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Article: Rapid preparation of human urine and plasma samples for analysis of F 2-isoprostanes by gas chromatography-mass spectrometry

TitleRapid preparation of human urine and plasma samples for analysis of F 2-isoprostanes by gas chromatography-mass spectrometry
Authors
Issue Date2004
PublisherAcademic Press. The Journal's web site is located at http://www.elsevier.com/wps/find/journaldescription.cws_home/622790/description
Citation
Biochemical And Biophysical Research Communications, 2004, v. 320 n. 3, p. 696-702 How to Cite?
AbstractReliable MS-based methods have been developed for the measurement of free and esterified F2-isoprostanes. However, prior to sample analysis several steps of purification, including solid-phase extraction followed by TLC or HPLC, are usually required, making it tedious to analyze large sample numbers, e.g., for population studies. We report a quick sample purification method using anion exchange solid phase extraction (SPE), which is highly selective for acidic compounds. Urine and hydrolyzed plasma of healthy individuals were acidified before SPE extraction, washed with 4 different solvent mixtures and finally eluted with ethyl acetate. The eluted samples were first derivatized with pentafluorobenzyl bromide followed by a second derivatization with bis-(trimethylsilyl)trifluoroacetamide. F 2-isoprostanes were analyzed by GC-MS-NCI. The method was highly sensitive; the limit of detection at 5:1 signal-to-noise ratio was 0.037ng/ml and 0.007ng/mg creatinine for plasma and urine, respectively. Anion exchange SPE extraction for F2-isoprostane showed recovery of 55-65% and high linearity for concentration 0-1.0ng/ml for urine (CV=4.08%, r2=0.990) and 0-0.5ng/ml for plasma (CV=4.07%, r2=0.998). Fasting for 6h significantly increased plasma F2-isoprostanes levels, which has implications for the design of intervention studies using this biomarker. © 2004 Elsevier Inc. All rights reserved.
Persistent Identifierhttp://hdl.handle.net/10722/178925
ISSN
2015 Impact Factor: 2.371
2015 SCImago Journal Rankings: 1.152
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorLee, CYJen_US
dc.contributor.authorJenner, AMen_US
dc.contributor.authorHalliwell, Ben_US
dc.date.accessioned2012-12-19T09:50:46Z-
dc.date.available2012-12-19T09:50:46Z-
dc.date.issued2004en_US
dc.identifier.citationBiochemical And Biophysical Research Communications, 2004, v. 320 n. 3, p. 696-702en_US
dc.identifier.issn0006-291Xen_US
dc.identifier.urihttp://hdl.handle.net/10722/178925-
dc.description.abstractReliable MS-based methods have been developed for the measurement of free and esterified F2-isoprostanes. However, prior to sample analysis several steps of purification, including solid-phase extraction followed by TLC or HPLC, are usually required, making it tedious to analyze large sample numbers, e.g., for population studies. We report a quick sample purification method using anion exchange solid phase extraction (SPE), which is highly selective for acidic compounds. Urine and hydrolyzed plasma of healthy individuals were acidified before SPE extraction, washed with 4 different solvent mixtures and finally eluted with ethyl acetate. The eluted samples were first derivatized with pentafluorobenzyl bromide followed by a second derivatization with bis-(trimethylsilyl)trifluoroacetamide. F 2-isoprostanes were analyzed by GC-MS-NCI. The method was highly sensitive; the limit of detection at 5:1 signal-to-noise ratio was 0.037ng/ml and 0.007ng/mg creatinine for plasma and urine, respectively. Anion exchange SPE extraction for F2-isoprostane showed recovery of 55-65% and high linearity for concentration 0-1.0ng/ml for urine (CV=4.08%, r2=0.990) and 0-0.5ng/ml for plasma (CV=4.07%, r2=0.998). Fasting for 6h significantly increased plasma F2-isoprostanes levels, which has implications for the design of intervention studies using this biomarker. © 2004 Elsevier Inc. All rights reserved.en_US
dc.languageengen_US
dc.publisherAcademic Press. The Journal's web site is located at http://www.elsevier.com/wps/find/journaldescription.cws_home/622790/descriptionen_US
dc.relation.ispartofBiochemical and Biophysical Research Communicationsen_US
dc.subject.meshAdulten_US
dc.subject.meshBlood Chemical Analysis - Methodsen_US
dc.subject.meshF2-Isoprostanes - Blood - Urineen_US
dc.subject.meshFemaleen_US
dc.subject.meshGas Chromatography-Mass Spectrometry - Methodsen_US
dc.subject.meshHumansen_US
dc.subject.meshMaleen_US
dc.subject.meshReproducibility Of Resultsen_US
dc.subject.meshSensitivity And Specificityen_US
dc.subject.meshSpecimen Handling - Methodsen_US
dc.subject.meshUrinalysis - Methodsen_US
dc.titleRapid preparation of human urine and plasma samples for analysis of F 2-isoprostanes by gas chromatography-mass spectrometryen_US
dc.typeArticleen_US
dc.identifier.emailLee, CYJ: jettylee@hku.hken_US
dc.identifier.authorityLee, CYJ=rp01511en_US
dc.description.naturelink_to_subscribed_fulltexten_US
dc.identifier.doi10.1016/j.bbrc.2004.06.015en_US
dc.identifier.pmid15240104-
dc.identifier.scopuseid_2-s2.0-3142528536en_US
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-3142528536&selection=ref&src=s&origin=recordpageen_US
dc.identifier.volume320en_US
dc.identifier.issue3en_US
dc.identifier.spage696en_US
dc.identifier.epage702en_US
dc.identifier.isiWOS:000222723200011-
dc.publisher.placeUnited Statesen_US
dc.identifier.scopusauthoridLee, CYJ=13104265200en_US
dc.identifier.scopusauthoridJenner, AM=7006757910en_US
dc.identifier.scopusauthoridHalliwell, B=7101878919en_US

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