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Simultaneous speciation analysis of glutathione peroxidase, selenoprotein P and selenoalbumin in human serum by tandem anion exchange-affinity HPLC and on-line isotope dilution ICP-quadrupole MS

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Abstract

A method based on anion exchange (AE) and affinity (AF)-HPLC (AE-AF-HPLC) hyphenated to inductively coupled plasma-(quadrupole) mass spectrometry (ICP-QMS) was developed for the speciation analysis of selenoprotein P (SelP), glutathione peroxidase (GPx) and selenoalbumin (SeAlb) in human serum. AE-HPLC is proposed here for the on-line alleviation of Cl and Br spectral interferences on 77Se (40Ar37Cl) and 82Se (81Br1H). Separation of GPx, SelP and SeAlb by AE-AF-HPLC was obtained within a total chromatographic runtime of <20 min. On-line (post-column) isotope dilution (ON-ID) and on-line external calibration (ON-EC)-ICP-QMS were used for the quantification of Se in GPx, SelP and SeAlb. ON-EC using a Se-L-cystine standard was shown to be a suitable approach for the routine simultaneous speciation analysis of serum GPx, SelP and SeAlb. The method validation was carried out by direct ICP-sector field MS determination of Se in GPx, SelP and SeAlb fractions collected after AE-AF-HPLC separation. In addition, the method accuracy for the determination of total protein-bound Se was assessed by analyzing a human serum reference material (BCR-637) certified for total Se content.

A methodology for the alleviation of Cl and Br interferences in the accurate simultaneous speciation analysis of glutathione peroxidase, selenoprotein P and selenoalbumin in human serum by affinity HPLC coupled on-line with ICP-quadrupole MS is proposed. This approach may be particularly useful for clinical laboratories that only have an ICP-quadrupole MS without a collision cell, or that lack an expensive ICP-SFMS (high-resolution) instrument 

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References

  1. Hatfield DL, Berry MJ, Gladyshev VN (eds)(2006) Selenium: its molecular biology and role in human health. Springer, New York

  2. Reilly C (2006) Selenium in food and health. Springer, New York

  3. Dumont E, Vanhaecke F, Cornelis R (2006) Anal Bioanal Chem 385:1304–1323

    Article  Google Scholar 

  4. Gromer S, Eubel JK, Lee BL, Jacob J (2005) Cell Mol Life Sci 62:2414–2437

    Article  CAS  Google Scholar 

  5. Gao Y, Wang Z (2000) Fresenius J Anal Chem 367:60–64

    Article  CAS  Google Scholar 

  6. Hinojosa Reyes L, Marchante-Gayon JM, Garcia Alonso JI, Sanz-Medel A (2003) J Anal Atom Spectrom 18:1210–1216

    Google Scholar 

  7. Jitaru P, Prete M, Cozzi G, Turetta C, Cairns W, Seraglia R, Traldi P, Cescon P, Barbante C (2008) J Anal Atom Spectrom 23:402–406

    Google Scholar 

  8. Shigeta K, Sato K, Furuta N (2007) J Anal Atom Spectrom 22:911–916

    Google Scholar 

  9. Miller ME, Cappon C (1984) J Clin Chem 30(5):781–783

    CAS  Google Scholar 

  10. Palacios O, Ruiz Encinar J, Schaumlöffel D, Lobinski R (2006) Anal Bioanal Chem 384:1276–1283

    Article  Google Scholar 

  11. Turner J, Hill SJ, Evans EH, Fairmana B, Wolff Briche CSJ (2000) J Anal Atom Spectrom 15:743–746

    Google Scholar 

  12. Rottman L, Heumann KG (1994) Fresenius J Anal Chem 350:221–227

    Article  Google Scholar 

  13. Heumann KG, Gallus SM, Radlinger G, Vogl J (1998) J Anal Atom Spectrom 13:1001–1008

    Google Scholar 

  14. Sariego Muniz C, Marchante Gayon JM, Garcia Alonso JI, Sanz-Medel A (2001) J Anal Atom Spectrom 16:587–592

    Google Scholar 

  15. Schaumlöffel D, Prange A, Marx G, Heumann KG, Bratter P (2002) Anal Bioanal Chem 372:155–163

    Article  Google Scholar 

  16. Goenaga Infante H, Van Campenhout K, Schaumlöffel D, Blust R, Adams FC (2003) Analyst 128:651–657

    Article  CAS  Google Scholar 

  17. Rodriguez-Gonzalez P, Marchante-Gayon JM, Garcia Alonso JI, Sanz-Medel A (2005) Spectrochim Acta Part B 60:151–207

    Google Scholar 

Download references

Acknowledgements

This study is a contribution to the Marie-Curie Intra-European Project (MEIF-CT-2006–024156/ELSA-BIM) funded by the European Commission. Dr. Pietro Traldi (ISTM-CNR, Padova, Italy) and Dr. Annunziata Lapolla (University of Padova, Department of Metabolism Diseases, Italy) are acknowledged for their useful scientific discussions. The authors thank Dr. Clara Turetta (IDPA-CNR) and Marco Roman (University of Venice Ca’ Foscari) for laboratory assistance. ELGA LabWater is acknowledged for providing the PURELAB Option-R and Ultra Analytic systems, which produced the ultrapure water used in these experiments. The Departments of Immunohaematology of Azienda ULSS 2 di Feltre (BL) and Servizio Immunoematologia e Trasfusionale ULSS 12 Veneziana Ospedale Umberto I, Mestre (VE), Italy are also acknowledged for providing the control serum.

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Authors and Affiliations

  1. Institute for the Dynamics of Environmental Processes (IDPA-CNR), Dorsoduro 2137, 30123, Venice, Italy

    Petru Jitaru, Andrea Gambaro, Paolo Cescon & Carlo Barbante

  2. Department of Inorganic and Analytical Chemistry, Faculty of Chemistry, University “Al. I. Cuza”, 11 Carol I Blvd., 700506, Iasi, Romania

    Petru Jitaru

  3. Department of Environmental Sciences, University of Venice Ca’ Foscari, Dorsoduro 2137, 30123, Venice, Italy

    Giulio Cozzi, Andrea Gambaro, Paolo Cescon & Carlo Barbante

Authors
  1. Petru Jitaru
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  2. Giulio Cozzi
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  3. Andrea Gambaro
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  4. Paolo Cescon
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  5. Carlo Barbante
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Correspondence to Carlo Barbante.

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Jitaru, P., Cozzi, G., Gambaro, A. et al. Simultaneous speciation analysis of glutathione peroxidase, selenoprotein P and selenoalbumin in human serum by tandem anion exchange-affinity HPLC and on-line isotope dilution ICP-quadrupole MS. Anal Bioanal Chem 391, 661–669 (2008). https://doi.org/10.1007/s00216-008-2043-7

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  • DOI: https://doi.org/10.1007/s00216-008-2043-7

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