Fast evaluation of oxidative DNA damage by liquid chromatography-electrospray tandem mass spectrometry coupled with precision-cut rat liver slices / 生物医学与环境科学(英文)
Biomedical and Environmental Sciences
;
(12): 386-391, 2007.
Article
in English
| WPRIM
| ID: wpr-249838
ABSTRACT
<p><b>OBJECTIVE</b>To establish a fast and sensitive method for the detection of 8-hydroxy-2'-deoxyguanosine (8-OHdG) in precision-cut rat liver slices by HPLC-MS/MS and to investigate isoniazid (INH) -induced oxidative DNA damage.</p><p><b>METHODS</b>Precision-cut liver slices (300 microm) were prepared from male rats, and incubated with INH (0.018 mol/L) for 2 h after 1 h preincubation. DNA in the slices was extracted and digested into free nucleosides at 37 degrees C. The samples were injected into HPLC-MS/MS after the proteins were removed. The level of oxidative DNA damage was estimated using the ratio of 8-OHdG to deoxyguanosine (dG).</p><p><b>RESULTS</b>The limit of detection of 8-OHdG was 1 ng/mL (S/N=3) and the intra-assay relative standard variation was 3.38% when one transition 284.3/168.4 was used as a quantifier and another two transitions 284.3/140.2, 306.1/190.2 as qualifiers. 8-OHdG and dG were well separated, as indicated by elution at 10.02 and 7.37 min, respectively. INH significantly increased the ratio of 8-OHdG to dG in rat liver slices (P<0.05).</p><p><b>CONCLUSION</b>8-OHdG in precision-cut liver slices could be sensitively determined by HPLC-MS/MS. HPLC-MS/MS coupled with precision-cut tissue slices is a fast and reliable analytical technique to evaluate oxidative DNA damage of target tissues caused by procarcinogens and cytotoxins.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Pharmacology
/
Time Factors
/
DNA Damage
/
Chromatography, Liquid
/
Sensitivity and Specificity
/
Rats, Wistar
/
Oxidative Stress
/
Spectrometry, Mass, Electrospray Ionization
/
Deoxyguanosine
/
Isoniazid
Type of study:
Diagnostic study
Limits:
Animals
/
Humans
/
Male
Language:
English
Journal:
Biomedical and Environmental Sciences
Year:
2007
Type:
Article
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