Elizabeth C. Minor, Uttam D. Gomes, Kathryn M. Schreiner, Nicole J. Poulton, Erik Hendrickson, Melissa A. Maurer-Jones
{"title":"苏必利尔湖中的微小塑料颗粒:尼罗河红染色、流式细胞仪和高温分解气相色谱-质谱联用初步研究","authors":"Elizabeth C. Minor, Uttam D. Gomes, Kathryn M. Schreiner, Nicole J. Poulton, Erik Hendrickson, Melissa A. Maurer-Jones","doi":"10.1002/lom3.10582","DOIUrl":null,"url":null,"abstract":"<p>Microplastic particles (< 5 mm) are now found throughout earth's ecosystems, with smaller microplastics often showing greater impacts on organismal health than larger ones. Unfortunately, there are no readily available analytical approaches that can couple microplastics enumeration and polymer determination for smaller microplastics (< 10 <i>μ</i>m), and 1–20 <i>μ</i>m particles are difficult to quantify with existing techniques. This study presents a method using Nile red (NR) staining and flow cytometry (FCM) to quantify and isolate small microplastic particles for subsequent identification by pyrolysis gas chromatography–mass spectrometry (pyGCMS). Results using standard plastic particles showed that FCM sorting can provide sufficient material for pyGCMS analyses; the polymer composition remains identifiable after the processing steps. The post-sorting concentration step yielded recovery of 58%–83% of the original plastic polymer mass. Analysis of a mixed plastic standard solution showed no significant difference in plastic counts obtained by microscopy and FCM, although blank correction reduces the FCM counts to 62% of the microscopy counts. The applicability of NR staining and FCM was demonstrated through analysis of small microplastic particles (5–45 <i>μ</i>m) from Lake Superior surface water samples, which showed particle abundances two to three orders of magnitude higher than particles > 100 <i>μ</i>m that were counted using FTIR microscopy. PyGCMS analysis of a test lake sample showed the presence of polyethylene in this small size fraction. Careful attention to blanks and longer FCM sorting times (> 2 h) are recommended for successful analysis of natural aquatic samples processed by this approach.</p>","PeriodicalId":18145,"journal":{"name":"Limnology and Oceanography: Methods","volume":"21 12","pages":"800-813"},"PeriodicalIF":2.1000,"publicationDate":"2023-11-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://aslopubs.onlinelibrary.wiley.com/doi/epdf/10.1002/lom3.10582","citationCount":"0","resultStr":"{\"title\":\"Small microplastic particles in Lake Superior: A preliminary study coupling Nile red staining, flow cytometry and pyrolysis gas chromatography–mass spectrometry\",\"authors\":\"Elizabeth C. Minor, Uttam D. Gomes, Kathryn M. Schreiner, Nicole J. Poulton, Erik Hendrickson, Melissa A. Maurer-Jones\",\"doi\":\"10.1002/lom3.10582\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p>Microplastic particles (< 5 mm) are now found throughout earth's ecosystems, with smaller microplastics often showing greater impacts on organismal health than larger ones. Unfortunately, there are no readily available analytical approaches that can couple microplastics enumeration and polymer determination for smaller microplastics (< 10 <i>μ</i>m), and 1–20 <i>μ</i>m particles are difficult to quantify with existing techniques. This study presents a method using Nile red (NR) staining and flow cytometry (FCM) to quantify and isolate small microplastic particles for subsequent identification by pyrolysis gas chromatography–mass spectrometry (pyGCMS). Results using standard plastic particles showed that FCM sorting can provide sufficient material for pyGCMS analyses; the polymer composition remains identifiable after the processing steps. The post-sorting concentration step yielded recovery of 58%–83% of the original plastic polymer mass. Analysis of a mixed plastic standard solution showed no significant difference in plastic counts obtained by microscopy and FCM, although blank correction reduces the FCM counts to 62% of the microscopy counts. The applicability of NR staining and FCM was demonstrated through analysis of small microplastic particles (5–45 <i>μ</i>m) from Lake Superior surface water samples, which showed particle abundances two to three orders of magnitude higher than particles > 100 <i>μ</i>m that were counted using FTIR microscopy. PyGCMS analysis of a test lake sample showed the presence of polyethylene in this small size fraction. 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Small microplastic particles in Lake Superior: A preliminary study coupling Nile red staining, flow cytometry and pyrolysis gas chromatography–mass spectrometry
Microplastic particles (< 5 mm) are now found throughout earth's ecosystems, with smaller microplastics often showing greater impacts on organismal health than larger ones. Unfortunately, there are no readily available analytical approaches that can couple microplastics enumeration and polymer determination for smaller microplastics (< 10 μm), and 1–20 μm particles are difficult to quantify with existing techniques. This study presents a method using Nile red (NR) staining and flow cytometry (FCM) to quantify and isolate small microplastic particles for subsequent identification by pyrolysis gas chromatography–mass spectrometry (pyGCMS). Results using standard plastic particles showed that FCM sorting can provide sufficient material for pyGCMS analyses; the polymer composition remains identifiable after the processing steps. The post-sorting concentration step yielded recovery of 58%–83% of the original plastic polymer mass. Analysis of a mixed plastic standard solution showed no significant difference in plastic counts obtained by microscopy and FCM, although blank correction reduces the FCM counts to 62% of the microscopy counts. The applicability of NR staining and FCM was demonstrated through analysis of small microplastic particles (5–45 μm) from Lake Superior surface water samples, which showed particle abundances two to three orders of magnitude higher than particles > 100 μm that were counted using FTIR microscopy. PyGCMS analysis of a test lake sample showed the presence of polyethylene in this small size fraction. Careful attention to blanks and longer FCM sorting times (> 2 h) are recommended for successful analysis of natural aquatic samples processed by this approach.
期刊介绍:
Limnology and Oceanography: Methods (ISSN 1541-5856) is a companion to ASLO''s top-rated journal Limnology and Oceanography, and articles are held to the same high standards. In order to provide the most rapid publication consistent with high standards, Limnology and Oceanography: Methods appears in electronic format only, and the entire submission and review system is online. Articles are posted as soon as they are accepted and formatted for publication.
Limnology and Oceanography: Methods will consider manuscripts whose primary focus is methodological, and that deal with problems in the aquatic sciences. Manuscripts may present new measurement equipment, techniques for analyzing observations or samples, methods for understanding and interpreting information, analyses of metadata to examine the effectiveness of approaches, invited and contributed reviews and syntheses, and techniques for communicating and teaching in the aquatic sciences.