Detection and quantification of 8-hydroxy-2′-deoxyguanosine in Alzheimer's transgenic mouse urine using capillary electrophoresis
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作者:
Zhang, Cheng
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Univ Calif San Diego, Sch Med, Dept Neurosci, La Jolla, CA 92093 USA
Louisiana Tech Univ, Dept Biomed Engn, Ruston, LA 71272 USAUniv Calif San Diego, Sch Med, Dept Neurosci, La Jolla, CA 92093 USA
Zhang, Cheng
[1
,2
]
Nestorova, Gergana
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Louisiana Tech Univ, Dept Biomed Engn, Ruston, LA 71272 USAUniv Calif San Diego, Sch Med, Dept Neurosci, La Jolla, CA 92093 USA
Nestorova, Gergana
[2
]
Rissman, Robert A.
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Univ Calif San Diego, Sch Med, Dept Neurosci, La Jolla, CA 92093 USAUniv Calif San Diego, Sch Med, Dept Neurosci, La Jolla, CA 92093 USA
Rissman, Robert A.
[1
]
Feng, June
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Louisiana Tech Univ, Dept Biomed Engn, Ruston, LA 71272 USA
Bossier Parish Community Coll, Div Technol Engn & Math, Bossier City, LA USAUniv Calif San Diego, Sch Med, Dept Neurosci, La Jolla, CA 92093 USA
Feng, June
[2
,3
]
机构:
[1] Univ Calif San Diego, Sch Med, Dept Neurosci, La Jolla, CA 92093 USA
[2] Louisiana Tech Univ, Dept Biomed Engn, Ruston, LA 71272 USA
[3] Bossier Parish Community Coll, Div Technol Engn & Math, Bossier City, LA USA
8-Hydroxy-2'-deoxyguanosine (8-OHdG) is one of the major forms of oxidative DNA damage, and is commonly analyzed as an excellent marker of DNA lesions. The purpose of this study was to develop a sensitive method to accurately and rapidly quantify the 8-OHdG by using CE-LIF detection. The method involved the use of specific antibody to detect the DNA lesion (8-OHdG) and consecutive fluorescence labeling. Next, urinary 8-OHdG fluorescently labeled along with other constituents were resolved by capillary electrophoretic system and the lesion of interest was detected using a fluorescence detector. The limit of detection was 0.18 fmol, which proved sufficient sensitivity for detection and quantification of 8-OHdG in untreated urine samples. The relative standard deviation was found to be 11.32% for migration time and 5.52% for peak area. To demonstrate the utility of this method, the urinary concentration of 8-OHdG in an Alzheimer's transgenic mouse model was determined. Collectively, our results indicate that this methodology offers great advantages, such as high separation efficiency, good selectivity, low limit of detection, simplicity and low cost of analysis.
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CTI, Ctr Tecnol Informacao Renato Archer, Rod D Pedro I,KM 143-6, BR-13069901 Campinas, SP, BrazilCTI, Ctr Tecnol Informacao Renato Archer, Rod D Pedro I,KM 143-6, BR-13069901 Campinas, SP, Brazil
Faria, Aline M.
Peixoto, Elisa B. M. I.
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CTI, Ctr Tecnol Informacao Renato Archer, Rod D Pedro I,KM 143-6, BR-13069901 Campinas, SP, BrazilCTI, Ctr Tecnol Informacao Renato Archer, Rod D Pedro I,KM 143-6, BR-13069901 Campinas, SP, Brazil
Peixoto, Elisa B. M. I.
Adamo, Cristiane B.
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CTI, Ctr Tecnol Informacao Renato Archer, Rod D Pedro I,KM 143-6, BR-13069901 Campinas, SP, BrazilCTI, Ctr Tecnol Informacao Renato Archer, Rod D Pedro I,KM 143-6, BR-13069901 Campinas, SP, Brazil
Adamo, Cristiane B.
Flacker, Alexandre
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CTI, Ctr Tecnol Informacao Renato Archer, Rod D Pedro I,KM 143-6, BR-13069901 Campinas, SP, BrazilCTI, Ctr Tecnol Informacao Renato Archer, Rod D Pedro I,KM 143-6, BR-13069901 Campinas, SP, Brazil
Flacker, Alexandre
Longo, Elson
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Univ Fed Sao Carlos, CDMF, POB 676, BR-13565905 Sao Carlos, SP, BrazilCTI, Ctr Tecnol Informacao Renato Archer, Rod D Pedro I,KM 143-6, BR-13069901 Campinas, SP, Brazil
Longo, Elson
Mazon, Talita
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CTI, Ctr Tecnol Informacao Renato Archer, Rod D Pedro I,KM 143-6, BR-13069901 Campinas, SP, BrazilCTI, Ctr Tecnol Informacao Renato Archer, Rod D Pedro I,KM 143-6, BR-13069901 Campinas, SP, Brazil