Quantification of fluorescent dyes in organ tissue samples via HPLC analysis
•Microspheres for blood perfusion measurements in pigs were extracted from brain, heart and kidney tissue samples.•Simultaneous quantification of four different microsphere types by extraction and separation of the encapsulated dyes.•Short run times of
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| Veröffentlicht in: | Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Analytical technologies in the biomedical and life sciences, 2018-01, Vol.1072, p.34-39 |
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| container_title | Journal of chromatography. B, Analytical technologies in the biomedical and life sciences |
| container_volume | 1072 |
| creator | Morsbach, S. García-Bardon, A. Kamuf, J. Müller, B. Beghersa, N. Mohr, K. Landfester, K. |
| description | •Microspheres for blood perfusion measurements in pigs were extracted from brain, heart and kidney tissue samples.•Simultaneous quantification of four different microsphere types by extraction and separation of the encapsulated dyes.•Short run times of |
| doi_str_mv | 10.1016/j.jchromb.2017.11.012 |
| format | Article |
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The determination of regional blood flow via the accumulation of fluorescent microspheres is a concept regularly used in medical research. Typically, the microbeads get extracted from the tissue of interest and are then quantified by measuring the absorption or fluorescence of the incorporated dyes without further separation from the medium. However, in that case the absorption spectra of different dyes can overlap when used simultaneously, leading to an overestimation of the concentration. Additionally, background absorption from the medium can be problematic. Therefore, a high performance liquid chromatography method for the simultaneous detection of four dyes (orange, crimson, yellow-green and red) incorporated in different microbeads in samples from biological media such as organ tissue (brain, heart and kidneys) was developed. Since for biological samples often a large sample size is required for sufficient statistics, the method was optimized to yield very short run times. With this method it was possible to detect very low concentrations of only one microsphere per gram of organ tissue. By applying this sensitive quantification technique, it was demonstrated that the application of microbeads for perfusion measurements might not be reliable due to different organ distributions in each animal.</description><identifier>ISSN: 1570-0232</identifier><identifier>EISSN: 1873-376X</identifier><identifier>DOI: 10.1016/j.jchromb.2017.11.012</identifier><identifier>PMID: 29132023</identifier><language>eng</language><publisher>Netherlands: Elsevier B.V</publisher><subject>Biological media ; Blood perfusion measurement ; Dye quantification ; Microbeads ; Reversed phase HPLC ; Tissue extraction</subject><ispartof>Journal of chromatography. B, Analytical technologies in the biomedical and life sciences, 2018-01, Vol.1072, p.34-39</ispartof><rights>2017 Elsevier B.V.</rights><rights>Copyright © 2017 Elsevier B.V. All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c365t-8b036c2c40d9c4635354e8144029c2be4e333720ef605e38dce401b121ec5a9a3</citedby><cites>FETCH-LOGICAL-c365t-8b036c2c40d9c4635354e8144029c2be4e333720ef605e38dce401b121ec5a9a3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.jchromb.2017.11.012$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3536,27903,27904,45974</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/29132023$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Morsbach, S.</creatorcontrib><creatorcontrib>García-Bardon, A.</creatorcontrib><creatorcontrib>Kamuf, J.</creatorcontrib><creatorcontrib>Müller, B.</creatorcontrib><creatorcontrib>Beghersa, N.</creatorcontrib><creatorcontrib>Mohr, K.</creatorcontrib><creatorcontrib>Landfester, K.</creatorcontrib><title>Quantification of fluorescent dyes in organ tissue samples via HPLC analysis</title><title>Journal of chromatography. B, Analytical technologies in the biomedical and life sciences</title><addtitle>J Chromatogr B Analyt Technol Biomed Life Sci</addtitle><description>•Microspheres for blood perfusion measurements in pigs were extracted from brain, heart and kidney tissue samples.•Simultaneous quantification of four different microsphere types by extraction and separation of the encapsulated dyes.•Short run times of <3min and a sensitivity of ∼1 sphere/g of tissue were achieved.•Microsphere concentrations in animals varied significantly, rendering microspheres problematic for perfusion measurements.
The determination of regional blood flow via the accumulation of fluorescent microspheres is a concept regularly used in medical research. Typically, the microbeads get extracted from the tissue of interest and are then quantified by measuring the absorption or fluorescence of the incorporated dyes without further separation from the medium. However, in that case the absorption spectra of different dyes can overlap when used simultaneously, leading to an overestimation of the concentration. Additionally, background absorption from the medium can be problematic. Therefore, a high performance liquid chromatography method for the simultaneous detection of four dyes (orange, crimson, yellow-green and red) incorporated in different microbeads in samples from biological media such as organ tissue (brain, heart and kidneys) was developed. Since for biological samples often a large sample size is required for sufficient statistics, the method was optimized to yield very short run times. With this method it was possible to detect very low concentrations of only one microsphere per gram of organ tissue. By applying this sensitive quantification technique, it was demonstrated that the application of microbeads for perfusion measurements might not be reliable due to different organ distributions in each animal.</description><subject>Biological media</subject><subject>Blood perfusion measurement</subject><subject>Dye quantification</subject><subject>Microbeads</subject><subject>Reversed phase HPLC</subject><subject>Tissue extraction</subject><issn>1570-0232</issn><issn>1873-376X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2018</creationdate><recordtype>article</recordtype><recordid>eNqFkF1LwzAUhoMobk5_gtJLb1pzkjRdr0SGOmGggoJ3IU1PNaUfM2kH-_dmbHrrVcI5z8l78hByCTQBCvKmTmrz5fq2SBiFLAFIKLAjMoV5xmOeyY_jcE8zGlPG2YSceV_TANKMn5IJy4Gz0JiS1euou8FW1ujB9l3UV1HVjL1Db7AbonKLPrKh7D51Fw3W-xEjr9t1E-obq6Ply2oR6U43W2_9OTmpdOPx4nDOyPvD_dtiGa-eH58Wd6vYcJkO8bygXBpmBC1zIyRPeSpwDkJQlhtWoEDOecYoVpKmyOelQUGhAAZoUp1rPiPX-3fXrv8e0Q-qtWHfptEd9qNXkEvBZC6ZCGi6R43rvXdYqbWzrXZbBVTtRKpaHUSqnUgFoILIMHd1iBiLFsu_qV9zAbjdAxg-urHolDcWO4OldWgGVfb2n4gfGVuGPw</recordid><startdate>20180101</startdate><enddate>20180101</enddate><creator>Morsbach, S.</creator><creator>García-Bardon, A.</creator><creator>Kamuf, J.</creator><creator>Müller, B.</creator><creator>Beghersa, N.</creator><creator>Mohr, K.</creator><creator>Landfester, K.</creator><general>Elsevier B.V</general><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>20180101</creationdate><title>Quantification of fluorescent dyes in organ tissue samples via HPLC analysis</title><author>Morsbach, S. ; García-Bardon, A. ; Kamuf, J. ; Müller, B. ; Beghersa, N. ; Mohr, K. ; Landfester, K.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c365t-8b036c2c40d9c4635354e8144029c2be4e333720ef605e38dce401b121ec5a9a3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2018</creationdate><topic>Biological media</topic><topic>Blood perfusion measurement</topic><topic>Dye quantification</topic><topic>Microbeads</topic><topic>Reversed phase HPLC</topic><topic>Tissue extraction</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Morsbach, S.</creatorcontrib><creatorcontrib>García-Bardon, A.</creatorcontrib><creatorcontrib>Kamuf, J.</creatorcontrib><creatorcontrib>Müller, B.</creatorcontrib><creatorcontrib>Beghersa, N.</creatorcontrib><creatorcontrib>Mohr, K.</creatorcontrib><creatorcontrib>Landfester, K.</creatorcontrib><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of chromatography. 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The determination of regional blood flow via the accumulation of fluorescent microspheres is a concept regularly used in medical research. Typically, the microbeads get extracted from the tissue of interest and are then quantified by measuring the absorption or fluorescence of the incorporated dyes without further separation from the medium. However, in that case the absorption spectra of different dyes can overlap when used simultaneously, leading to an overestimation of the concentration. Additionally, background absorption from the medium can be problematic. Therefore, a high performance liquid chromatography method for the simultaneous detection of four dyes (orange, crimson, yellow-green and red) incorporated in different microbeads in samples from biological media such as organ tissue (brain, heart and kidneys) was developed. Since for biological samples often a large sample size is required for sufficient statistics, the method was optimized to yield very short run times. With this method it was possible to detect very low concentrations of only one microsphere per gram of organ tissue. By applying this sensitive quantification technique, it was demonstrated that the application of microbeads for perfusion measurements might not be reliable due to different organ distributions in each animal.</abstract><cop>Netherlands</cop><pub>Elsevier B.V</pub><pmid>29132023</pmid><doi>10.1016/j.jchromb.2017.11.012</doi><tpages>6</tpages></addata></record> |
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| identifier | ISSN: 1570-0232 |
| ispartof | Journal of chromatography. B, Analytical technologies in the biomedical and life sciences, 2018-01, Vol.1072, p.34-39 |
| issn | 1570-0232 1873-376X |
| language | eng |
| recordid | cdi_proquest_miscellaneous_1964269624 |
| source | Elsevier ScienceDirect Journals |
| subjects | Biological media Blood perfusion measurement Dye quantification Microbeads Reversed phase HPLC Tissue extraction |
| title | Quantification of fluorescent dyes in organ tissue samples via HPLC analysis |
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