SpringerLink
Log in

Trace determination of peptides in water samples using packed capillary liquid chromatography with UV and MS detection and characterization of peptide oxidation products by MS

  • Original Paper
  • Published:
Analytical and Bioanalytical Chemistry Aims and scope Submit manuscript

Abstract

A capillary liquid chromatographic column switching method has been developed for fast and sensitive determination of peptides in water samples. Sample volumes of 1 mL were loaded onto a (320 μm I.D. ×30 mm) 10 μm Kromasil C18 pre-column, providing on-line analyte enrichment, prior to back-flushed elution onto a (320 μm I.D. ×150 mm) 3.5 μm Kromasil C18 analytical column. Loading flow rates of 250 μL/min and a mobile phase composition of acetonitrile/water/trifluoroacetic acid (22/77.9/0.1, v/v) provided a total analysis time of less than 25 minutes for the test peptides angiotensin II, bombesin, bradykinin, corazonin, neurotensin and substance P, using temperature programmed elution. In addition, solvent gradient elution and combined solvent gradient elution and temperature programming were explored. Using on-capillary UV detection at 210 nm resulted in a concentration limit of detection (cLOD) of about 1 ng/mL. The method was validated over the concentration range 1–100 ng/mL, yielding a coefficient of correlation of 0.997 or better. The within-assay (n=6) and between-assay (n=6) precisions of peak areas were on average 6% RSD and 5% RSD, respectively.

When the method was applied to spiked chlorinated tap water samples, it was found that peptides containing methionine, tryptophan and cystine were oxidized. Identification of the oxidation products of the peptides in hypochlorite-treated water was done with positive electrospray ionization time-of-flight mass spectrometric detection.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. Hancock JR, D’Agostino PA (2002) Anal Chim Acta 457:71

    Article  CAS  Google Scholar 

  2. Kientz CE (1998) J Chromatogr A 814:1

    Google Scholar 

  3. D’Agostino PA, Hancock JR, Provost LR (1997) J Chromatogr A 767:77

    Google Scholar 

  4. Kientz CE, Hulst AG, Wils ERJ (1997) J Chromatogr A 757:51

    Article  CAS  PubMed  Google Scholar 

  5. Molander P, Haugland K, Lundanes E, Thorud S, Thomassen Y, Greibrokk T (2000) J Chromatogr A 892:67

    Google Scholar 

  6. Holm A, Molander P, Lundanes E, Greibrokk T (2003) J Chromatogr A 983:43

    Article  CAS  PubMed  Google Scholar 

  7. Rezai MA, Famiglini G, Cappiello A (1996) J Chromatogr A 742:69

    Article  CAS  Google Scholar 

  8. Molander P, Haugland K, Hegna DR, Ommundsen E, Lundanes E, Greibrokk T (1999) J Chromatogr A 864:103

    PubMed  Google Scholar 

  9. van der Heeft E, Ten Hove GJ, Herberts CA, Meiring HD, Van Els CACM, De Jong APJM (1998) Anal Chem 70:3742

    Article  PubMed  Google Scholar 

  10. Pascual JA, Ten Hove GJ, De Jong APJM (1996) J Microcolumn Sep 8:383

    Article  CAS  Google Scholar 

  11. Oehrle SA, Westrick J (2002) LC GC Europe 15:728

    CAS  Google Scholar 

  12. Nawrocki J (1997) J Chromatogr A 779:29

    Google Scholar 

  13. Cai B, Li J (1999) Anal Chim Acta 399:249

    Article  CAS  Google Scholar 

  14. Pearson JD, McCroskey MC (1996) J Chromatogr A 746:277

    Google Scholar 

  15. Garcia MC, Hogenboom AC, Zappey H, Irth H (2002) J Chromatogr A 957:187

    Google Scholar 

  16. Mehlis B, Kertscher U (1997) Anal Chim Acta 352:71

    Article  CAS  Google Scholar 

  17. Hühmer AFR, Aced GI, Perkins MD, Güersoy RN, Jois DSS, Larive C, Siahan TJ, Schöneich C (1997) Anal Chem 69:29R

    Article  PubMed  Google Scholar 

  18. Chen H, Horvath C (1995) J Chromatogr A 705:3

    Google Scholar 

  19. Oosterkamp AJ, Carrascal M, Closa D, Escolar G, Gelpi E, Abian J (2001) J Microcolumn Sep 13:265

    Article  CAS  Google Scholar 

  20. Nightingale ZD, Lancha AHJ, Handelman SK, Dolnikowski GG, Busse SC, Dratz EA, Blumberg JB, Handelman GJ (2000) Free Radical Bio Med 29:425

    Article  CAS  Google Scholar 

  21. Kotiaho T, Eberlin MN, Vainiotalo P, Kostiainen R (2000) J Am Soc Mass Spectr 11:526

    Article  CAS  Google Scholar 

  22. Simat T, Meyer K, Steinhart H (1994) J Chromatogr A 661:93

    Article  CAS  Google Scholar 

  23. Nagalla SR, Barry BJ, Falick AM, Gibson BW, Taylor JE, Dong JZ, Spindel ER (1996) J Biol Chem 271:7731

    Article  CAS  PubMed  Google Scholar 

  24. Li S, Schöneich C, Wilson GS, Borchardt RT (1993) Pharmacol Res 10:1572

    Article  CAS  Google Scholar 

  25. Linde S, Nielsen JH, Hansen B, Welinder BS (1990) J Chromatogr 530:29

    Google Scholar 

  26. Dillen L, Zhang XY, Claeys M, Lianf F, De Potter WP, Van Dongen W, Esmans EL (1995) J Mass Spectrom 30:1599

    CAS  Google Scholar 

  27. Mudd JB, Leavitt R, Ongun A, McManus TT (1969) Atmos Environ 3:669

    Article  CAS  PubMed  Google Scholar 

  28. Morand K, Talbo G, Mann M (1993) Rapid Commun Mass Sp 7:738

    CAS  Google Scholar 

  29. Van de Weert M, Lagerwerf FM, Haverkamp J, Heerma W (1998) J Mass Spectrom 33:884

    Article  Google Scholar 

  30. Maleknia SD, Chance MR, Downard KM (1999) Rapid Commun Mass Sp 13:2352

    Article  CAS  Google Scholar 

  31. Hawkins CL, Davies MJ (1998) Biochem J 332:617

    CAS  PubMed  Google Scholar 

  32. Peskin AV, Winterbourn CC (2001) Free Radical Bio Med 30:572

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This research has been supported in part by the European Union programme “Energy, Environment and Sustainable Development” under Marie Curie Fellowship contract No. EVK1-CT-2000-57121, and the European Union programme “Improving Human Research Potential and the Socio-Economic Knowledge Base” under contract No. HPRN-CT-2001-00180.

Author information

Authors and Affiliations

  1. Department of Chemistry, University of Oslo, P.O. Box 1033, 0315, Blindern, Norway

    Thomas Bjellaas, Anders Holm, Paal Molander, Tyge Greibrokk & Elsa Lundanes

  2. National Institute of Occupational Health, P.O. Box 8149 Dep., 0033, Oslo, Norway

    Paal Molander

  3. Norwegian Defence Research Establishment, P.O. Box 25, 2027, Kjeller, Norway

    John Aasulf Tørnes

Authors
  1. Thomas Bjellaas
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Anders Holm
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Paal Molander
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. John Aasulf Tørnes
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Tyge Greibrokk
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Elsa Lundanes
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Elsa Lundanes.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Bjellaas, T., Holm, A., Molander, P. et al. Trace determination of peptides in water samples using packed capillary liquid chromatography with UV and MS detection and characterization of peptide oxidation products by MS. Anal Bioanal Chem 378, 1021–1030 (2004). https://doi.org/10.1007/s00216-003-2405-0

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00216-003-2405-0

Keywords

Search

Navigation