Evaluation of the DNA damaging potential of cannabis cigarette smoke by the determination of acetaldehyde derived N2-ethyl-2'-deoxyguanosine adducts.

2.50
Hdl Handle:
http://hdl.handle.net/10146/114460
Title:
Evaluation of the DNA damaging potential of cannabis cigarette smoke by the determination of acetaldehyde derived N2-ethyl-2'-deoxyguanosine adducts.
Authors:
Singh, Rajinder; Sandhu, Jatinderpal; Kaur, Balvinder; Juren, Tina; Steward, William P.; Segerback, Dan; Farmer, Peter B.
Abstract:
Acetaldehyde is an ubiquitous genotoxic compound that has been classified as a possible carcinogen to humans. It can react with DNA to form primarily a Schiff base N(2)-ethylidene-2'-deoxyguanosine (N(2)-ethylidene-dG) adduct. An online column-switching valve liquid chromatography tandem mass spectrometry (LC-MS/MS) selected reaction monitoring (SRM) method was developed for the determination of N(2)-ethylidene-dG adducts in DNA following reduction with sodium cyanoborohydride (NaBH(3)CN) to the chemically stable N(2)-ethyl-2'-deoxyguanosine (N(2)-ethyl-dG) adduct. Accurate quantitation of the adduct was obtained by the addition of the [(15)N(5)]N(2)-ethyl-dG stable isotope-labeled internal standard prior to enzymatic hydrolysis of the DNA samples to 2'-deoxynucleosides with the incorporation of NaBH(3)CN in the DNA hydrolysis buffer. The method required 50 microg of hydrolyzed DNA on column for the analysis, and the limit of detection for N(2)-ethyl-dG was 2.0 fmol. The analysis of calf thymus DNA treated in vitro with acetaldehyde (ranging from 0.5 to 100 mM) or with the smoke generated from 1, 5, and 10 cannabis cigarettes showed linear dose-dependent increases in the level of N(2)-ethyl-dG adducts (r = 0.954 and r = 0.999, respectively). Similar levels (332.8 +/- 21.9 vs 348.4 +/- 19.1 adducts per 10(8) 2'-deoxynucleosides) of N(2)-ethyl-dG adducts were detected following the exposure of calf thymus DNA to 10 tobacco or 10 cannabis cigarettes. No significant difference was found in the levels of N(2)-ethyl-dG adducts in human lung DNA obtained from nonsmokers (n = 4) and smokers (n = 4) with the average level observed as 13.3 +/- 0.7 adducts per 10(8) 2'-deoxynucleosides. No N(2)-ethyl-dG adducts were detected in any of the DNA samples following analysis with the omission of NaBH(3)CN from the DNA hydrolysis buffer. In conclusion, these results provide evidence for the DNA damaging potential of cannabis smoke, implying that the consumption of cannabis cigarettes may be detrimental to human health with the possibility to initiate cancer development.
Citation:
Chem. Res. Toxicol. 2009, 22 (6):1181-1188
Journal:
Chemical Research in Toxicology
Issue Date:
Jun-2009
URI:
http://hdl.handle.net/10146/114460
DOI:
10.1021/tx900106y
PubMed ID:
19449825
Additional Links:
http://pubs.acs.org/doi/full/10.1021/tx900106y
Type:
Article
Language:
en
ISSN:
1520-5010
Sponsors:
We acknowledge the following financial support: the Environmental Cancer Risk, Nutrition and Individual Susceptibility, a European Union Network of Excellence (ECNIS, Contract No. FOOD-CT-2005-513943), ECNIS Type B project
Appears in Collections:
Articles

Full metadata record

DC FieldValue Language
dc.contributor.authorSingh, Rajinderen
dc.contributor.authorSandhu, Jatinderpalen
dc.contributor.authorKaur, Balvinderen
dc.contributor.authorJuren, Tinaen
dc.contributor.authorSteward, William P.en
dc.contributor.authorSegerback, Danen
dc.contributor.authorFarmer, Peter B.en
dc.date.accessioned2010-11-03T11:46:38Z-
dc.date.available2010-11-03T11:46:38Z-
dc.date.issued2009-06-
dc.identifier.citationChem. Res. Toxicol. 2009, 22 (6):1181-1188en
dc.identifier.issn1520-5010-
dc.identifier.pmid19449825-
dc.identifier.doi10.1021/tx900106y-
dc.identifier.urihttp://hdl.handle.net/10146/114460-
dc.description.abstractAcetaldehyde is an ubiquitous genotoxic compound that has been classified as a possible carcinogen to humans. It can react with DNA to form primarily a Schiff base N(2)-ethylidene-2'-deoxyguanosine (N(2)-ethylidene-dG) adduct. An online column-switching valve liquid chromatography tandem mass spectrometry (LC-MS/MS) selected reaction monitoring (SRM) method was developed for the determination of N(2)-ethylidene-dG adducts in DNA following reduction with sodium cyanoborohydride (NaBH(3)CN) to the chemically stable N(2)-ethyl-2'-deoxyguanosine (N(2)-ethyl-dG) adduct. Accurate quantitation of the adduct was obtained by the addition of the [(15)N(5)]N(2)-ethyl-dG stable isotope-labeled internal standard prior to enzymatic hydrolysis of the DNA samples to 2'-deoxynucleosides with the incorporation of NaBH(3)CN in the DNA hydrolysis buffer. The method required 50 microg of hydrolyzed DNA on column for the analysis, and the limit of detection for N(2)-ethyl-dG was 2.0 fmol. The analysis of calf thymus DNA treated in vitro with acetaldehyde (ranging from 0.5 to 100 mM) or with the smoke generated from 1, 5, and 10 cannabis cigarettes showed linear dose-dependent increases in the level of N(2)-ethyl-dG adducts (r = 0.954 and r = 0.999, respectively). Similar levels (332.8 +/- 21.9 vs 348.4 +/- 19.1 adducts per 10(8) 2'-deoxynucleosides) of N(2)-ethyl-dG adducts were detected following the exposure of calf thymus DNA to 10 tobacco or 10 cannabis cigarettes. No significant difference was found in the levels of N(2)-ethyl-dG adducts in human lung DNA obtained from nonsmokers (n = 4) and smokers (n = 4) with the average level observed as 13.3 +/- 0.7 adducts per 10(8) 2'-deoxynucleosides. No N(2)-ethyl-dG adducts were detected in any of the DNA samples following analysis with the omission of NaBH(3)CN from the DNA hydrolysis buffer. In conclusion, these results provide evidence for the DNA damaging potential of cannabis smoke, implying that the consumption of cannabis cigarettes may be detrimental to human health with the possibility to initiate cancer development.en
dc.description.sponsorshipWe acknowledge the following financial support: the Environmental Cancer Risk, Nutrition and Individual Susceptibility, a European Union Network of Excellence (ECNIS, Contract No. FOOD-CT-2005-513943), ECNIS Type B projecten
dc.language.isoenen
dc.relation.urlhttp://pubs.acs.org/doi/full/10.1021/tx900106yen
dc.subjectCannabisen
dc.subjectCarcinogensen
dc.subjectMarijuana Smokingen
dc.subjectDNA Adductsen
dc.subjectDNA Damageen
dc.subjectAcetaldehydeen
dc.subjectDeoxyguanosineen
dc.subjectHumansen
dc.subject.meshAcetaldehyde-
dc.subject.meshAdult-
dc.subject.meshCannabis-
dc.subject.meshCarcinogens-
dc.subject.meshChromatography, High Pressure Liquid-
dc.subject.meshDNA-
dc.subject.meshDNA Adducts-
dc.subject.meshDNA Damage-
dc.subject.meshDeoxyguanosine-
dc.subject.meshHumans-
dc.subject.meshLung-
dc.subject.meshMale-
dc.subject.meshMarijuana Smoking-
dc.subject.meshMiddle Aged-
dc.subject.meshSpectrometry, Mass, Electrospray Ionization-
dc.titleEvaluation of the DNA damaging potential of cannabis cigarette smoke by the determination of acetaldehyde derived N2-ethyl-2'-deoxyguanosine adducts.en
dc.typeArticleen
dc.identifier.journalChemical Research in Toxicologyen

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