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Article: Capillary electrophoresis with LIF detection for assessment of mitochondrial number based on the cardiolipin content

TitleCapillary electrophoresis with LIF detection for assessment of mitochondrial number based on the cardiolipin content
Authors
KeywordsCardiolipin
CE
Laser-induced fluorescence
Mitochondrial number
Issue Date2011
PublisherWiley - V C H Verlag GmbH & Co KGaA.
Citation
Electrophoresis, 2011, v. 32 n. 21, p. 3025-3033 How to Cite?
AbstractCardiolipin is a mitochondria-specific phospholipid known to be intimately involved with numerous mitochondrial functions. Accordingly, quantitative determination of cardiolipin provides a valuable aspect for assessing mitochondrial content and function. The current study was undertaken to develop a simple and reliable method for direct analysis of cardiolipin with particular application for the assessment of mitochondrial number of HepG2 cells. The method presented is based on the online 10-N-nonyl acridine orange (NAO) interaction with cardiolipin using CE with LIF detection. An aqueous-organic solvent system composed of 10% H 2O-40% methanol-50% ACN (all v/v) containing 20μM NAO provides both short analysis time within 2min and a definite fluorescence enhancement at 525nm for NAO-cardiolipin complex as compared with NAO alone. Under the optimum condition, a calibration curve between the peak area and the concentration of cardiolipin was established in the range of 0.1-200μM with a correlation coefficient of 0.9955. The detection limit is 9nM. The proposed method was successfully applied to the analysis of cardiolipin in mitochondria from HepG2 cells. A new biochemical method estimating the mitochondrial number per cell was developed and used together with the proposed method for cardiolipin per cell measurement and cardiolipin per mitochondrion reported before. © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
Persistent Identifierhttp://hdl.handle.net/10722/144515
ISSN
2015 Impact Factor: 2.482
2015 SCImago Journal Rankings: 0.896
ISI Accession Number ID
Funding AgencyGrant Number
Hong Kong Research Grants CouncilHKU 701508
Hong Kong University Research and Conference Grants Committee
Natural Science Foundation of Jiangsu Educational Committee11KJB150018
Priority Academic Program Development of Jiangsu Higher Education Institutions
Funding Information:

The authors would like to acknowledge the financial support from the Hong Kong Research Grants Council (HKU 701508), Seed Funding from the Hong Kong University Research and Conference Grants Committee, the Natural Science Foundation of Jiangsu Educational Committee (No. 11KJB150018) and a Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions.

References

 

DC FieldValueLanguage
dc.contributor.authorZhao, Wen_HK
dc.contributor.authorChen, Qen_HK
dc.contributor.authorWu, Ren_HK
dc.contributor.authorWu, Hen_HK
dc.contributor.authorFung, Yen_HK
dc.contributor.authorO, Wen_HK
dc.date.accessioned2012-02-03T06:11:45Z-
dc.date.available2012-02-03T06:11:45Z-
dc.date.issued2011en_HK
dc.identifier.citationElectrophoresis, 2011, v. 32 n. 21, p. 3025-3033en_HK
dc.identifier.issn0173-0835en_HK
dc.identifier.urihttp://hdl.handle.net/10722/144515-
dc.description.abstractCardiolipin is a mitochondria-specific phospholipid known to be intimately involved with numerous mitochondrial functions. Accordingly, quantitative determination of cardiolipin provides a valuable aspect for assessing mitochondrial content and function. The current study was undertaken to develop a simple and reliable method for direct analysis of cardiolipin with particular application for the assessment of mitochondrial number of HepG2 cells. The method presented is based on the online 10-N-nonyl acridine orange (NAO) interaction with cardiolipin using CE with LIF detection. An aqueous-organic solvent system composed of 10% H 2O-40% methanol-50% ACN (all v/v) containing 20μM NAO provides both short analysis time within 2min and a definite fluorescence enhancement at 525nm for NAO-cardiolipin complex as compared with NAO alone. Under the optimum condition, a calibration curve between the peak area and the concentration of cardiolipin was established in the range of 0.1-200μM with a correlation coefficient of 0.9955. The detection limit is 9nM. The proposed method was successfully applied to the analysis of cardiolipin in mitochondria from HepG2 cells. A new biochemical method estimating the mitochondrial number per cell was developed and used together with the proposed method for cardiolipin per cell measurement and cardiolipin per mitochondrion reported before. © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.en_HK
dc.languageengen_US
dc.publisherWiley - V C H Verlag GmbH & Co KGaA.-
dc.relation.ispartofElectrophoresisen_HK
dc.subjectCardiolipinen_HK
dc.subjectCEen_HK
dc.subjectLaser-induced fluorescenceen_HK
dc.subjectMitochondrial numberen_HK
dc.subject.meshAcetonitriles - chemistryen_HK
dc.subject.meshCardiolipins - analysis - chemistryen_HK
dc.subject.meshElectrophoresis, Capillary - methodsen_HK
dc.subject.meshMitochondria - chemistryen_HK
dc.subject.meshSpectrometry, Fluorescence - methodsen_HK
dc.titleCapillary electrophoresis with LIF detection for assessment of mitochondrial number based on the cardiolipin contenten_HK
dc.typeArticleen_HK
dc.identifier.emailFung, Y:ysfung@hku.hken_HK
dc.identifier.emailO, W:owaisum@hkucc.hku.hken_HK
dc.identifier.authorityFung, Y=rp00697en_HK
dc.identifier.authorityO, W=rp00315en_HK
dc.description.naturelink_to_subscribed_fulltext-
dc.identifier.doi10.1002/elps.201100165en_HK
dc.identifier.pmid22009280-
dc.identifier.scopuseid_2-s2.0-80055023038en_HK
dc.identifier.hkuros198410en_US
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-80055023038&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume32en_HK
dc.identifier.issue21en_HK
dc.identifier.spage3025en_HK
dc.identifier.epage3033en_HK
dc.identifier.isiWOS:000298101000013-
dc.publisher.placeGermanyen_HK
dc.identifier.scopusauthoridZhao, W=34871080300en_HK
dc.identifier.scopusauthoridChen, Q=8060037500en_HK
dc.identifier.scopusauthoridWu, R=54405220300en_HK
dc.identifier.scopusauthoridWu, H=14064321100en_HK
dc.identifier.scopusauthoridFung, Y=13309754700en_HK
dc.identifier.scopusauthoridO, W=6701729369en_HK

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