Detection of DNA Adducts by Electron Capture Mass Spectrometry

Giese, Roger W.

doi:10.1021/tx9601263

Cited by 47 publications

(48 citation statements)

References 53 publications

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“…Our limit of detection is 50 fmol (5×10 −14 mol) of starting material, corresponding to 500 amol (5×10 −16 mol) on the column (data not shown). This compares favorably with the limits of detection observed for other nucleobases analyzed by GC-NCI-MS (27,38), which range from the high zeptomole (zepto = 10 −21 ) to low femtomole (femto = 10 −15 ) levels.…”

Section: Hmdu Can Be Converted Into a Stable Derivative Suitable For supporting

confidence: 64%

Measurement of the Incorporation and Repair of Exogenous 5-Hydroxymethyl-2′-deoxyuridine in Human Cells in Culture Using Gas Chromatography-Negative Chemical Ionization-Mass Spectrometry

Rogstad

Darwanto

Herring

et al. 2007

Chem. Res. Toxicol.

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The DNA of all organisms is constantly damaged by oxidation. Among the array of damage products is 5-hydroxymethyluracil, derived from oxidation of the thymine methyl group. Previous studies have established that HmU can be a sensitive and valuable marker of DNA damage. More recently, the corresponding deoxynucleoside, 5-hydroxymethyl-2′-deoxyuridine (HmdU) has proven to be valuable for the introduction of controlled amounts of a single type of damage lesion into the DNA of replicating cells, which is subsequently repaired by the base excision repair pathway. Complicating the study of HmU formation and repair, however, is the known chemical reactivity of the hydroxymethyl group of HmU under conditions used to hydrolyze DNA. In the work reported here, this chemical property has been exploited by creating conditions that convert HmU to the corresponding methoxymethyluracil (MmU) derivative that can be further derivatized to the 3,5-bis-(trifluoromethyl)benzyl analog. This derivatized compound can be detected by gas chromatographynegative chemical ionization-mass spectrometry (GC-NCI-MS) with good sensitivity. Using isotopically enriched exogenous HmdU and human osteosarcoma cells (U2OS) in culture, we demonstrate that this method allows for the measurement of HmU in DNA formed from incorporation of exogenous HmdU. We further demonstrate that addition of isotopically enriched uridine to the culture medium allows for the simultaneous measurement of DNA replication and repair kinetics. This sensitive and facile method should prove valuable for studies on DNA oxidation damage and repair in living cells.

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Section: Hmdu Can Be Converted Into a Stable Derivative Suitable For supporting

confidence: 64%

Measurement of the Incorporation and Repair of Exogenous 5-Hydroxymethyl-2′-deoxyuridine in Human Cells in Culture Using Gas Chromatography-Negative Chemical Ionization-Mass Spectrometry

Rogstad

Darwanto

Herring

et al. 2007

Chem. Res. Toxicol.

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“…Gases for GC-EC-MS were from MedTech (Medford, MA USA). 2-Fluoro-O 6 -methylhypoxanthine was prepared as described [5].…”

Section: Reagentsmentioning

confidence: 99%

“…The strategy for decreasing noise is to test different electrophoric derivatives until one is found that elutes in a relatively clean region of the GC-EC-MS mass chromatogram. This topic has been discussed in more detail in a review of GC-EC-MS concerned with the detection of DNA adducts [5].…”

Section: Introductionmentioning

confidence: 99%

4-(4′-Methyltetrafluorophenyl)-2,3,5,6-tetrafluorobenzyl bromide: A new electrophoric derivatizing reagent

Murugaiah

Naim

Peng

et al. 2006

Journal of Chromatography A

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Commercially-available 4,4′-dimethyloctafluorobiphenyl was converted in a single step to 4-(4′-methyltetrafluorophenyl)-2,3,5,6-tetrafluorobenzyl bromide (MTFP-TFBBr) for the purpose of providing a new electrophoric derivatizing reagent. When reacted with this reagent, 2-fluoro-O 6 -(2′-hydroxyethyl)hypoxanthine, a model analyte, gave a mixture of isomeric products (apparently substituted at N7 and N9, analogous to its known reaction with pentafluorobenzyl bromide), and 53 femtograms of the mixture was detected at S/N = 10 by gas chromatography electron capture mass spectrometry (GC-EC-MS). As intended, the volatility of the MTFB-TFBBr derivative was much less (2-fold) than that of the corresponding pentafluorobenzyl derivative. It is anticipated that MTFBTFBBr sometimes will be useful in providing an electrophoric derivative that encounters less background noise in analysis by electrophore derivatization/GC-EC-MS.

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“…Gas Chromatography (GC)-MS is less commonly used because many adducts are thermally unstable, and only the DNA adducts that can be chemically derivatized are amenable to GC-MS [65]. Therefore, GC-MS can only be used primarily for specific/limited classes of DNA adducts.…”

Section: Gc-msmentioning

confidence: 99%

“…Metabolic signatures have also been developed for nonhuman primates (NHPs) exposed to external total body irradiation (TBI) [8,38,[64][65][66] and human populations who have received total body irradiation (TBI) as a medical treatment [5]. Identification and quantification of such variations of the metabolites are being investigated for application to biodosimetry and screening of exposed populations by targeted metabolomics.…”

Section: Introductionmentioning

confidence: 99%

Application of liquid chromatography-mass spectrometry and differential mobility spectrometry-mass spectrometry for detection and quantitation of biomarkers in complex biological matrices

Chen¹

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Detection of DNA Adducts by Electron Capture Mass Spectrometry

Cited by 47 publications

References 53 publications

Measurement of the Incorporation and Repair of Exogenous 5-Hydroxymethyl-2′-deoxyuridine in Human Cells in Culture Using Gas Chromatography-Negative Chemical Ionization-Mass Spectrometry

Measurement of the Incorporation and Repair of Exogenous 5-Hydroxymethyl-2′-deoxyuridine in Human Cells in Culture Using Gas Chromatography-Negative Chemical Ionization-Mass Spectrometry

4-(4′-Methyltetrafluorophenyl)-2,3,5,6-tetrafluorobenzyl bromide: A new electrophoric derivatizing reagent

Application of liquid chromatography-mass spectrometry and differential mobility spectrometry-mass spectrometry for detection and quantitation of biomarkers in complex biological matrices

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