Volume 20 Issue 5
Oct.  2007
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JIANG YUE, PENG WANG, YING-HUI LIU, JUN-YU WU, JIE CHEN, REN-XIU PENG. Fast Evaluation of Oxidative DNA Damage by Liquid Chromatography-Electropray Tandem Mass Spectrometry Coupled With Precision-Cut Rat Liver Slices[J]. Biomedical and Environmental Sciences, 2007, 20(5): 386-391.
Citation: JIANG YUE, PENG WANG, YING-HUI LIU, JUN-YU WU, JIE CHEN, REN-XIU PENG. Fast Evaluation of Oxidative DNA Damage by Liquid Chromatography-Electropray Tandem Mass Spectrometry Coupled With Precision-Cut Rat Liver Slices[J]. Biomedical and Environmental Sciences, 2007, 20(5): 386-391.

Fast Evaluation of Oxidative DNA Damage by Liquid Chromatography-Electropray Tandem Mass Spectrometry Coupled With Precision-Cut Rat Liver Slices

Funds:  国家自然科学基金(30600773)
  • 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. Methods Precision-cut liver slices (300 μm) 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℃. 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). Results 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). Conclusion 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.
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    沈阳化工大学材料科学与工程学院 沈阳 110142

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Fast Evaluation of Oxidative DNA Damage by Liquid Chromatography-Electropray Tandem Mass Spectrometry Coupled With Precision-Cut Rat Liver Slices

Funds:  国家自然科学基金(30600773)

Abstract: 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. Methods Precision-cut liver slices (300 μm) 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℃. 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). Results 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). Conclusion 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.

JIANG YUE, PENG WANG, YING-HUI LIU, JUN-YU WU, JIE CHEN, REN-XIU PENG. Fast Evaluation of Oxidative DNA Damage by Liquid Chromatography-Electropray Tandem Mass Spectrometry Coupled With Precision-Cut Rat Liver Slices[J]. Biomedical and Environmental Sciences, 2007, 20(5): 386-391.
Citation: JIANG YUE, PENG WANG, YING-HUI LIU, JUN-YU WU, JIE CHEN, REN-XIU PENG. Fast Evaluation of Oxidative DNA Damage by Liquid Chromatography-Electropray Tandem Mass Spectrometry Coupled With Precision-Cut Rat Liver Slices[J]. Biomedical and Environmental Sciences, 2007, 20(5): 386-391.

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