Simultaneous determination of 8‐oxo‐2′‐deoxyguanosine and 8‐oxo‐2′‐deoxyadenosine in DNA using online column‐switching liquid chromatography/tandem mass spectrometry

Singh, Rajinder; Teichert, Friederike; Verschoyle, Richard D.; Kaur, Bhavneet; Vives, M.; Sharma, Ricky A.; Steward, William P.; Gescher, Andreas J.; Farmer, Peter B.

doi:10.1002/rcm.3866

Cited by 38 publications

(36 citation statements)

References 41 publications

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“…The reported UPLC–MS/MS method described herein uses TEMPO to prevent artifactual formation or degradation of 8-oxo-dG and is comparable to two other recently published and many other published LC–MS/MS methods. Singh el al [22] recently reported an online column switching LC–MS/MS method for simultaneous quantitation of 8-oxo-dG and 8-oxo-2′-deoxyadenosine in DNA. Endogenous amounts of 8-oxo-dG were different in rat liver DNA isolated with different commercially available DNA isolation kits.…”

Section: Discussionmentioning

confidence: 99%

“…Further, samples can be processed in most contemporary laboratories and sent to a mass spectrometry facility for analysis were subsequent sample analysis is performed in <20 min per sample, making it suitable for application to multi laboratory molecular epidemiology studies. In direct comparison to the other two recent publications, our method does not require column switching technology [22] or custom made antibodies that to our knowledge are not commercially available [17,23]. …”

Section: Discussionmentioning

confidence: 99%

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Analysis of 8-oxo-7,8-dihydro-2′-deoxyguanosine by ultra high pressure liquid chromatography–heat assisted electrospray ionization–tandem mass spectrometry

Boysen

Collins

Liao

et al. 2010

Journal of Chromatography B

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Increased amounts of reactive oxygen species (ROS), generally termed oxidative stress, are frequently hypothesized to be causally associated with many diseases. Analyses of 8-oxo-7,8-dihydro-2′-deoxyguanosine (8-oxo-dG) in DNA and urine are widely used biomarkers for oxidative stress. Over the years it became clear that analysis of 8-oxo-dG in DNA is challenging due to artifactual formation during sample work up. The present study demonstrates that 8-oxo-dG can be measured reliably and accurately when appropriate precautions are taken. First, the presence of an antioxidant, metal chelator, or free radical trapping agent during sample preparation improves reproducibility. Second, sample enrichment by HPLC fraction collection was used to optimize sensitivity. Third, heat assisted electrospray ionization (HESI) eliminated potential interferences and improved assay performance and sensitivity. Subsequently, the UPLC–HESI–MS/MS method was applied to show the biphasic dose response of 8-oxo-dG in H2O2-treated HeLa cells. Application of this method to human lymphocyte DNA (n = 156) gave a mean±SD endogenous amount of 1.57±0.88 adducts per 106 dG, a value that is in agreement with the suggested amount previously estimated by European Standard Committee on Oxidative DNA Damage (ESCODD) and others. These results suggest that the present method is well suited for application to molecular toxicology and epidemiology studies investigating the role of oxidative stress.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Analysis of 8-oxo-7,8-dihydro-2′-deoxyguanosine by ultra high pressure liquid chromatography–heat assisted electrospray ionization–tandem mass spectrometry

Boysen

Collins

Liao

et al. 2010

Journal of Chromatography B

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“…53,55 Increased oxidative DNA adducts were also detected in animal studies exposed to various chemical carcinogens. 39,77–83 Significant increases in HNEdG, M 1 G, and 8-oxo-dG were detected in the liver of rats after an intraperitoneal injection of a single dose of CCl 4 , 77–79 a widely applied model toxicant for hepatic carcinogenesis studies in animals. Trimethylarsine oxide, an organic metabolite of inorganic arsenics, was found to significantly induce the formation of hepatic 8-oxo-dG and hepatocellular adenomas in male F344 rats after two-year exposure with the dose of 200 ppm.…”

Section: Resultsmentioning

confidence: 99%

Polychlorinated Biphenyls Induce Oxidative DNA Adducts in Female Sprague–Dawley Rats

Mutlu

Gao

Collins

et al. 2016

Chem. Res. Toxicol.

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Polychlorinated biphenyls (PCBs) are organic chemicals that were traditionally produced and widely used in industry as mixtures and are presently formed as byproducts of pigment and dye manufacturing. They are known to persist and bioaccumulate in the environment. Some have been shown to induce liver cancer in rodents. Although the mechanism of the toxicity of PCBs is unknown, it has been shown that they increase oxidative stress, including lipid peroxidation. We hypothesized that oxidative stress-induced DNA damage could be a contributor for PCB carcinogenesis and analyzed several DNA adducts in female Sprague–Dawley rats exposed to 3,3′,4,4′,5-pentachlorobiphenyl (PCB 126), 2,2′,4,4′,5,5′-hexachlorobiphenyl (PCB 153), and a binary mixture (PCB 126 + 153) for 14, 31, and 53 wks. Eight adducts were measured to profile oxidative DNA lesions, including 8-oxo-deoxyguanosine (8-oxo-dG), 1,N6-ethenodeoxyadenosine (1,N6-εdA), N2,3-ethenoguanine (N2,3-εG), 1,N2-ethenodeoxyguanosine (1,N2-εdG), as well as malondialdehyde (M1dG), acrolein (AcrdG), crotonaldehyde (CrdG), and 4-hydroxynonenal-derived dG adducts (HNEdG) by LC–MS/MS analysis. Statistically significant increases were observed for 8-oxo-dG and 1,N6-εdA concentrations in hepatic DNA of female rats exposed to the binary mixture (1000 ng/kg/day + 1000 μg/kg/day) but not in rats exposed to PCB 126 (1000 ng/kg/day) or PCB 153 (1000 μg/kg/day) for 14 and 31 wks. However, exposure to PCB 126 (1000 ng/kg/day) for 53 wks significantly increased 8-oxo-dG, 1,N6-εdA, AcrdG, and M1dG. Exposure to PCB 153 (1000 μg/kg/day) for 53 wks increased 8-oxo-dG, and 1,N6-εdA. Exposure to the binary mixture for 53 wks increased 8-oxo-dG, 1,N6-εdA, AcrdG, 1,N2-εdG, and N2,3-εG significantly above control groups. Increased hepatic oxidative DNA adducts following exposure to PCB 126, PCB 153, or the binary mixture shows that an increase in DNA damage may play an important role in hepatic toxicity and carcinogenesis in female Sprague–Dawley rats.

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“…Following elution of the unmodified 2′-deoxynucleosides as well as any matrix impurities that may cause concomitant ionisation suppression, the switching valve diverts the flow from the trap column into the analytical column, which is connected to the mass spectrometer. We have previously applied a similar approach for the determination of DNA adducts derived from reactive oxygen species, acetaldehyde and 3-nitrobenzanthrone [28,34,35]. Interferences from the matrix may result in poor sensitivity by competition for charge, or increased droplet viscosity and surface tension from high concentrations of co-eluting interfering compounds for the formation of gas phase ions in the electrospray source [36].…”

Section: Discussionmentioning

confidence: 99%

Detection and quantitation of N-(deoxyguanosin-8-yl)-2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine adducts in DNA using online column-switching liquid chromatography tandem mass spectrometry

Singh

Arlt

Henderson

et al. 2010

Journal of Chromatography B

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The heterocyclic aromatic amine, 2-amino-1-methyl-6-phenylimidazo [4,5-b]pyridine (PhIP) is formed by the grilled cooking of certain foods such as meats, poultry and fish. PhIP has been shown to induce tumours in the colon, prostate and mammary glands of rats and is regarded as a potential human dietary carcinogen. PhIP is metabolically activated via cytochrome P450 mediated oxidation to an N-hydroxylamino-PhIP intermediate that is subsequently converted to an ester by N-acetyltransferases or sulfotransferases and undergoes heterolytic cleavage to produce a PhIP-nitrenium ion, which reacts with DNA to form the N-(deoxyguanosin-8-yl)-2-amino-1-methyl-6-phenylimidazo [4,5-b]pyridine (PhIP-C8-dG) adduct. Thus far, the detection and quantification of PhIP-DNA adducts has relied to a large extent on 32 P-postlabelling methodologies. In order to expand the array of available techniques for the detection and improved quantification of PhIP-C8-dG adducts in DNA we have developed an online column-switching liquid chromatography (LC)-electrospray ionization (ESI)-tandem mass spectrometry (MS/MS) selected reaction monitoring (SRM) method incorporating an isotopically [ 13 C 10 ]-labelled PhIP-C8-dG internal standard for the analysis of DNA enzymatically hydrolysed to 2′-deoxynucleosides. A dose-dependent increase was observed for PhIP-C8-dG adducts when salmon testis DNA was reacted with N-acetoxy-PhIP. Analysis of DNA samples isolated from colon tissue of mice treated by oral gavage daily for 5 days with 50 mg/kg body weight of PhIP resulted in the detection of an average level of 14.8 ± 3.7 PhIP-C8-dG adducts per 10 6 2′-deoxynucleosides. The method required 50 μg of hydrolysed animal DNA on column and the limit of detection for PhIP-C8-dG was 2.5 fmol (1.5 PhIP-C8-dG adducts per 10 8 2′-deoxynucleosides). In summary, the LC-ESI-MS/MS SRM method provides for the rapid automation of the sample clean up and a reduction in matrix components that would otherwise interfere with the mass spectrometric analysis, with sufficient sensitivity and precision to analyse DNA adducts in animals exposed to PhIP.

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Simultaneous determination of 8‐oxo‐2′‐deoxyguanosine and 8‐oxo‐2′‐deoxyadenosine in DNA using online column‐switching liquid chromatography/tandem mass spectrometry

Cited by 38 publications

References 41 publications

Analysis of 8-oxo-7,8-dihydro-2′-deoxyguanosine by ultra high pressure liquid chromatography–heat assisted electrospray ionization–tandem mass spectrometry

Analysis of 8-oxo-7,8-dihydro-2′-deoxyguanosine by ultra high pressure liquid chromatography–heat assisted electrospray ionization–tandem mass spectrometry

Polychlorinated Biphenyls Induce Oxidative DNA Adducts in Female Sprague–Dawley Rats

Detection and quantitation of N-(deoxyguanosin-8-yl)-2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine adducts in DNA using online column-switching liquid chromatography tandem mass spectrometry

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