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Determination of Total Glutathione in Dried Blood Spot Samples Using a High-Performance Liquid Chromatography
A method is described for the determination of total glutathione (TGSH) in dried blood spot (DBS) samples using high-performance liquid chromatography with fluorescence detection. Whole blood and DBS samples were obtained from a group of blood donors. After GSH reduction with dithiothreitol and prot...
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Published in: | Journal of chromatographic science 2015-07, Vol.53 (6), p.879-885 |
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container_title | Journal of chromatographic science |
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creator | Kanďár, Roman Štramová, Xenie Drábková, Petra Brandtnerová, Martina |
description | A method is described for the determination of total glutathione (TGSH) in dried blood spot (DBS) samples using high-performance liquid chromatography with fluorescence detection. Whole blood and DBS samples were obtained from a group of blood donors. After GSH reduction with dithiothreitol and protein precipitation with ethanol, the samples were derivatized with naphthalene-2,3-dicarboxaldehyde to form a very stable, highly fluorescent derivative. For the separation, a reversed phase HPLC method was used. The mixture of ethanol and deionized water (8 : 92, v/v) was used as a mobile phase. The analytical performance of this method was satisfactory: the intra- and interassay coefficients of variation were below 10%. Quantitative recoveries from spiked DBS samples were between 98.3 and 103.6%. The presented method is inexpensive and suitable for clinical testing purposes. |
doi_str_mv | 10.1093/chromsci/bmu135 |
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Whole blood and DBS samples were obtained from a group of blood donors. After GSH reduction with dithiothreitol and protein precipitation with ethanol, the samples were derivatized with naphthalene-2,3-dicarboxaldehyde to form a very stable, highly fluorescent derivative. For the separation, a reversed phase HPLC method was used. The mixture of ethanol and deionized water (8 : 92, v/v) was used as a mobile phase. The analytical performance of this method was satisfactory: the intra- and interassay coefficients of variation were below 10%. Quantitative recoveries from spiked DBS samples were between 98.3 and 103.6%. The presented method is inexpensive and suitable for clinical testing purposes.</description><identifier>ISSN: 0021-9665</identifier><identifier>EISSN: 1945-239X</identifier><identifier>DOI: 10.1093/chromsci/bmu135</identifier><identifier>PMID: 25344838</identifier><language>eng</language><publisher>United States: Oxford University Press</publisher><subject>Adolescent ; Adult ; Chromatography, High Pressure Liquid - methods ; Dried Blood Spot Testing - methods ; Female ; Glutathione - blood ; Glutathione - chemistry ; Humans ; Limit of Detection ; Linear Models ; Male ; Methanol ; Middle Aged ; Reproducibility of Results ; Young Adult</subject><ispartof>Journal of chromatographic science, 2015-07, Vol.53 (6), p.879-885</ispartof><rights>The Author 2014. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com 2014</rights><rights>The Author 2014. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c373t-4a5a52758078429c06881cacf19c2d8314e61326a2897f05531b45c02813c373</citedby><cites>FETCH-LOGICAL-c373t-4a5a52758078429c06881cacf19c2d8314e61326a2897f05531b45c02813c373</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/25344838$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Kanďár, Roman</creatorcontrib><creatorcontrib>Štramová, Xenie</creatorcontrib><creatorcontrib>Drábková, Petra</creatorcontrib><creatorcontrib>Brandtnerová, Martina</creatorcontrib><title>Determination of Total Glutathione in Dried Blood Spot Samples Using a High-Performance Liquid Chromatography</title><title>Journal of chromatographic science</title><addtitle>J Chromatogr Sci</addtitle><description>A method is described for the determination of total glutathione (TGSH) in dried blood spot (DBS) samples using high-performance liquid chromatography with fluorescence detection. Whole blood and DBS samples were obtained from a group of blood donors. After GSH reduction with dithiothreitol and protein precipitation with ethanol, the samples were derivatized with naphthalene-2,3-dicarboxaldehyde to form a very stable, highly fluorescent derivative. For the separation, a reversed phase HPLC method was used. The mixture of ethanol and deionized water (8 : 92, v/v) was used as a mobile phase. The analytical performance of this method was satisfactory: the intra- and interassay coefficients of variation were below 10%. Quantitative recoveries from spiked DBS samples were between 98.3 and 103.6%. The presented method is inexpensive and suitable for clinical testing purposes.</description><subject>Adolescent</subject><subject>Adult</subject><subject>Chromatography, High Pressure Liquid - methods</subject><subject>Dried Blood Spot Testing - methods</subject><subject>Female</subject><subject>Glutathione - blood</subject><subject>Glutathione - chemistry</subject><subject>Humans</subject><subject>Limit of Detection</subject><subject>Linear Models</subject><subject>Male</subject><subject>Methanol</subject><subject>Middle Aged</subject><subject>Reproducibility of Results</subject><subject>Young Adult</subject><issn>0021-9665</issn><issn>1945-239X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2015</creationdate><recordtype>article</recordtype><recordid>eNqFkM9LwzAYQIMobk7P3iRHEeryo2nTo266CQOFTfBWsjTdIk3TJelh_70tdV49hS-87_HxALjF6BGjjE7l3lnjpZ5uTYspOwNjnMUsIjT7OgdjhAiOsiRhI3Dl_Xc_Ys4uwYgwGsec8jEwcxWUM7oWQdsa2hJubBAVXFRtEGHf_Smoazh3WhXwubK2gOvGBrgWpqmUh59e1zso4FLv9tGHcqV1RtRSwZU-tLqAs_5AEezOiWZ_vAYXpai8uvl9J2Dz-rKZLaPV--Jt9rSKJE1piGLBBCMp4yjlMckkSjjHUsgSZ5IUnOJYJZiSRBCepSVijOJtzCQiHNPeMAH3g7Zx9tAqH3KjvVRVJWplW5_jhKc0zhDlHTodUOms906VeeO0Ee6YY5T3ifNT4nxI3G3c_crbrVHFH39q2gEPA2Db5l_bD7KjiKA</recordid><startdate>201507</startdate><enddate>201507</enddate><creator>Kanďár, Roman</creator><creator>Štramová, Xenie</creator><creator>Drábková, Petra</creator><creator>Brandtnerová, Martina</creator><general>Oxford University Press</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>201507</creationdate><title>Determination of Total Glutathione in Dried Blood Spot Samples Using a High-Performance Liquid Chromatography</title><author>Kanďár, Roman ; Štramová, Xenie ; Drábková, Petra ; Brandtnerová, Martina</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c373t-4a5a52758078429c06881cacf19c2d8314e61326a2897f05531b45c02813c373</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2015</creationdate><topic>Adolescent</topic><topic>Adult</topic><topic>Chromatography, High Pressure Liquid - methods</topic><topic>Dried Blood Spot Testing - methods</topic><topic>Female</topic><topic>Glutathione - blood</topic><topic>Glutathione - chemistry</topic><topic>Humans</topic><topic>Limit of Detection</topic><topic>Linear Models</topic><topic>Male</topic><topic>Methanol</topic><topic>Middle Aged</topic><topic>Reproducibility of Results</topic><topic>Young Adult</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Kanďár, Roman</creatorcontrib><creatorcontrib>Štramová, Xenie</creatorcontrib><creatorcontrib>Drábková, Petra</creatorcontrib><creatorcontrib>Brandtnerová, Martina</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of chromatographic science</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Kanďár, Roman</au><au>Štramová, Xenie</au><au>Drábková, Petra</au><au>Brandtnerová, Martina</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Determination of Total Glutathione in Dried Blood Spot Samples Using a High-Performance Liquid Chromatography</atitle><jtitle>Journal of chromatographic science</jtitle><addtitle>J Chromatogr Sci</addtitle><date>2015-07</date><risdate>2015</risdate><volume>53</volume><issue>6</issue><spage>879</spage><epage>885</epage><pages>879-885</pages><issn>0021-9665</issn><eissn>1945-239X</eissn><abstract>A method is described for the determination of total glutathione (TGSH) in dried blood spot (DBS) samples using high-performance liquid chromatography with fluorescence detection. Whole blood and DBS samples were obtained from a group of blood donors. After GSH reduction with dithiothreitol and protein precipitation with ethanol, the samples were derivatized with naphthalene-2,3-dicarboxaldehyde to form a very stable, highly fluorescent derivative. For the separation, a reversed phase HPLC method was used. The mixture of ethanol and deionized water (8 : 92, v/v) was used as a mobile phase. The analytical performance of this method was satisfactory: the intra- and interassay coefficients of variation were below 10%. Quantitative recoveries from spiked DBS samples were between 98.3 and 103.6%. The presented method is inexpensive and suitable for clinical testing purposes.</abstract><cop>United States</cop><pub>Oxford University Press</pub><pmid>25344838</pmid><doi>10.1093/chromsci/bmu135</doi><tpages>7</tpages><oa>free_for_read</oa></addata></record> |
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subjects | Adolescent Adult Chromatography, High Pressure Liquid - methods Dried Blood Spot Testing - methods Female Glutathione - blood Glutathione - chemistry Humans Limit of Detection Linear Models Male Methanol Middle Aged Reproducibility of Results Young Adult |
title | Determination of Total Glutathione in Dried Blood Spot Samples Using a High-Performance Liquid Chromatography |
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