filter.
Further, step S100 is further included：
S140, classification：Micro- plastics are pressed into density classification with the different solution of density；Preferably, the solution is selected from density
For 1g/cm3Water, density 1.19g/cm3Saturated sodium-chloride water solution, density 0.91g/cm358.4% ethyl alcohol it is water-soluble
Liquid, density 0.925g/cm355.4% ethanol water and density be 1.27g/cm327.5% calcium chloride water.
Further, the surfactant is lauryl sodium sulfate.
Further, the method for detecting micro- plastics in organism, further includes：
S200 dyes micro- plastics of the cleaning obtained in step S100, including：With amino silicone and micro- plastics
Micro- plastics to obtain having amino are contacted, are then contaminated with fluorescein isothiocynate and micro- plastic contacts with amino
Micro- plastics of color.
Further, in step s 200, the amino silicone is (3- aminopropyls) trimethoxy silane, according to 19:1
Ratio methanol and (3- aminopropyls) trimethoxy silane are added in micro- plastics to obtain micro- plastics with amino.
Further, the method for detecting micro- plastics in organism, further includes：
S300 with micro- plastics of the dyeing obtained in microscope observation of steps S200, and counts the particle of micro- plastics
Number.
Further, the method for detecting micro- plastics in organism, further includes：
Micro- plastics of the cleaning obtained in step S100 are carried out infrared spectrum detection or electron microscope characterize by S400.
It is dyed using silyation technigue by exploring and optimizing micro- plastics extraction process, secondly by the present invention and fluorimeter
Number, is finally characterized and is analyzed to plastics kind using FTIR spectrum and electron microscope (SEM).Particularly, lead to
It crosses and micro- plastics is extracted from oyster, and Optimized Extraction Process, while using silyation technigue to micro- plastics combination fluorescence of extraction
It is dyed, finally by FTIR spectrum (FT-IR) and electron microscope (SEM), observes micro- plastics kind and pattern
Feature.Research has shown that：Really containing micro- plastics in the oyster of Zhanjiang sea area, best extraction process is carried out with 25mL concentrated nitric acids
Digestion, lauryl sodium sulfate remove fat as surfactant；Optimal colouring method is with methanol and (3- aminopropyls)
Trimethoxy silane modifies amino to micro- frosting, and amino combines with fluorescein isothiocynate (FITC) and generates fluorescence, passes through
Fluorescence microscope counts, and sample includes 67368/g.Micro- plastics are analyzed through electron microscope (SEM) and FTIR spectrum
Sample may contain polypropylene (PP) or polyvinyl chloride (PVC).
The method of the present invention quickly and accurately can carry out qualitative and quantitative detection to the micro- plastics entered in organism,
The purpose for detecting the enrichment in vivo of micro- plastics and distribution is realized, the tool of practicality is provided for environmental monitoring.
Description of the drawings
Fig. 1 is micro- plastics light field micrograph (4x) in oyster, wherein (a), (b) and (c) is the image in different visual fields；
Fig. 2 is the light field micrograph (4x) of the fritter plastics shredded on polybag in dyeing of plastics preliminary experiment；
Fig. 3 is that form of the plastics of above FITC under 4 times of details in a play not acted out on stage, but told through dialogues of microscope is modified in dyeing of plastics preliminary experiment, wherein (a),
(b) and (c) be modify FITC plastics fluorescence microscopy figure (4x)；
Fig. 4 is the form of micro- dyeing of plastics being clearly observable in micro- dyeing of plastics preliminary experiment, wherein (a), (b) and (c)
It is the fluorescence microscopy figure (4x) of micro- plastics in oyster；
Fig. 5 is the experimental result of micro- dyeing of plastics, wherein the fluorescence that (a), (b) and (c) is micro- plastics in oyster counts
Micrograph (4x)；
Fig. 6 is the infrared spectrogram of micro- plastics in oyster；
Infrared spectrograms of the Fig. 7 for polypropylene in the prior art (figure is left) and polyvinyl chloride (figure is right)；
Fig. 8 is the image of micro- plastic sample under an electron microscope in oyster, wherein (a), (b) and (c) is in oyster
Micro- plastic electronic microscope figure.
Specific embodiment
It is described below and illustrates the exemplary implementation of the present invention.It is discussed below described in order to clear and accurate
Exemplary implementation can include preferred step, method and feature, and those of ordinary skill in the art will be appreciated that, this
A little preferred step, method and features are not the necessary conditions being within the scope of the present invention.
Used experimental method is conventional method unless otherwise specified in the following embodiments.In following embodiments
Used in material, reagent etc. be commercially available unless otherwise specified.
Embodiment 1：The detection of micro- plastics in oyster
1.1 the pre-treatment of oyster
(1) multiple fishings from the oyster of Zhanjiang sea area from refrigerator are taken out and thawed, remove shell, washed with water clean.
(2) oyster cleaned up on electronic balance is weighed, and records weight respectively.
The exploration of the experiment condition of 1.2 digestion
Following several method is taken to the resolution of oyster：Enzyme resolution method, nitric acid and the resolution of 30% hydrogen peroxide, nitric acid disappear
Solution, saturated sodium-chloride top and bottom process and ethyl alcohol stratification method.
1.2.1 enzyme resolution method
(1) 5 oysters (114g) are taken out from refrigerator, after 3.1 step pre-treatments, with juice extractor, (JYL-C52V is more
Energy juice extractor, JiuYang Co., Ltd；Similarly hereinafter) smash it is slurry, add in 500mL water, 5g papains (papain,
Guangzhou Qi Yun Bioisystech Co., Ltd；It after similarly hereinafter), stands overnight, whether observation protease can decompose oyster.
(2) oyster (11.52g) is taken, after 3.1 step pre-treatments, slurry, addition 50mL water is smashed with juice extractor
Afterwards, 30min is boiled, adds 0.9g papains, and stood overnight in 50 DEG C of water-baths.
Method according to experiment (1) is tested, and after finding solution left standstill overnight, muddy green is still presented, simultaneously
It can be observed have block objects, can be initially believed that the method resolution is unsuccessful.Experiment (1) is optimized, oyster heating,
Make its protein denaturation, remove most of organic matter, while when being cooled to about 50 DEG C, add in papain, and in water-bath
It is kept for 50 DEG C in pot to stand overnight, allows the remaining organic matter of pepsin digestion.After overnight, solution is muddy in baby pink shape
It is turbid, therefore add in 58% ethyl alcohol and enable its layering, but effect still unobvious, without the liquid layer of clear.
1.2.2 nitric acid resolution method
The oyster of pre-treatment is taken, with ultra-pure water rinse one time, is put into kjeldahl flask, carries out the experiment in table 1.
Table 1：Nitric acid clears up serial experiment
The combination of 18mL 69%HNO3+6mL 30%H2O2,69% nitric acid and 30% hydrogen peroxide 3 are used first:1
Ratio is cleared up, and carries out 1.2.2 experiments 1,2, and the heating of 10 minutes is carried out to experiment 1, then quiet using 12 hours to experiment 2
Overnight method is put, carries out comparison reference.It was found that the heating of 10 minutes can clear up whole oyster in experiment 1, but deposit
In a large amount of foams and similar grease-like substance, bottom fine particle is more also deposited in, preliminary judgement is fine sand, therefore,
A large amount of foam with and similar grease-like substance can be determined that the experiment fails to clear up completely substantially；12 hours are stood in experiment 2
Afterwards, still fail to fall the resolution of whole oyster, the heating of 5 minutes is then carried out to it, oyster is just cleared up for 1 state of experiment.
In conclusion not yet determine whether digestion solution is effective at present, but what is certain is that, if heating and heating time
Length by the degree of the resolution of influence, for this purpose, when test in next step, will keep heating, and control heating time, simultaneously
Change time of repose, probe into influence degree of the time of repose to resolution.
Next, directly using 69% nitric acid as digestion solution, and stand overnight before heating, be heated to boil condition simultaneously
It is kept for 2 hours, then devises the serial experiment 3,4,5,6 in table 1.Experimental result is shown in Table 2.
Table 2：Nitric acid clears up serial experiment result
As a result it can be found that there are a large amount of bulk particulate materials, concentration in experiment 3 to float on digestion solution upper strata, liquid is in
Light yellow shape is tentatively judged as that fat condenses into bulk, is unfavorable for extracting in next step, meanwhile, experiment 5, experiment 6 occur identical
Situation, it may be possible to it is too long due to standing digestion time, cause fat cooling condensation blocking, only test 4, the grease on surface is to be in
It is liquid, therefore, can tentatively judgment experiment 3, experiment 5 and test 6 condition it is infeasible, the reason is that a large amount of fat solid is unfavorable
It is extracted in next step, the micro- plastics that may include hard fat if direct removal remove together, and test the fat in 4
In liquid, it is only necessary to by itself and water remove together can, method most viable at present is using Suction filtration device, chooses appropriate bore
The filter membrane of diameter, you can stay in micro- plastics on filter membrane, remaining impurity will together be filtered with water.
1.2.3 degreasing is tested
The oyster of pre-treatment is taken, with ultra-pure water rinse oyster, is put into kjeldahl flask, according to the method for experiment 4 in 1.2.2
After processing, experiment 9,10 is carried out according to following scheme, wherein experiment 7,8 is the preliminary experiment of degreasing, is made with reference to effect.
3 degreasing of table is tested
Pass through the experiment 4 of 1.2.2, it is thus understood that there are a large amount of liquid fats on digestion solution surface, select 0.22 μm of miillpore filter (on
Haixing County Asia scavenging material factory) it is filtered, it is found that liquid fat can not penetrate filter membrane.Micro- plastics aperture of this experimental study has
Less than 5mm's.Therefore, 9 schemes of experiment are carried out, by adding in 1mol NaOH during filtering, enable grease and highly basic that soap occurs
Change reaction, and use a large amount of ultrapure waters, so as to eliminate the influence of grease, but actual effect is that saponification has occurred in grease and highly basic
Reaction, can not make grease become small molecule and be filtered, and filter membrane surface still remains a large amount of greases, block Suction filtration device, so
Go out to remove the scheme of grease and infeasible with NaOH.
In the case where NaOH schemes are infeasible, it is contemplated that the daily lifes such as liquid detergent, hand cleanser are added in except grease detergent, but these
Detergent may contain micro- plastics originally, under the premise of impurity is not introduced, the surface similar to these detergents be selected to live
Property agent, therefore, select lauryl sodium sulfate (chemical pure, Guangzhou brilliance Science and Technology Co., Ltd.), and carried out with peanut oil
Preliminary experiment.
1.2.3 experiment 7,8 is the preliminary experiment of peanut oil, and lauryl sodium sulfate is added without in experiment 7, will be directly added dropwise
The water of few drops of peanut oil is filtered.It was found that there are a large amount of peanut oil on miillpore filter, it is impossible to through miillpore filter, and block
Device is filtered, even if adding in ultra-pure water can not continue to filter；Experiment 8 adds in sodium dodecyl sulfate solution i.e. after suction filtration, micro-
Peanut oil on the filter membrane of hole has significantly through filter membrane, and receiver has a large amount of foams to occur, and adding in ultra-pure water can be quickly through micro-
There is not the clogging in experiment 7 in hole filter membrane, and therefore, sodium dodecyl sulfate solution can effectively make liquid in preliminary experiment
Body grease penetrates 0.22 μm of miillpore filter.
According to the feasibility of peanut oil preliminary experiment, experiment 10 is designed, after the completion of oyster is cleared up, adds in 80 DEG C of ultra-pure waters
10 times of dilution, and filter while hot, while add in 80 DEG C of sodium dodecyl sulfate solutions, penetrate the liquid fat on miillpore filter
Filter membrane, finally with a large amount of 80 DEG C of ultrapure waters, Germicidal efficacy arrives, and has a small amount of light yellow particulate material on miillpore filter, can
Be judged as cooling grease solid small particles, and will not blocking microporous filter membrane, ultra-pure water can fast strikethrough miillpore filter, can recognize
Concept feasible thus.
Reagent preparation：
(1) sodium dodecyl sulfate solution：5g lauryl sodium sulfate is taken to add in 50mL ultra-pure waters.
(2) 1molNaOH solution：2gNaOH is taken to be dissolved in 50mL ultra-pure waters.
Experiment 7：A beaker is taken, few drops of peanut oil are added dropwise in 50mL ultra-pure waters, are filtered with 0.22 μm of miillpore filter.
Experiment 8：It takes a beaker, few drops of peanut oil is added dropwise in 50mL ultra-pure waters, are filtered with 0.22 μm of miillpore filter, then
Add in sodium dodecyl sulfate solution degreasing.
Experiment 9：The digestion solution of 1.2.2 is taken, is filtered with 0.22 μm of miillpore filter, it is molten that 1molNaOH is added in after suction filtration
Liquid.
Experiment 10：The digestion solution of 1.2.2 is taken, is filtered with 0.22 μm of miillpore filter, dodecyl sulphur is added in after suction filtration
Acid sodium solution degreasing.
1.2.4 microexamination
Will 1.2.2 test 4 in filter the miillpore filter of completion, be put into the beaker equipped with 20mL ultra-pure waters, and with ultrasonic
30 seconds, 20mL saturation NaCl solutions are then added in, 12 hours is stood, waits to be layered.After 12 hours, the 20 μ L upper stratas for taking solution are clear
Liquid is added drop-wise on clean glass slide, with microscope (Olympus IX73 inverted microscopes, Olympus Co., Ltd) bright
It is observed off field.
In order to which micro- plastics is made preferably to be observed, need to assemble the micro- plastics in part using densimetry, according to 4 plastics of table
Sink-float phenomenon, saturation NaCl solution is selected to be screened into line density, reason is that it is safer, in addition will not be completed with digestion
Possible remaining nitric acid reacts.
Table 4：Sink-float phenomenon table of the common plastics in different densities solution
Digestion is completed according to the operating method of the experiment 4 of 1.2.2 and is layered using saturation NaCl solution, draws upper strata
Solution is observed under 4 power microscopes, and the results are shown in Figure 1, wherein figure (a), (b) and (c) is the figure under different visual fields.It can
To see, some block objects are in regular transparence, and large number of, some volumes are relatively large, and plastic features are apparent, can be first
Step is judged as micro- plastics.
The quantitative measurement of micro- plastics in 1.3 oysters
1.3.1 micro- dyeing of plastics experiment
By preliminary micro- plastic sight, in order to further definitely prove the presence of micro- plastics, and it is counted, profit
Modification amino is carried out to its surface with silyation technigue, the amino in modification is made to be combined with fluorescein, is seen under fluorescence microscope
Examine and count, for this purpose, design plastics silanization preliminary experiment, it was demonstrated that plastics silanization it is feasible, then to plastics micro- in oyster into
Row dyeing.
(1) dyeing of plastics preliminary experiment：A fritter plastics, are shredded on clip polybag, are put into equipped with 9.5mL methanol
In beaker, 0.5mL (3- aminopropyls) trimethoxy silane is added in liquid-transfering gun, after standing 12 hours, beaker liquid is discarded, and
It is cleaned 2 to 3 times with methanol, (3- aminopropyls) trimethoxy silane (SILVER REAGENT, this reagent Co., Ltd of Adama) is gone completely
It removes, then (FITC, the neat cloud biotechnology in Guangzhou are limited for the methanol of addition submergence frosting and micro fluorescein isothiocynate
Company), it dyes about 4 hours, finally with a large amount of ultrapure waters, preserves to be checked.Plastics are placed in clean glass slide with tweezers
On, use microexamination.As shown in Fig. 2, form of the plastics in 4 times of light fields of microscope is clearly observable, if seen under details in a play not acted out on stage, but told through dialogues
It examines, then not it is observed that plastics；As shown in figure 3, this is form of the plastics of the upper FITC of modification under 4 times of details in a play not acted out on stage, but told through dialogues of microscope, clearly
It is clear to observe that there is apparent green fluorescence at edge, illustrate that the silanization preliminary experiment of plastics is feasible.
(2) micro- dyeing of plastics preliminary experiment：The supernatant liquor 5mL of sample after being stood 12 hours in 1.2.4 is taken, uses 0.22 μm
Organic miillpore filter (upper Asia scavenging material factory of Haixing County) is filtered, and rinsed 2 to 3 times with methanol, and organic miillpore filter is added
Enter in the beaker equipped with methanol, methanol submerges organic miillpore filter, and seals up beaker mouth with preservative film, carries out 30 seconds super
Sound discards organic miillpore filter, and add in micro fluorescein isothiocynate (FITC) in beaker after ultrasonic, stands dyeing 4
Hour, it filters, with a large amount of ultrapure waters, is stored in be checked in ultra-pure water.20 μ L solution to be checked are drawn with liquid-transfering gun, draw 3
It is secondary, carry out parallel laboratory test；
Fig. 4 is the form that micro- dyeing of plastics is clearly observable according to this micro- dyeing of plastics preliminary experiment.But this scheme is unfavorable
It is counted in micro- plastics, above-mentioned image is only partial lens, and artificial counting error is larger, and therefore, sample concentration after digestion is carried
Height reduces the volume drawn during microexamination, so that drop is completely presented under single camera lens, and coordinates computer software meter
Number, thus 1.3.1 experimental programs (3) are designed, oyster is directly filtered after nitric acid digestion to 10mL ultra-pure waters, is not added full
Be layered with NaCL solution, dye before oscillation shake up, draw 5mL sample liquids dyed, micro- sem observation then draw 0.5 μ L with
Convenient for can be in the single camera lens complete observation of microscope.
(3) micro- dyeing of plastics experiment：It is another that an oyster (9.5g) is taken to be digested according to the method for 1.2.2 experiments 4, it takes out
Filter will filter the miillpore filter of completion, be preserved in ultrasound to the ultra-pure water of 10mL.Oscillation, shakes up, and draws 5mL liquid, uses
The 0.22 organic miillpore filters of μ L are filtered, and are rinsed 2 to 3 times with methanol, and organic miillpore filter is added in the burning equipped with methanol
In cup, methanol submerges organic miillpore filter, and seals up beaker mouth with preservative film, carries out the ultrasound of 30 seconds, having after ultrasound
Machine miillpore filter discards, and adds in micro fluorescein isothiocynate (FITC) in beaker, stands dyeing 4 hours, filters, with big
Ultrapure water is measured, is stored in be checked in ultra-pure water.
(a), (b), (c) are the experimental results of micro- dyeing of plastics in Fig. 5, and using computer software count, be from
After the digestion of 9.5g oysters filters, in 10mL ultra-pure water solutions, the sample liquid for drawing 5mL is dyed ultrasound, during microexamination
Draw 0.5 μ L.The quantity for scheming (a) is 27, and figure (b) quantity is 33, and the quantity of figure (c) is 35, and it is 32 to be averaged,
And it is calculated according to extension rate：
The oyster, which can be calculated, the micro- plastics of 67368/g
The composition detection of micro- plastics in 1.4 oysters
1.4.1 FTIR spectrum detects
(1) sample concentration：An oyster (13.44g) is taken to be digested according to the method for 1.2.2 experiments 4, filters, will filter
The miillpore filter of completion, ultrasound stand 12 hours in the ultra-pure water of 10mL, 1mL supernatant liquids are drawn in centrifuge tube, 80
About 0.2mL is dried at DEG C, is preserved to be checked.
(2) appropriate potassium bromide solid particle is taken to be ground to powdered, (day victory is won up to development in science and technology in Tianjin using compression mold
Co., Ltd) tablet to be checked is made, after 3 minutes dry in infrared flash dryer, the machine on FTIR spectrum that carries out is surveyed
It tries (TENSOR27 infrared spectrometers, German Brooker infrared gear Co., Ltd).
(3) appropriate potassium bromide solid particle is taken to be ground to powdered, the sample liquids concentrated in (1) is added in, with powder one
Play grinding after a certain period of time, it is 3 minutes dry in infrared flash dryer, tablet to be checked is made, then red using compression mold
It is 1 minute dry in outer flash dryer, carry out FTIR spectrum examination with computer.
Fig. 6 show the infrared spectrum figure of sample, only has apparent characteristic peak in 1400cm-1, according to Liu Rongzhong etc.
People's infrared spectrum identification plastics document is [with infrared spectrum identification plastic components [J] plastics science and technology, 2008,36 (6):72-77] it is aobvious
Showing (Fig. 7), polypropylene (PP) and polyvinyl chloride (PVC) have characteristic peak in 1400cm-1, and as polyethylene (PE), polystyrene
(PS), the common plastics such as polytetrafluoroethylene (PTFE) (PTFE) are then here without characteristic peak.Micro- plastics be in ocean, by light,
Under the action of the factors such as heat, biology, therefore the plastic flakes that decompose, physics and chemical characteristic may also change,
Functional group is different from common plastics, the Partial Feature peak of only former plastics.The sample only has under FTIR spectrum
There is obvious characteristic peak at 1400cm-1, i.e., it may be the micro- plastics formed by polypropylene (PP) or polyvinyl chloride (PVC) decomposition,
Or be both to have, therefore, for the oyster sample (13.44g), speculate that micro- plastic components therein are poly- herein
Propylene (PP) or polyvinyl chloride (PVC).
In addition, polypropylene (PP) and polyvinyl chloride (PVC) are all the common plastics of daily life.Polypropylene (PP) can be used in
The engineerings accessory such as automobile, electrical, weaving, daily necessities, medical device, construction material etc. also occur, and wherein fishing is restricted
Rope, strap, tie string, woven bag when polypropylene (PP) plastic products in ocean because operation due to lose or discarded, be all
Common behavior；Similarly, polyvinyl chloride (PVC) can also occur in construction material, daily necessities, packaging material, wherein moulding
The edible plastics such as material bag, mineral water bottle, beverage bottle container appeared in available to, discarded plastic bottle it is more common in ocean,
The moral consciousness for being not only people is inadequate, while plastic bottle is sent into ocean by the factor for also having force majeure.Above-mentioned example, all
It is to give in this paper oysters containing micro- plastics into component polypropylene (PP) or polyvinyl chloride (PVC), one more fully rationally
Property infer.
1.4.2 Electron microscopy
(1) is taken in 1.4.1 to preserve sample to be checked, inspection.For sample (13.44g), in electron microscope, (S4800 is swept Fig. 8
Retouch electron microscope, Hitachi Co., Ltd) under image, (a) (c) two figure has apparent plastic flakes feature, has
The plastic pieces of fracture, (b) figure have the plastic flakes in film-form, it may be possible to be decomposed, also further demonstrated that by polybag
Contain the inference of polychloroprene (PVC) in 4.3.1.
This research carries out wet digesting, a series of extraction experiments such as degreasing and its optimization, together using oyster as experimental subjects
When by micro- plastics of extraction using silyation technigue dye, to its quantity carry out preresearch estimates, also pass through FTIR spectrum
Technology and electron microscope find the type shape characteristic of micro- plastics.Result of study is as follows：
(1) extraction scheme optimized impregnates the oyster sample of the about 12g HNO3 of 25mL 12 hours, and heating 2 is small
When, and Surfactant SDS solution has been added to rinse filter with 80 DEG C while hot, rear 80 DEG C of ultrapure waters
Totally, if necessary to concentration, calculating, then in ultrasound to 10mL ultra-pure waters, such as it is detached, then can add in 10mL saturation chlorinations
Sodium solution.
(2) best Staining Protocol, the sample that cancellationization is completed is ultrasonic in the ultra-pure water of 10mL, and filters to organic
On miillpore filter, according to 19:1 ratio adds in methanol and (3- aminopropyls) trimethoxy silane modifies amino to micro- plastics, then
After secondary suction filtration, dyed with micro fluorescein isothiocynate (FITC), and in fluorescence microscopy Microscopic observation.This experiment oyster
As a result it is the micro- plastics of 67368/g.
(3) best characterization scheme, the sample that cancellationization is completed is ultrasonic in the ultra-pure water of 10mL, takes 1~2mL 80
It is dried at DEG C, sample concentration, and carries out FTIR spectrum detection and electron microscope characterization.The experimental results showed that this
Experiment detection oyster contain polypropylene (PP) or polyvinyl chloride (PVC), or both all have.
The foregoing is merely presently preferred embodiments of the present invention, is not intended to limit the invention.It is all the present invention spirit and
Any modification, replacement and improvement within principle should all be included in the protection scope of the present invention.
Claims (10)
1. a kind of method for detecting micro- plastics in Organism Samples, including：
S100, the step of micro- plastics are extracted from Organism Samples, including：
S110, resolution：Organism Samples with the nitric acid of a concentration of 50-70% are impregnated 6-18 hours at room temperature, are heated to 60-
It 95 DEG C and maintains 1-4 hours, the Organism Samples cleared up；
S120, degreasing：The Organism Samples of resolution obtained in step S110 are used to 60-95 DEG C of surfactant solution while hot
It rinses and filters, obtain micro- plastics of degreasing；
The water of 60-95 DEG C of micro- plastics of the degreasing obtained in step S120 is rinsed and filtered, the micro- modeling cleaned by S130
Material.
2. the method for micro- plastics, wherein step S100 are further included in the detection organism according to claim 1：By what is obtained
Micro- plastics of cleaning are added to the water, and are then ultrasonically treated, obtain micro- plastics suspension.
3. it is according to claim 1 it is described detection organism in micro- plastics method, wherein it is described suction filtration be with miillpore filter into
Capable suction filtration.
4. the method for micro- plastics, wherein step S100 are further included in the detection organism according to claim 1：
S140, classification：Micro- plastics are pressed into density classification with the different solution of density；Preferably, it is 1g/ that the solution, which is selected from density,
cm3Water, density 1.19g/cm3Saturated sodium-chloride water solution, density 0.91g/cm358.4% ethanol water, close
It spends for 0.925g/cm355.4% ethanol water and density be 1.27g/cm327.5% calcium chloride water.
5. the method for micro- plastics in the detection organism as claimed in one of claims 1-4, wherein the surface-active
Agent is lauryl sodium sulfate.
6. the method for micro- plastics in the detection organism as claimed in one of claims 1-4, wherein the organism is
Oyster.
7. the method for micro- plastics, further includes in the detection organism as claimed in one of claims 1-4：
Micro- plastics of the cleaning obtained in step S100 are carried out infrared spectrum detection or electron microscope characterize by S400.
8. the method for micro- plastics, further includes in the detection organism as claimed in one of claims 1-4：
S200 dyes micro- plastics of the cleaning obtained in step S100, including：With amino silicone and micro- plastic contacts
To obtain micro- plastics with amino, then dyed with fluorescein isothiocynate with micro- plastic contacts with amino
Micro- plastics.
9. the method for micro- plastics in the detection organism according to claim 8, wherein in step s 200, the amino silicone
Oxygen alkane is (3- aminopropyls) trimethoxy silane, according to 19:1 ratio adds in methanol and (3- aminopropyls) trimethoxy silane
To obtain micro- plastics with amino in micro- plastics.
10. the method for micro- plastics, further includes in the detection organism according to claim 8：
S300 with micro- plastics of the dyeing obtained in microscope observation of steps S200, and counts the particle of micro- plastics.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711178353.0A CN108181154B (en) | 2017-11-23 | 2017-11-23 | Method for detecting micro-plastics in organism |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711178353.0A CN108181154B (en) | 2017-11-23 | 2017-11-23 | Method for detecting micro-plastics in organism |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN108181154A true CN108181154A (en) | 2018-06-19 |
| CN108181154B CN108181154B (en) | 2023-04-18 |
Family
ID=62545235
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201711178353.0A Active CN108181154B (en) | 2017-11-23 | 2017-11-23 | Method for detecting micro-plastics in organism |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108181154B (en) |
Cited By (15)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109253952A (en) * | 2018-11-08 | 2019-01-22 | 深圳市美信检测技术股份有限公司 | The analysis method of particle in a kind of drinking water |
| CN109253907A (en) * | 2018-10-17 | 2019-01-22 | 大连理工大学 | A method of micro- plastics in quickly auxiliary detection environmental water sample are dyed using Nile red |
| CN109900886A (en) * | 2019-02-27 | 2019-06-18 | 南阳师范学院 | Whether the method for micro- plastics is contained in a kind of detection feces of livestock and poultry |
| CN110108629A (en) * | 2019-06-12 | 2019-08-09 | 深圳市美信分析技术有限公司 | The detection method of micro- plastic grain in a kind of chemical product |
| CN110243642A (en) * | 2019-06-19 | 2019-09-17 | 南京大学 | The method of micro- plastics is extracted in a kind of human and animal excreta |
| CN110646334A (en) * | 2019-09-29 | 2020-01-03 | 华东理工大学 | Rapid analysis method for small-size micro-plastic in water sample |
| CN111006927A (en) * | 2019-12-27 | 2020-04-14 | 山东师范大学 | Method for reducing interference of fat residue on fish body micro-plastic detection |
| CN111337327A (en) * | 2020-03-26 | 2020-06-26 | 河北大学 | Method and equipment for extracting farmland soil micro-plastic |
| CN111504741A (en) * | 2020-04-22 | 2020-08-07 | 山东师范大学 | Method for detecting micro-plastics in fish body |
| CN112525635A (en) * | 2020-11-20 | 2021-03-19 | 哈尔滨工业大学（深圳） | Method for extracting micro-plastic |
| CN113670779A (en) * | 2021-07-26 | 2021-11-19 | 杭州娃哈哈集团有限公司 | Method for detecting and evaluating distribution of micro-plastics in packaged drinking water based on infrared imaging technology |
| CN113984703A (en) * | 2021-11-01 | 2022-01-28 | 温州科技职业学院 | Device and method for detecting micro-plastic surface adsorbates in sea shrimps |
| CN114252427A (en) * | 2021-12-10 | 2022-03-29 | 暨南大学 | Method for effectively analyzing micro-plastics in grease-containing food sample |
| CN114486476A (en) * | 2021-12-27 | 2022-05-13 | 江苏省疾病预防控制中心(江苏省公共卫生研究院) | Pretreatment method for detecting polyethylene micro-plastic in liquid sample and application thereof |
| CN117007570A (en) * | 2023-10-08 | 2023-11-07 | 海南热带海洋学院 | Method for detecting microplastic in marine organism |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US11879835B1 (en) | 2023-08-11 | 2024-01-23 | King Faisal University | Device for detecting toxic elements in plastic products and determining potential health risks |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105651748A (en) * | 2016-01-18 | 2016-06-08 | 南京大学 | Method for quantitatively analyzing enrichment and distribution of micro plastics in aquatic organisms |
| CN106645049A (en) * | 2016-09-30 | 2017-05-10 | 大连海洋大学 | Method for detecting plastic content of marine organism |
| CN107160585A (en) * | 2017-07-06 | 2017-09-15 | 南京大学 | A kind of method for preparing graininess and the micro- plastics of laminar fluorescence labeling |
-
2017
- 2017-11-23 CN CN201711178353.0A patent/CN108181154B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105651748A (en) * | 2016-01-18 | 2016-06-08 | 南京大学 | Method for quantitatively analyzing enrichment and distribution of micro plastics in aquatic organisms |
| CN106645049A (en) * | 2016-09-30 | 2017-05-10 | 大连海洋大学 | Method for detecting plastic content of marine organism |
| CN107160585A (en) * | 2017-07-06 | 2017-09-15 | 南京大学 | A kind of method for preparing graininess and the micro- plastics of laminar fluorescence labeling |
Non-Patent Citations (2)
| Title |
|---|
| JEAN-PIERRE W. DESFORGES ET AL.: "Ingestion of Microplastics by Zooplankton in the Northeast Pacific Ocean", 《ARCH ENVIRON CONTAM TOXICOL》 * |
| ROBIN LENZ ET AL.: ""A critical assessment of visual identification of marine microplastic using Raman spectroscopy for analysis improvement", 《MARINE POLLUTION BULLETIN》 * |
Cited By (21)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109253907B (en) * | 2018-10-17 | 2020-11-13 | 大连理工大学 | Method for rapidly and auxiliarily detecting micro-plastic in water environment sample by using Nile red dyeing |
| CN109253907A (en) * | 2018-10-17 | 2019-01-22 | 大连理工大学 | A method of micro- plastics in quickly auxiliary detection environmental water sample are dyed using Nile red |
| CN109253952A (en) * | 2018-11-08 | 2019-01-22 | 深圳市美信检测技术股份有限公司 | The analysis method of particle in a kind of drinking water |
| CN109900886A (en) * | 2019-02-27 | 2019-06-18 | 南阳师范学院 | Whether the method for micro- plastics is contained in a kind of detection feces of livestock and poultry |
| CN110108629A (en) * | 2019-06-12 | 2019-08-09 | 深圳市美信分析技术有限公司 | The detection method of micro- plastic grain in a kind of chemical product |
| CN110243642A (en) * | 2019-06-19 | 2019-09-17 | 南京大学 | The method of micro- plastics is extracted in a kind of human and animal excreta |
| CN110243642B (en) * | 2019-06-19 | 2021-05-11 | 南京大学 | Method for extracting micro-plastics from human and animal excreta |
| WO2020253036A1 (en) * | 2019-06-19 | 2020-12-24 | 南京大学 | Method for extracting micro plastic from human and animal excreta |
| CN110646334A (en) * | 2019-09-29 | 2020-01-03 | 华东理工大学 | Rapid analysis method for small-size micro-plastic in water sample |
| CN111006927A (en) * | 2019-12-27 | 2020-04-14 | 山东师范大学 | Method for reducing interference of fat residue on fish body micro-plastic detection |
| CN111337327A (en) * | 2020-03-26 | 2020-06-26 | 河北大学 | Method and equipment for extracting farmland soil micro-plastic |
| CN111504741A (en) * | 2020-04-22 | 2020-08-07 | 山东师范大学 | Method for detecting micro-plastics in fish body |
| CN112525635A (en) * | 2020-11-20 | 2021-03-19 | 哈尔滨工业大学（深圳） | Method for extracting micro-plastic |
| CN112525635B (en) * | 2020-11-20 | 2022-01-04 | 哈尔滨工业大学（深圳） | Method for extracting micro-plastic |
| CN113670779A (en) * | 2021-07-26 | 2021-11-19 | 杭州娃哈哈集团有限公司 | Method for detecting and evaluating distribution of micro-plastics in packaged drinking water based on infrared imaging technology |
| CN113984703A (en) * | 2021-11-01 | 2022-01-28 | 温州科技职业学院 | Device and method for detecting micro-plastic surface adsorbates in sea shrimps |
| CN113984703B (en) * | 2021-11-01 | 2023-12-12 | 温州科技职业学院 | Device and method for detecting adsorbate on surface of micro-plastic in sea shrimp |
| CN114252427A (en) * | 2021-12-10 | 2022-03-29 | 暨南大学 | Method for effectively analyzing micro-plastics in grease-containing food sample |
| CN114252427B (en) * | 2021-12-10 | 2024-08-13 | 暨南大学 | Method for effectively analyzing microplastic in oil-containing food sample |
| CN114486476A (en) * | 2021-12-27 | 2022-05-13 | 江苏省疾病预防控制中心(江苏省公共卫生研究院) | Pretreatment method for detecting polyethylene micro-plastic in liquid sample and application thereof |
| CN117007570A (en) * | 2023-10-08 | 2023-11-07 | 海南热带海洋学院 | Method for detecting microplastic in marine organism |
Also Published As
| Publication number | Publication date |
|---|---|
| CN108181154B (en) | 2023-04-18 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108181154A (en) | The detection method of micro- plastics in a kind of organism | |
| CN110715835A (en) | Method for separating micro-plastics in environmental soil or sediment based on combination of flotation and centrifugation | |
| Prata et al. | Methods for sampling and detection of microplastics in water and sediment: A critical review | |
| Lusher et al. | Testing of methodology for measuring microplastics in blue mussels (Mytilus spp) and sediments, and recommendations for future monitoring of microplastics (R & D-project) | |
| KR101035427B1 (en) | Filter container for extracting cells | |
| Guo et al. | Recent advances in spectroscopic techniques for the analysis of microplastics in food | |
| CN109238948A (en) | A method of micro- plastic density distribution in detection water environment deposit | |
| Delgado-Gallardo et al. | From sampling to analysis: A critical review of techniques used in the detection of micro-and nanoplastics in aquatic environments | |
| CN106769327A (en) | A kind of Double-layered cup and the method for inspection for collecting Helminthic Eggs | |
| Torres et al. | Historical microplastic records in marine sediments: current progress and methodological evaluation | |
| CN109506984A (en) | Micro- plastics sampling column, micro- plastics acquisition device and digestion procedure in situ | |
| Poulsen et al. | Palynological preparation techniques, a new Macerationtank-method and other modifications | |
| Piarulli et al. | An effective strategy for the monitoring of microplastics in complex aquatic matrices: Exploiting the potential of near infrared hyperspectral imaging (NIR-HSI) | |
| CN114608927A (en) | Method for extracting micro-plastics from soil by microwave digestion method | |
| CN111504741B (en) | Method for detecting micro-plastics in fish body | |
| CN107596735A (en) | A kind of preparation facilities and method of super-amphiphobic automatically cleaning water-oil separating material | |
| KeChi-Okafor et al. | Prevalence and characterisation of microfibres along the Kenyan and Tanzanian coast | |
| Mendes et al. | Microplastic in mangroves: A worldwide review of contamination in biotic and abiotic matrices | |
| CN105452849B (en) | The turbidity of fluid sample, the system and method for color and chlorinity are measured simultaneously | |
| CN103105342A (en) | Quantitative analysis method for yarns | |
| CN109187367B (en) | Device and method for detecting activity of microalgae in ballast water by using polarized light flow control sheet | |
| CN201583498U (en) | Civilian food safety test box | |
| CN204896298U (en) | It holds device to detect card based on food detects | |
| CN105076109A (en) | Mucus treating fluid and application thereof | |
| CN114577770A (en) | Full-size extraction and detection method for micro-plastic in deep-sea cold spring area sediment |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |