Development of a quantitative liquid chromatography/electrospray mass spectrometric assay for a mutagenic tobacco specific nitrosamine-derived DNA adduct, O6-[4-oxo-4-(3-pyridyl)butyl]-2′-deoxyguanosine

Nicole M. Thomson, Renée S. Mijal, Rebecca Ziegel, Nancy L. Fleischer, Anthony E. Pegg, Natalia Y. Tretyakova, Lisa A. Peterson

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

Liquid chromatography with electrospray tandem mass spectrometry (LC/ESI-MS/MS) was employed to quantify O6-[4-oxo-4-(3-pyridyl)butyl] -2′-deoxyguanosine (O6-pobdG), a mutagenic adduct formed by pyridyloxobutylating nitrosamines. Selected reaction monitoring (SRM) of the neutral loss of the sugar from protonated molecules of the adduct, [M + H - 116]+, was utilized for detection of O6-pobdG in pyridyloxobutylated DNA from both in vitro and in vivo sources. Quantitation was based on isotope dilution with synthetic O6-[1,2,2- 2H3-4-oxo-4-(3-pyridyl)butyl]-2′-deoxyguanosine. The detection limits in this study were less than 5 fmol of pure standard and 50 fmol in 1.5 mg of DNA. This method was validated by comparing adduct levels measured with the LC/ESI-MS/MS method to those obtained with radiochemical methods in DNA alkylated with the model pyridyloxobutylating agent, [5- 3H]4-(acetoxymethylnitrosamino)-1-(3-pyridyl)-1-butanone ([5- 3H]NNKOAc). The pyridyloxobutyl 2′-deoxyguanosine adduct coeluting with the deuterated standard disappeared when NNKOAc-treated DNA had been reacted with the repair protein, O6-alkylguanine-DNA alkyltransferase. This result confirms that the coeluting peak is solely O 6-pobdG. Preliminary studies with liver DNA isolated from NNKOAc-treated mice demonstrated that this method can be used to quantify O 6-pobG in DNA from in vivo sources. The improved sensitivity and specificity of adduct detection afforded by this LC/ESI-MS/MS method will allow us to explore the role of O6-pobdG in the toxicological properties of pyridyloxobutylating nitrosamines.

Original languageEnglish (US)
Pages (from-to)1600-1606
Number of pages7
JournalChemical research in toxicology
Volume17
Issue number12
DOIs
StatePublished - Dec 1 2004

Fingerprint

Nitrosamines
Tobacco
DNA Adducts
Liquid chromatography
Liquid Chromatography
Assays
DNA
Deoxyguanosine
Tandem Mass Spectrometry
Sugars
Isotopes
Liver
Toxicology
Dilution
Mass spectrometry
Limit of Detection
O(6)-(4-oxo-4-(3-pyridyl)butyl)guanine
Repair
Sensitivity and Specificity
Molecules

All Science Journal Classification (ASJC) codes

  • Toxicology

Cite this

Thomson, Nicole M. ; Mijal, Renée S. ; Ziegel, Rebecca ; Fleischer, Nancy L. ; Pegg, Anthony E. ; Tretyakova, Natalia Y. ; Peterson, Lisa A. / Development of a quantitative liquid chromatography/electrospray mass spectrometric assay for a mutagenic tobacco specific nitrosamine-derived DNA adduct, O6-[4-oxo-4-(3-pyridyl)butyl]-2′-deoxyguanosine. In: Chemical research in toxicology. 2004 ; Vol. 17, No. 12. pp. 1600-1606.
@article{976ce8f84a274ce29b99eb4ffe3363a4,
title = "Development of a quantitative liquid chromatography/electrospray mass spectrometric assay for a mutagenic tobacco specific nitrosamine-derived DNA adduct, O6-[4-oxo-4-(3-pyridyl)butyl]-2′-deoxyguanosine",
abstract = "Liquid chromatography with electrospray tandem mass spectrometry (LC/ESI-MS/MS) was employed to quantify O6-[4-oxo-4-(3-pyridyl)butyl] -2′-deoxyguanosine (O6-pobdG), a mutagenic adduct formed by pyridyloxobutylating nitrosamines. Selected reaction monitoring (SRM) of the neutral loss of the sugar from protonated molecules of the adduct, [M + H - 116]+, was utilized for detection of O6-pobdG in pyridyloxobutylated DNA from both in vitro and in vivo sources. Quantitation was based on isotope dilution with synthetic O6-[1,2,2- 2H3-4-oxo-4-(3-pyridyl)butyl]-2′-deoxyguanosine. The detection limits in this study were less than 5 fmol of pure standard and 50 fmol in 1.5 mg of DNA. This method was validated by comparing adduct levels measured with the LC/ESI-MS/MS method to those obtained with radiochemical methods in DNA alkylated with the model pyridyloxobutylating agent, [5- 3H]4-(acetoxymethylnitrosamino)-1-(3-pyridyl)-1-butanone ([5- 3H]NNKOAc). The pyridyloxobutyl 2′-deoxyguanosine adduct coeluting with the deuterated standard disappeared when NNKOAc-treated DNA had been reacted with the repair protein, O6-alkylguanine-DNA alkyltransferase. This result confirms that the coeluting peak is solely O 6-pobdG. Preliminary studies with liver DNA isolated from NNKOAc-treated mice demonstrated that this method can be used to quantify O 6-pobG in DNA from in vivo sources. The improved sensitivity and specificity of adduct detection afforded by this LC/ESI-MS/MS method will allow us to explore the role of O6-pobdG in the toxicological properties of pyridyloxobutylating nitrosamines.",
author = "Thomson, {Nicole M.} and Mijal, {Ren{\'e}e S.} and Rebecca Ziegel and Fleischer, {Nancy L.} and Pegg, {Anthony E.} and Tretyakova, {Natalia Y.} and Peterson, {Lisa A.}",
year = "2004",
month = "12",
day = "1",
doi = "10.1021/tx0498298",
language = "English (US)",
volume = "17",
pages = "1600--1606",
journal = "Chemical Research in Toxicology",
issn = "0893-228X",
publisher = "American Chemical Society",
number = "12",

}

Development of a quantitative liquid chromatography/electrospray mass spectrometric assay for a mutagenic tobacco specific nitrosamine-derived DNA adduct, O6-[4-oxo-4-(3-pyridyl)butyl]-2′-deoxyguanosine. / Thomson, Nicole M.; Mijal, Renée S.; Ziegel, Rebecca; Fleischer, Nancy L.; Pegg, Anthony E.; Tretyakova, Natalia Y.; Peterson, Lisa A.

In: Chemical research in toxicology, Vol. 17, No. 12, 01.12.2004, p. 1600-1606.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Development of a quantitative liquid chromatography/electrospray mass spectrometric assay for a mutagenic tobacco specific nitrosamine-derived DNA adduct, O6-[4-oxo-4-(3-pyridyl)butyl]-2′-deoxyguanosine

AU - Thomson, Nicole M.

AU - Mijal, Renée S.

AU - Ziegel, Rebecca

AU - Fleischer, Nancy L.

AU - Pegg, Anthony E.

AU - Tretyakova, Natalia Y.

AU - Peterson, Lisa A.

PY - 2004/12/1

Y1 - 2004/12/1

N2 - Liquid chromatography with electrospray tandem mass spectrometry (LC/ESI-MS/MS) was employed to quantify O6-[4-oxo-4-(3-pyridyl)butyl] -2′-deoxyguanosine (O6-pobdG), a mutagenic adduct formed by pyridyloxobutylating nitrosamines. Selected reaction monitoring (SRM) of the neutral loss of the sugar from protonated molecules of the adduct, [M + H - 116]+, was utilized for detection of O6-pobdG in pyridyloxobutylated DNA from both in vitro and in vivo sources. Quantitation was based on isotope dilution with synthetic O6-[1,2,2- 2H3-4-oxo-4-(3-pyridyl)butyl]-2′-deoxyguanosine. The detection limits in this study were less than 5 fmol of pure standard and 50 fmol in 1.5 mg of DNA. This method was validated by comparing adduct levels measured with the LC/ESI-MS/MS method to those obtained with radiochemical methods in DNA alkylated with the model pyridyloxobutylating agent, [5- 3H]4-(acetoxymethylnitrosamino)-1-(3-pyridyl)-1-butanone ([5- 3H]NNKOAc). The pyridyloxobutyl 2′-deoxyguanosine adduct coeluting with the deuterated standard disappeared when NNKOAc-treated DNA had been reacted with the repair protein, O6-alkylguanine-DNA alkyltransferase. This result confirms that the coeluting peak is solely O 6-pobdG. Preliminary studies with liver DNA isolated from NNKOAc-treated mice demonstrated that this method can be used to quantify O 6-pobG in DNA from in vivo sources. The improved sensitivity and specificity of adduct detection afforded by this LC/ESI-MS/MS method will allow us to explore the role of O6-pobdG in the toxicological properties of pyridyloxobutylating nitrosamines.

AB - Liquid chromatography with electrospray tandem mass spectrometry (LC/ESI-MS/MS) was employed to quantify O6-[4-oxo-4-(3-pyridyl)butyl] -2′-deoxyguanosine (O6-pobdG), a mutagenic adduct formed by pyridyloxobutylating nitrosamines. Selected reaction monitoring (SRM) of the neutral loss of the sugar from protonated molecules of the adduct, [M + H - 116]+, was utilized for detection of O6-pobdG in pyridyloxobutylated DNA from both in vitro and in vivo sources. Quantitation was based on isotope dilution with synthetic O6-[1,2,2- 2H3-4-oxo-4-(3-pyridyl)butyl]-2′-deoxyguanosine. The detection limits in this study were less than 5 fmol of pure standard and 50 fmol in 1.5 mg of DNA. This method was validated by comparing adduct levels measured with the LC/ESI-MS/MS method to those obtained with radiochemical methods in DNA alkylated with the model pyridyloxobutylating agent, [5- 3H]4-(acetoxymethylnitrosamino)-1-(3-pyridyl)-1-butanone ([5- 3H]NNKOAc). The pyridyloxobutyl 2′-deoxyguanosine adduct coeluting with the deuterated standard disappeared when NNKOAc-treated DNA had been reacted with the repair protein, O6-alkylguanine-DNA alkyltransferase. This result confirms that the coeluting peak is solely O 6-pobdG. Preliminary studies with liver DNA isolated from NNKOAc-treated mice demonstrated that this method can be used to quantify O 6-pobG in DNA from in vivo sources. The improved sensitivity and specificity of adduct detection afforded by this LC/ESI-MS/MS method will allow us to explore the role of O6-pobdG in the toxicological properties of pyridyloxobutylating nitrosamines.

UR - http://www.scopus.com/inward/record.url?scp=10844279953&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=10844279953&partnerID=8YFLogxK

U2 - 10.1021/tx0498298

DO - 10.1021/tx0498298

M3 - Article

C2 - 15606135

AN - SCOPUS:10844279953

VL - 17

SP - 1600

EP - 1606

JO - Chemical Research in Toxicology

JF - Chemical Research in Toxicology

SN - 0893-228X

IS - 12

ER -