Home LiteratureArticle Details
PMID: 17305409 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Novel LC-ESI/MS/MS(n) method for the characterization and quantification of 2'-deoxyguanosine adducts of the dietary carcinogen 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine by 2-D linear quadrupole ion trap mass spectrometry.

Chemical research in toxicology ·Vol. 20 ·No. 2 ·2007-02-00 ·Pages 263-76

Goodenough AK, Schut HA, Turesky RJ

Abstract

An accurate and sensitive liquid chromatography-electrospray ionization/multi-stage mass spectrometry (LC-ESI/MS/MS(n)) technique has been developed for the characterization and quantification of 2'-deoxyguanosine (dG) adducts of the dietary mutagen, 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP). PhIP is an animal and potential human carcinogen that occurs in grilled meats. Following enzymatic digestion and adduct enrichment by solid-phase extraction (SPE), PhIP-DNA adducts were analyzed by MS/MS and MS(n) scan modes on a 2-D linear quadrupole ion trap mass spectrometer (QIT/MS). The major DNA adduct, N-(deoxyguanosin-8-yl)-2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (dG-C8-PhIP), was detected in calf thymus (CT) DNA modified in vitro with a bioactivated form of PhIP and in the colon and liver of rats given PhIP as part of the diet. The lower limit of detection (LOD) was 1 adduct per 10(8) DNA bases, and the limit of quantification (LOQ) was 3 adducts per 10(8) DNA bases in both MS/MS and MS(3) scan modes, using 27 microg of DNA for analysis. Measurements were based on isotope dilution with the internal standard, N-(deoxyguanosin-8-yl)-2-amino-1-(trideutero)methyl-6-phenylimidazo[4,5-b]pyridine (dG-C8-[2H3C]-PhIP). The selected reaction monitoring (SRM) scan mode in MS/MS was employed to monitor the loss of deoxyribose (dR) from the protonated molecules of the adducts ([M + H - 116]+). The consecutive reaction monitoring (CRM) scan modes in MS(3) and MS(4) were used to measure and further characterize product ions of the aglycone ion (BH2+) (Guanyl-PhIP). The MS(3) scan mode was effective in eliminating isobaric interferences observed in the MS/MS scan mode and resulted in an improved signal-to-noise (S/N) ratio. Moreover, the product ion spectra obtained by the MS(n) scan modes provided rich structural information about the adduct and were used to corroborate the identity of dG-C8-PhIP. In addition, an isomeric dG-PhIP adduct was detected in vivo. This LC-ESI/MS/MS(n) method is the first reported application on the use of the MS(3) scan mode for the analysis of DNA adducts in vivo.

MeSH Terms
Animals Carcinogens/administration & dosage,chemistry,metabolism Chromatography, Liquid/methods Colon/chemistry,drug effects,metabolism DNA Adducts/analysis,metabolism Deoxyguanosine/analogs & derivatives,analysis,metabolism Diet Female Imidazoles/administration & dosage,analysis,toxicity Kinetics Liver/chemistry,drug effects,metabolism Molecular Structure Rats Rats, Sprague-Dawley Sensitivity and Specificity Spectrometry, Mass, Electrospray Ionization/methods Stereoisomerism Tandem Mass Spectrometry/methods Time Factors
Chemicals
Carcinogens DNA Adducts Imidazoles 2-amino-1-methyl-6-phenylimidazo(4,5-b)pyridine Deoxyguanosine
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Goodenough Angela K
Division of Environmental Disease Prevention, Wadsworth Center, NYS Department of Health, Albany, New York 12201-0509, USA.
Schut Herman A J
Turesky Robert J
References (68)
68 references, click to expand
  1. DNA adducts: biological markers of exposure and potential applications to risk assessment.
    Mutat Res. 1996 Sep;365(1-3):129-46 PMID: 8898994
  2. Mechanism of C8 alkylation of guanine residues by activated arylamines: evidence for initial adduct formation at the N7 position.
    Proc Natl Acad Sci U S A. 1992 Sep 1;89(17):8278-82 PMID: 1518858
  3. LC/MS/MS method for the quantitation of trans-2-hexenal-derived exocyclic 1,N(2)-propanodeoxyguanosine in DNA.
    Chem Res Toxicol. 2006 Apr;19(4):563-70 PMID: 16608168
  4. Detection of the carcinogen 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) in beer and wine.
    Carcinogenesis. 1993 May;14(5):899-901 PMID: 8504483
  5. Carcinogen-DNA adducts in human breast epithelial cells.
    Environ Mol Mutagen. 2002;39(2-3):184-92 PMID: 11921188
  6. UDP-glucuronosyltransferase activity in human liver and colon.
    Gastroenterology. 1999 Jan;116(1):149-60 PMID: 9869613
  7. Analysis and quantification of DNA adducts of 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline in liver of rats by liquid chromatography/electrospray tandem mass spectrometry.
    Chem Res Toxicol. 2002 Apr;15(4):551-61 PMID: 11952342
  8. DNA adduct measurements and tumor incidence during chronic carcinogen exposure in animal models: implications for DNA adduct-based human cancer risk assessment.
    Chem Res Toxicol. 1992 Nov-Dec;5(6):749-55 PMID: 1489923
  9. Reduction of signal suppression effects in ESI-MS using a nanosplitting device.
    Anal Chem. 2001 Dec 1;73(23):5635-44 PMID: 11774901
  10. Quantitation of pyridyloxobutyl DNA adducts of tobacco-specific nitrosamines in rat tissue DNA by high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry.
    Chem Res Toxicol. 2006 May;19(5):674-82 PMID: 16696570
  11. Standardization and validation of DNA adduct postlabelling methods: report of interlaboratory trials and production of recommended protocols.
    Mutagenesis. 1999 May;14(3):301-15 PMID: 10374998
  12. Determination of in vitro formed DNA adducts of 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine using capillary liquid chromatography/electrospray ionization/tandem mass spectrometry.
    Chem Res Toxicol. 1995 Dec;8(8):1005-13 PMID: 8605282
  13. Postlabeling methods for carcinogen-DNA adduct analysis.
    Environ Health Perspect. 1985 Oct;62:57-65 PMID: 3910421
  14. Nuclear distribution of aflatoxin B1 and its interaction with histones in rat liver in vivo.
    Cancer Res. 1980 Dec;40(12):4343-51 PMID: 7438068
  15. Mechanistic investigation of ionization suppression in electrospray ionization.
    J Am Soc Mass Spectrom. 2000 Nov;11(11):942-50 PMID: 11073257
  16. An unusual DNA adduct derived from the powerfully mutagenic environmental contaminant 3-nitrobenzanthrone.
    Chem Res Toxicol. 1998 Dec;11(12):1460-7 PMID: 9860488
  17. Identification of N-(Deoxyguanosin-8-yl)-2-amino-1-methyl-6-phenylimidazo [4,5-b]pyridine as the major adduct formed by the food-borne carcinogen, 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine, with DNA.
    Chem Res Toxicol. 1992 Sep-Oct;5(5):691-7 PMID: 1446011
  18. Immunological methods for detection of carcinogen-DNA damage in humans.
    Cancer Epidemiol Biomarkers Prev. 1999 Sep;8(9):733-9 PMID: 10498391
  19. DNA adducts: mass spectrometry methods and future prospects.
    Toxicol Appl Pharmacol. 2005 Sep 1;207(2 Suppl):293-301 PMID: 15990134
  20. Covalent DNA damage in tissues of cigarette smokers as determined by 32P-postlabeling assay.
    J Natl Cancer Inst. 1989 Mar 1;81(5):341-7 PMID: 2915370
  21. Comparison of the Paul ion trap to the linear ion trap for use in global proteomics.
    Proteomics. 2006 Mar;6(6):1735-40 PMID: 16475232
  22. Detection and quantification of N-(deoxyguanosin-8-yl)-4-aminobiphenyl adducts in human pancreas tissue using capillary liquid chromatography-microelectrospray mass spectrometry.
    Chem Res Toxicol. 2005 Apr;18(4):692-9 PMID: 15833029
  23. Development of a method for quantification of acrolein-deoxyguanosine adducts in DNA using isotope dilution-capillary LC/MS/MS and its application to human brain tissue.
    Anal Chem. 2005 Sep 15;77(18):5982-9 PMID: 16159131
  24. Accelerator mass spectrometry in biomedical dosimetry: relationship between low-level exposure and covalent binding of heterocyclic amine carcinogens to DNA.
    Proc Natl Acad Sci U S A. 1990 Jul;87(14):5288-92 PMID: 2371271
  25. 32P-postlabeling analysis of DNA adducts persisting for up to 42 weeks in the skin, epidermis and dermis of mice treated topically with 7,12-dimethylbenz[a]anthracene.
    Carcinogenesis. 1985 Aug;6(8):1117-26 PMID: 3926335
  26. Formation of DNA adducts of 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) in male Fischer-344 rats.
    Cancer Lett. 1992 Dec 24;67(2-3):117-24 PMID: 1483260
  27. 32P-post-labelling analysis of DNA adducts formed by food-derived heterocyclic amines: evidence for incomplete hydrolysis and a procedure for adduct pattern simplification.
    Carcinogenesis. 1994 May;15(5):877-82 PMID: 8200090
  28. Human cytochrome P-450PA (P-450IA2), the phenacetin O-deethylase, is primarily responsible for the hepatic 3-demethylation of caffeine and N-oxidation of carcinogenic arylamines.
    Proc Natl Acad Sci U S A. 1989 Oct;86(20):7696-700 PMID: 2813353
  29. Some current perspectives on chemical carcinogenesis in humans and experimental animals: Presidential Address.
    Cancer Res. 1978 Jun;38(6):1479-96 PMID: 348302
  30. Quantification of benzo[a]pyrene diol epoxide DNA-adducts by stable isotope dilution liquid chromatography/tandem mass spectrometry.
    Rapid Commun Mass Spectrom. 2006;20(8):1369-80 PMID: 16557497
  31. A two-dimensional quadrupole ion trap mass spectrometer.
    J Am Soc Mass Spectrom. 2002 Jun;13(6):659-69 PMID: 12056566
  32. Detection of 2-amino-1-methyl-6-phenylimidazo[4,5-b]-pyridine-DNA adducts in normal breast tissues and risk of breast cancer.
    Cancer Epidemiol Biomarkers Prev. 2003 Sep;12(9):830-7 PMID: 14504191
  33. Quantification of the heterocyclic aromatic amine DNA adduct N-(deoxyguanosin-8-yl)-2-amino-3-methylimidazo[4,5-f]quinoline in livers of rats using capillary liquid chromatography/microelectrospray mass spectrometry: a dose-response study.
    Anal Chem. 2001 Jul 1;73(13):2819-27 PMID: 11467522
  34. Quantitative analysis of 4-aminobiphenyl-C8-deoxyguanosyl DNA adducts produced in vitro and in vivo using HPLC-ES-MS.
    Carcinogenesis. 1999 Jun;20(6):1055-61 PMID: 10357788
  35. Biochemistry, biology, and carcinogenicity of tobacco-specific N-nitrosamines.
    Chem Res Toxicol. 1998 Jun;11(6):559-603 PMID: 9625726
  36. Fluorescence line-narrowing spectroscopy in the study of chemical carcinogenesis.
    Anal Chem. 1989 Sep 15;61(18):1023A-1024A, 1026A-1029A, 1031A-103 PMID: 2802152
  37. Determination of in vitro- and in vivo-formed DNA adducts of 2-amino-3-methylimidazo[4,5-f]quinoline by capillary liquid chromatography/microelectrospray mass spectrometry.
    Chem Res Toxicol. 1999 Oct;12(10):1019-27 PMID: 10525280
  38. Heterocyclic aromatic amines induce DNA strand breaks and cell transformation.
    Carcinogenesis. 1999 Apr;20(4):545-51 PMID: 10223180
  39. Extraction and purification of depurinated benzo[a]pyrene-adducted DNA bases from human urine by immunoaffinity chromatography coupled with HPLC and analysis by LC/quadrupole ion-trap MS.
    Chem Res Toxicol. 2003 Apr;16(4):479-86 PMID: 12703964
  40. Differentiation of isomeric C8-substituted alkylaniline adducts of guanine by electrospray ionization and tandem quadrupole ion trap mass spectrometry.
    J Am Soc Mass Spectrom. 2003 Dec;14(12):1488-92 PMID: 14652195
  41. Synthesis and spectroscopic characterization of site-specific 2-amino-1-methyl-6-phenylimidazo.
    Nucleic Acids Res. 2001 May 1;29(9):1951-9 PMID: 11328879
  42. Strategies for the assessment of matrix effect in quantitative bioanalytical methods based on HPLC-MS/MS.
    Anal Chem. 2003 Jul 1;75(13):3019-30 PMID: 12964746
  43. High-performance liquid chromatography electrospray ionization tandem mass spectrometry for the detection and quantitation of benzo[a]pyrene-DNA adducts.
    Chem Res Toxicol. 2005 Aug;18(8):1306-15 PMID: 16097804
  44. Effects of human and rat glutathione S-transferases on the covalent DNA binding of the N-acetoxy derivatives of heterocyclic amine carcinogens in vitro: a possible mechanism of organ specificity in their carcinogenesis.
    Cancer Res. 1994 Sep 15;54(18):4920-6 PMID: 8069858
  45. Hydroxyl free radical adduct of deoxyguanosine: sensitive detection and mechanisms of formation.
    Free Radic Res Commun. 1986;1(3):163-72 PMID: 2577733
  46. Formation and persistence of DNA adducts of 2-amino-3-methylimidazo[4,5-f]quinoline in the rat and nonhuman primates.
    Mutat Res. 1997 May 12;376(1-2):235-41 PMID: 9202760
  47. Methods development toward the measurement of polyaromatic hydrocarbon-DNA adducts by mass spectrometry.
    Res Rep Health Eff Inst. 1993 Oct;(61):1-25; discussion 27-36 PMID: 8267901
  48. Quantification of etheno-DNA adducts using liquid chromatography, on-line sample processing, and electrospray tandem mass spectrometry.
    Chem Res Toxicol. 2000 Dec;13(12):1259-64 PMID: 11123967
  49. Detection and quantification of endogenous cyclic DNA adducts derived from trans-4-hydroxy-2-nonenal in human brain tissue by isotope dilution capillary liquid chromatography nanoelectrospray tandem mass spectrometry.
    Chem Res Toxicol. 2006 May;19(5):710-8 PMID: 16696574
  50. Detection of guanine-C8-2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine adduct as a single spot on thin-layer chromatography by modification of the 32P-postlabeling method.
    Jpn J Cancer Res. 1994 Feb;85(2):113-7 PMID: 8144391
  51. Analysis of DNA adducts of 2-amino-1-methyl-6-phenylimidazo[4,5- b]pyridine in rat and human tissues by alkaline hydrolysis and gas chromatography/electron capture mass spectrometry: validation by comparison with 32P-postlabeling.
    Chem Res Toxicol. 1994 Nov-Dec;7(6):733-9 PMID: 7696526
  52. Liquid chromatography-electrospray ionization-mass spectrometry: the future of DNA adduct detection.
    Carcinogenesis. 2006 Feb;27(2):178-96 PMID: 16272169
  53. Genetically modified Chinese hamster ovary cells for investigating sulfotransferase-mediated cytotoxicity and mutation by 2-amino-1-methyl-6- phenylimidazo[4,5-b]pyridine.
    Environ Mol Mutagen. 2000;35(1):57-65 PMID: 10692228
  54. Formation and analysis of heterocyclic aromatic amine-DNA adducts in vitro and in vivo.
    J Chromatogr B Analyt Technol Biomed Life Sci. 2004 Mar 25;802(1):155-66 PMID: 15036007
  55. Characterization of DNA adducts formed in vitro by reaction of N-hydroxy-2-amino-3-methylimidazo[4,5-f]quinoline and N-hydroxy-2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline at the C-8 and N2 atoms of guanine.
    Chem Res Toxicol. 1992 Jul-Aug;5(4):479-90 PMID: 1391614
  56. High concentrations of the carcinogen 2-amino-1-methyl-6-phenylimidazo- [4,5-b]pyridine (PhIP) occur in chicken but are dependent on the cooking method.
    Cancer Res. 1995 Oct 15;55(20):4516-9 PMID: 7553619
  57. Heterogeneous DNA adduct formation in vitro by the acetylated food mutagen 2-(acetoxyamino)-1-methyl-6-phenylimidazo[4,5-b]pyridine: a fluorescence spectroscopic study.
    Chem Res Toxicol. 1995 Jul-Aug;8(5):659-70 PMID: 7548748
  58. Heterocyclic amines: Mutagens/carcinogens produced during cooking of meat and fish.
    Cancer Sci. 2004 Apr;95(4):290-9 PMID: 15072585
  59. Reaction of the N2-acetoxy derivative of 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) with 2'-deoxyguanosine and DNA. Synthesis and identification of N2-(2'-deoxyguanosin-8-yl)-PhIP.
    Carcinogenesis. 1992 Apr;13(4):629-35 PMID: 1576716
  60. Identification of DNA adducts using HPLC/MS/MS following in vitro and in vivo experiments with arylamines and nitroarenes.
    Chem Res Toxicol. 2003 Oct;16(10):1251-63 PMID: 14565767
  61. Electrospray ionization-tandem mass spectrometry and 32P-postlabeling analyses of tamoxifen-DNA adducts in humans.
    J Natl Cancer Inst. 2004 Jul 21;96(14):1099-104 PMID: 15265972
  62. Mass spectrometric detection and measurement of N2-(2'-deoxyguanosin-8-yl)PhIP adducts in DNA.
    J Chromatogr B Biomed Sci Appl. 2000 Jul 7;744(1):55-64 PMID: 10985566
  63. Detection of a carcinogen, 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), in cigarette smoke condensate.
    Carcinogenesis. 1991 Oct;12(10):1945-7 PMID: 1934275
  64. DNA adducts of heterocyclic amine food mutagens: implications for mutagenesis and carcinogenesis.
    Carcinogenesis. 1999 Mar;20(3):353-68 PMID: 10190547
  65. Genetic polymorphisms and modulation of 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP)-DNA adducts in human lymphocytes.
    Int J Cancer. 2003 Dec 20;107(6):878-84 PMID: 14601045
  66. DNA adduct formation from acrylamide via conversion to glycidamide in adult and neonatal mice.
    Chem Res Toxicol. 2003 Oct;16(10):1328-37 PMID: 14565774
  67. Applications of mass spectrometry for quantitation of DNA adducts.
    J Chromatogr B Analyt Technol Biomed Life Sci. 2002 Oct 5;778(1-2):323-43 PMID: 12376138
  68. Metabolic activation of carcinogenic heterocyclic aromatic amines by human liver and colon.
    Carcinogenesis. 1991 Oct;12(10):1839-45 PMID: 1934265
Article Info
Journal
Chemical research in toxicology
Abbr.
Chem Res Toxicol
ISSN
0893-228X
DOI
10.1021/tx0601713
Published
2007-02-00
Pages
263-76
Language
English
Region
United States
NLM ID
8807448
PMCID
PMC2593646
Subset
IM
Grants
NIEHS NIH HHS · R21 ES014438 · United States
NIEHS NIH HHS · R21 ES014438-01A1 · United States
External Links
PubMed source DOI: 10.1021/tx0601713 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2