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.

2.50
Hdl Handle:
http://hdl.handle.net/10146/192390
Title:
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.
Authors:
Singh, Rajinder; Arlt, Volker M.; Henderson, Colin J.; Phillips, David H.; Farmer, Peter B.; Gamboa da Costa, Goncalo
Abstract:
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 microg 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.
Citation:
J. Chromatogr. B Analyt. Technol. Biomed. Life Sci. 2010, 878 (23):2155-2162
Journal:
Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
Issue Date:
1-Aug-2010
URI:
http://hdl.handle.net/10146/192390
DOI:
10.1016/j.jchromb.2010.06.008
PubMed ID:
20598652
Additional Links:
http://www.sciencedirect.com/science/article/pii/S1570023210003739; http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2923026/?tool=pubmed
Type:
Article
Language:
en
ISSN:
1873-376X
Sponsors:
The authors acknowledge financial support from the European Union Network of Excellence ECNIS, Type B project (Environmental Cancer Risk, Nutrition and Individual Susceptibility, www.ecnis.org, Contract No. FOOD-CT-2005-513943)
Appears in Collections:
Articles

Full metadata record

DC FieldValue Language
dc.contributor.authorSingh, Rajinderen
dc.contributor.authorArlt, Volker M.en
dc.contributor.authorHenderson, Colin J.en
dc.contributor.authorPhillips, David H.en
dc.contributor.authorFarmer, Peter B.en
dc.contributor.authorGamboa da Costa, Goncaloen
dc.date.accessioned2011-11-29T14:13:07Z-
dc.date.available2011-11-29T14:13:07Z-
dc.date.issued2010-08-01-
dc.identifier.citationJ. Chromatogr. B Analyt. Technol. Biomed. Life Sci. 2010, 878 (23):2155-2162en
dc.identifier.issn1873-376X-
dc.identifier.pmid20598652-
dc.identifier.doi10.1016/j.jchromb.2010.06.008-
dc.identifier.urihttp://hdl.handle.net/10146/192390-
dc.description.abstractThe 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 microg 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.en
dc.description.sponsorshipThe authors acknowledge financial support from the European Union Network of Excellence ECNIS, Type B project (Environmental Cancer Risk, Nutrition and Individual Susceptibility, www.ecnis.org, Contract No. FOOD-CT-2005-513943)en
dc.language.isoenen
dc.relation.urlhttp://www.sciencedirect.com/science/article/pii/S1570023210003739en
dc.relation.urlhttp://www.ncbi.nlm.nih.gov/pmc/articles/PMC2923026/?tool=pubmeden
dc.subject2-Amino-1-methyl-6-phenylimidazo[4,5-b]pyridine(PhIP)en
dc.subjectDNA adductsen
dc.subjectLC–MS/MSen
dc.subjectChemical carcinogenen
dc.subject.meshAnimals-
dc.subject.meshCalibration-
dc.subject.meshChromatography, Liquid-
dc.subject.meshDNA-
dc.subject.meshDNA Adducts-
dc.subject.meshDeoxyguanosine-
dc.subject.meshImidazoles-
dc.subject.meshMice-
dc.subject.meshOnline Systems-
dc.subject.meshReproducibility of Results-
dc.subject.meshSpectrometry, Mass, Electrospray Ionization-
dc.subject.meshTandem Mass Spectrometry-
dc.titleDetection 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.en
dc.typeArticleen
dc.identifier.journalJournal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciencesen

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