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生物体微塑料提取方法比选研究
Comparison of microplastic extraction methods from organisms
微塑料（<5mm）在淡水、海洋以及陆生生物体内被广泛检出，但由于提取方法不一致，限制了现有数据之间的可比性.本研究比选了4种消解方法对鱼胃等7种组织的消解效率，并通过显微镜检查、重量、傅里叶变换红外光谱（FT-IR）和拉曼光谱综合评估不同提取方法对10种环境微塑料形态、重量和光谱特征的影响.结果显示，消解效率依次为10% KOH > RIPA组织裂解液+蛋白酶K > 蛋白酶K > 30% H2O2.另外，4种消解处理后PA（聚酰胺）颗粒重量均显著增加，H2O2处理后PU（聚氨酯）颗粒颜色略有变化，可生物降解塑料溶于10% KOH，而其他塑料处理前后重量和形态均未变化；4种消解处理前后所有塑料的红外光谱与拉曼光谱均未发生改变.综合以上，推荐以10% KOH为消解液，50℃、180r/min温育振荡6h作为分离提取生物组织微塑料的首选方案.
Microplastics (<5mm) have been detected in freshwater, marine and terrestrial organisms widely, however, no harmonized method is available which limited the scientific value for these existing investigation data. The present study aimed to compare four kinds of digestion solutions for microplastic isolation from fish and clam tissue, and then to evaluate the effects on 10 kinds of typical plastics of four kinds of digestion solutions by using a group of indicators, including morphological observation, quality change, fourier transform infrared spectroscopy and Raman spectroscopy characteristics. The results showed that the digestive efficiencies (%) were followed as the order:10%KOH > RIPA tissue lysate + Proteinase K > Proteinase K > 30%H2O2. In addition, PA particle quality increased after treatments, and PU particle colour slight changed after H2O2 treatment. Interestingly, the biodegradable plastic particles were soluble in 10% KOH completely. As expected, the infrared spectrum and Raman spectrum of plastic particles did not change before and after treatments, and the plastic particles could be identified based on their spectrum, except for the biodegradable plastic. As a conclusion, the treatment method of 10% KOH at 50℃ and 180r/min incubated for 6hours will be recommended as the preferred method for microplastic isolation from biological tissues.
10% KOH / 分离 / 生物 / 微塑料 / 消解效率
10%KOH / digestion efficiency / isolation / microplastic / organism
[1] UNEP. Marine plastic debris and microplastics:global lessons and research to inspire action and guide policy change[Z]. 2018.
[2] Arthur C, Baker J, Bamford H (eds). Proceedings of the international research workshop on the occurrence, effects and fate of microplastic marine debris[R]. NOAA Technical Memorandum NOS-OR&R-30, 2008.
[3] Alimi O S, Farner B J, Hernandez L M, et al. Microplastics and nanoplastics in aquatic environments:aggregation, deposition, and enhanced contaminant transport[J]. Environmental Science and Technology, 2018,52(4):1704-1724.
[4] Dehaut A, Cassone A, Frère L, et al. Microplastics in seafood:Benchmark protocol for their extraction and characterization[J]. Environmental Pollution, 2016,215:223-233.
[5] Miller M E, Kroon F J, Motti C A. Recovering microplastics from marine samples:A review of current practices[J]. Marine Pollution Bulletin, 2017,123(1/2):6-18.
[6] Andrady A L. The plastic in microplastics:A review[J]. Marine Pollution Bulletin, 2017,119(1):12-22.
[7] Ogonowski M, Schür C, Jarsén Å, et al. The effects of natural and anthropogenic microparticles on individual fitness in Daphnia magna[J]. PLOS ONE, 2016,11(5):e155063.
[8] Rochman C M, Hoh E, Kurobe T, et al. Ingested plastic transfers hazardous chemicals to fish and induces hepatic stress[J]. Scientific Reports, 2013,3(1):1-7.
[9] 杨婧婧,徐笠,陆安祥,等.环境中微(纳米)塑料的来源及毒理学研究进展[J]. 环境化学, 2018,37(3):383-396. Yang J J, Xu L, Lu A X, et al. Research progress on the sources and toxicology of micro(nano) plastics in environment[J]. Environmental Chemistry, 2018,37(3):383-396.
[10] Cannon S M E, Lavers J L, Figueiredo B. Plastic ingestion by fish in the Southern Hemisphere:A baseline study and review of methods[J]. Marine Pollution Bulletin, 2016,107(1):286-291.
[11] Catarino A I, Thompson R, Sanderson W, et al. Development and optimization of a standard method for extraction of microplastics in mussels by enzyme digestion of soft tissues[J]. Environmental Toxicology and Chemistry, 2017,36(4):947-951.
[12] Cole M, Webb H, Lindeque P K, et al. Isolation of microplastics in biota-rich seawater samples and marine organisms[J]. Scientific Reports, 2015,4(1):1-8.
[13] Güven O, Gökda? K, Jovanovi? B, et al. Microplastic litter composition of the Turkish territorial waters of the Mediterranean Sea, and its occurrence in the gastrointestinal tract of fish[J]. Environmental Pollution, 2017,223:286-294.
[14] Cheung L, Lui C, Fok L. Microplastic contamination of wild and captive flathead grey mullet (Mugil cephalus)[J]. International Journal of Environmental Research and Public Health, 2018,15(4):597-607.
[15] Jabeen K, Su L, Li J, et al. Microplastics and mesoplastics in fish from coastal and fresh waters of China[J]. Environmental Pollution, 2017,221:141-149.
[16] Collard F, Gilbert B, Eppe G, et al. Detection of anthropogenic particles in fish stomachs:An isolation method adapted to identification by Raman spectroscopy[J]. Archives of Environmental Contamination and Toxicology, 2015,69(3):331-339.
[17] Ices. OSPAR request on development of a common monitoring protocol for plastic particles in fish stomachs and selected shellfish on the basis of existing fish disease surveys[R]. 2015.
[18] Avio C G, Gorbi S, Regoli F. Plastics and microplastics in the oceans:From emerging pollutants to emerged threat[J]. Marine Environmental Research, 2017,128:2-11.
[19] Van Cauwenberghe L, Janssen C R. Microplastics in bivalves cultured for human consumption[J]. Environmental Pollution, 2014,193:65-70.
[20] Takeichi T, Kitamura O. Detection of diatom in formalin-fixed tissue by proteinase K digestion[J]. Forensic Science International, 2009,190(1-3):19-23.
[21] Karami A, Golieskardi A, Choo C K, et al. A high-performance protocol for extraction of microplastics in fish[J]. Science of The Total Environment, 2017,578:485-494.
[22] 孙浩然.鱼体内微塑料检测方法研究[D]. 济南:山东师范大学, 2018. Sun H R. Research on micro-plastic detection methods in fish[D]. Jinan:Shandong Normal University, 2018.
[23] Karlsson T M, Vethaak A D, Almroth B C, et al. Screening for microplastics in sediment, water, marine invertebrates and fish:Method development and microplastic accumulation[J]. Marine Pollution Bulletin, 2017,122(1/2):403-408.
[24] 李晖,魏莉萍,王湘明,等.吸水率对尼龙66力学性能的影响研究[J]. 工程塑料应用, 2008,(8):64-67. Li H, Wei L P, Wang X M, et al. Effect of water absorption rate on mechanical properties of nylon 66[J]. Engineering Plastics application, 2008,(8):64-67.
[25] Enders K, Lenz R, Beer S, et al. Extraction of microplastic from biota:Recommended acidic digestion destroys common plastic polymers[J]. ICES Journal of Marine Science:Journal du Conseil, 2016:fsw173:1-6.
[26] Roch S, Brinker A. Rapid and efficient method for the detection of microplastic in the gastrointestinal tract of fishes[J]. Environmental Science and Technology, 2017,51(8):4522.
[27] Simon M, van Alst N, Vollertsen J. Quantification of microplastic mass and removal rates at wastewater treatment plants applying Focal Plane Array (FPA)-based Fourier Transform Infrared (FT-IR) imaging[J]. Water Research, 2018,142:1-9.
[28] Li J, Qu X, Su L, et al. Microplastics in mussels along the coastal waters of China[J]. Environmental Pollution, 2016,214:177-184.
[29] 翁诗甫.傅里叶变换红外光谱分析[M]. 化学工业出版社, 2005:389. Weng S Z. Fourier transform infrared spectroscopy[M]. Chemical Industry Press, 2005:389.
[30] Claessens M, Van Cauwenberghe L, Vandegehuchte M B, et al. New techniques for the detection of microplastics in sediments and field collected organisms[J]. Marine Pollution Bulletin, 2013,70(1/2):227-233.
[31] Lenz R, Enders K, Stedmon C A, et al. A critical assessment of visual identification of marine microplastic using Raman spectroscopy for analysis improvement[J]. Marine Pollution Bulletin, 2015,100(1):82-91.
[32] Araujo C F, Nolasco M M, Ribeiro A M P, et al. Identification of microplastics using Raman spectroscopy:Latest developments and future prospects[J]. Water Research, 2018,142:426-440.
[33] Borman S A. Nonlinear Raman spectroscopy[J]. Analytical Chemistry, 1982,54(9):1021-1026.
国家自然科学基金资助项目（21577137）；国家重点研发计划（2016YFC1402206）
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