Advertisement

Analytical and Bioanalytical Chemistry

, Volume 406, Issue 5, pp 1519–1530 | Cite as

Conditions for sample preparation and quantitative HPLC/MS-MS analysis of bulky adducts to serum albumin with diolepoxides of polycyclic aromatic hydrocarbons as models

  • Emelie Westberg
  • Ulla Hedebrant
  • Johanna Haglund
  • Tomas Alsberg
  • Johan Eriksson
  • Albrecht Seidel
  • Margareta Törnqvist
Research Paper

Abstract

Stable adducts to serum albumin (SA) from electrophilic and genotoxic compounds/metabolites can be used as biomarkers for quantification of the corresponding in vivo dose. In the present study, conditions for specific analysis of stable adducts to SA formed from carcinogenic polycyclic aromatic hydrocarbons (PAH) were evaluated in order to achieve a sensitive and reproducible quantitative method. Bulky adducts from diolepoxides (DE) of PAH, primarily DE of benzo[a]pyrene (BPDE) and also DE of dibenzo[a,l]pyrene (DBPDE) and dibenzo[a,h]anthracene (DBADE), were used as model compounds. The alkylated peptides obtained after enzymatic hydrolysis of human SA modified with the different PAHDE were principally PAHDE-His-Pro, PAHDE-His-Pro-Tyr and PAHDE-Lys. Alkaline hydrolysis under optimised conditions gave the BPDE-His as the single analyte of alkylated His, but also indicated degradation of this adduct. It was not possible to obtain the BPDE-His as one analyte from BPDE-alkylated SA through modifications of the enzymatic hydrolysis. The BPDE-His adduct was shown to be stable during the weak acidic conditions used in the isolation of SA. Enrichment by HPLC or SPE, but not butanol extraction, gave good recovery, using Protein LoBind tubes. A simple internal standard (IS) approach using SA modified with other PAHDE as IS was shown to be applicable. A robust analytical procedure based on digestion with pronase, enrichment by HPLC or SPE, and analysis with HPLC/MS-MS electrospray ionisation was achieved. A good reproducibility (coefficient of variation (CV) 11 %) was obtained, and the achieved limit of detection for the studied PAHDE, using standard instrumentation, was approximately 1 fmol adduct/mg SA analysing extract from 5 mg SA.

Figure

An outline of the method for analysis of bulky SA-adducts. All steps/conditions were evaluated.

Keywords

Bulky serum albumin adducts Polycyclic aromatic hydrocarbons Extraction (SPE | HPLC | butanol) Diol epoxides Mass Spectrometry Hydrolysis (pronase enzymatic | alkaline) 

Notes

Acknowledgments

This work has been carried out with economical support from the European Commission; contract no. QLK4-CT-2002-02402 (AMBIPAH), the Swedish Research Council FORMAS and the Swedish Cancer and Allergy Foundation. We would like to thank Dr. Hans Helleberg for valuable scientific advice at the initiation of the study.

Supplementary material

216_2013_7540_MOESM1_ESM.pdf (194 kb)
ESM 1 (PDF 193 KB)

References

  1. 1.
    Rubino FM, Pitton M, Di Fabio D, Colombi A (2009) Mass Spectrom Rev 28:725–784CrossRefGoogle Scholar
  2. 2.
    Tornqvist M, Fred C, Haglund J, Helleberg H, Paulsson B, Rydberg P (2002) J Chromatogr B Anal Technol Biomed Life Sci 778:279–308CrossRefGoogle Scholar
  3. 3.
    Boogaard PJ (2002) J Chromatogr B Anal Technol Biomed Life Sci 778:309–322CrossRefGoogle Scholar
  4. 4.
    Rappaport SM, Yeowell-O'Connell K, Smith MT, Dosemeci M, Hayes RB, Zhang L, Li G, Yin S, Rothman N (2002) J Chromatogr B Anal Technol Biomed Life Sci 778:367–374CrossRefGoogle Scholar
  5. 5.
    Sabbioni G, Jones CR (2002) Biomarkers 7:347–421CrossRefGoogle Scholar
  6. 6.
    Chapot B, Wild CP (1991) Tech Diagn Pathol 2:135–155Google Scholar
  7. 7.
    Gong YY, Wilson S, Mwatha JK, Routledge MN, Castelino JM, Zhao B, Kimani G, Kariuki HC, Vennervald BJ, Dunne DW, Wild CP (2012) Environ Health Perspect 120:893–896CrossRefGoogle Scholar
  8. 8.
    International Agency for Research on Cancer (IARC) (2010) Some non-heterocyclic polycyclic aromatic hydrocarbons and some related exposures, Monographs on the Evaluation of Carcinogenic Risks to Humans, vol 92. IARC, LyonGoogle Scholar
  9. 9.
    National Food Agency (2010) Market Basket 2010 - chemical analysis, exposure estimation and health-related assessment of nutrients and toxic compounds in Swedish food baskets, report no 7/2012. National Food Agency, SwedenGoogle Scholar
  10. 10.
    Conney AH, Chang RL, Jerina DM, Wei S-JC (1994) Drug Metab Rev 26:125–163CrossRefGoogle Scholar
  11. 11.
    Hall M, Forrester LM, Parker DK, Grover PL, Wolf CR (1989) Carcinogenesis 10:1815–1821CrossRefGoogle Scholar
  12. 12.
    Slaga TJ, Bracken WJ, Gleason G, Levin W, Yagi H, Jerina DM, Conney AH (1979) Cancer Res 39:67–71Google Scholar
  13. 13.
    Day BW, Skipper PL, Zaia J, Singh K, Tannenbaum SR (1994) Chem Res Toxicol 7:829–835CrossRefGoogle Scholar
  14. 14.
    Day BW, Skipper PL, Rich RH, Naylor S, Tannenbaum SR (1991) Chem Res Toxicol 4:359–363CrossRefGoogle Scholar
  15. 15.
    Skipper PL, Naylor S, Gan LS, Day BW, Pastorelli R, Tannenbaum SR (1989) Chem Res Toxicol 2:280–281CrossRefGoogle Scholar
  16. 16.
    Naylor S, Gan LS, Day BW, Pastorelli R, Skipper PL, Tannenbaum SR (1990) Chem Res Toxicol 3:111–117CrossRefGoogle Scholar
  17. 17.
    Helleberg H, Tornqvist M (2000) Rapid Commun Mass Spectrom 14:1644–1653CrossRefGoogle Scholar
  18. 18.
    Day BW, Skipper PL, Zaia J, Tannenbaum SR (1991) J Am Chem Soc 113:8505–8509CrossRefGoogle Scholar
  19. 19.
    Kaefferlein HU, Marczynski B, Mensing T, Bruening T (2010) Crit Rev Toxicol 40:126–150CrossRefGoogle Scholar
  20. 20.
    Ozbal CC, Skipper PL, Yu MC, London SJ, Dasari RR, Tannenbaum SR (2000) Cancer Epidemiol Biomarkers Prev 9:733–734Google Scholar
  21. 21.
    Luch A, Glatt H, Platt K, Oesch F, Seidel A (1994) Carcinogenesis 15:2507–2516CrossRefGoogle Scholar
  22. 22.
    Lee HM, Harvey RG (1980) J Org Chem 45:588–592CrossRefGoogle Scholar
  23. 23.
    Weinstein IB, Jeffrey AM, Jennette KW, Blobstein SH, Harvey RG, Harris C, Autrup H, Kasai H, Nakanishi K (1976) Science 193:592–595CrossRefGoogle Scholar
  24. 24.
    Luch A, Platt KL, Seidel A (1998) Carcinogenesis 19:639–648CrossRefGoogle Scholar
  25. 25.
    Zaia J, Biemann K (1994) J Am Chem Soc 116:7407–7408CrossRefGoogle Scholar
  26. 26.
    Brunmark P, Harriman S, Skipper PL, Wishnok JS, Amin S, Tannenbaum SR (1997) Chem Res Toxicol 10:880–886CrossRefGoogle Scholar
  27. 27.
    Chung MK, Regazzoni LG, McClean M, Herrick R, Rappaport SM (2013) Anal Biochem 435(2):140–149Google Scholar
  28. 28.
    Gupta RC (1985) Cancer Res 45:5656–5662Google Scholar
  29. 29.
    Vikstrom AC, Abramsson-Zetterberg L, Naruszewicz M, Athanassiadis I, Granath FN, Tornqvist M (2011) Toxicol Sci 119:41–49Google Scholar
  30. 30.
    von Stedingk H, Vikstrom AC, Rydberg P, Pedersen M, Nielsen JKS, Segerback D, Knudsen LE, Tornqvist M (2011) Chem Res Toxicol 24:1957–1965CrossRefGoogle Scholar
  31. 31.
    Damsten MC, Commandeur JNM, Fidder A, Hulst AG, Touw D, Noort D, Vermeulen NPE (2007) Drug Metab Dispos 35:1408–1417CrossRefGoogle Scholar
  32. 32.
    Fustinoni S, Colosio C, Colombi A, Lastrucci L, Yeowell-O'Connell K, Rappaport SM (1998) Int Arch Occup Environ Health 71:35–41CrossRefGoogle Scholar
  33. 33.
    Rappaport SM, Ting D, Jin Z, Yeowell-O'Connell K, Waidyanatha S, McDonald T (1993) Chem Res Toxicol 6:238–244CrossRefGoogle Scholar
  34. 34.
    Noort D, Hulst AG, De Jong LPA, Benschop HP (1999) Chem Res Toxicol 12:715–721CrossRefGoogle Scholar
  35. 35.
    Waidyanatha S, Zheng Y, Serdar B, Rappaport SM (2004) Cancer Epidemiol Biomarkers Prev 13:117–124CrossRefGoogle Scholar
  36. 36.
    Singh R, Gaskell M, Le Pla RC, Kaur B, Zim-Araghi A, Roach J, Koukouves G, Souliotis VL, Kyrtopoulos SA, Farmer PB (2006) Chem Res Toxicol 19:868–878CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2014

Authors and Affiliations

  • Emelie Westberg
    • 1
  • Ulla Hedebrant
    • 1
  • Johanna Haglund
    • 1
    • 4
  • Tomas Alsberg
    • 2
  • Johan Eriksson
    • 1
  • Albrecht Seidel
    • 3
  • Margareta Törnqvist
    • 1
  1. 1.Division of Environmental Chemistry, Department of Materials and Environmental ChemistryStockholm UniversityStockholmSweden
  2. 2.Department of Applied Environmental ScienceStockholm UniversityStockholmSweden
  3. 3.Biochemical Institute for Environmental CarcinogensProf. Dr. Gernot Grimmer-FoundationGrosshansdorfGermany
  4. 4.MetaSafe ABSödertäljeSweden

Personalised recommendations