Analytical and Bioanalytical Chemistry

, Volume 402, Issue 9, pp 2749–2763 | Cite as

Internal correction of spectral interferences and mass bias for selenium metabolism studies using enriched stable isotopes in combination with multiple linear regression

  • Kristoffer Lunøe
  • Justo Giner Martínez-Sierra
  • Bente Gammelgaard
  • J. Ignacio García AlonsoEmail author
Original Paper


The analytical methodology for the in vivo study of selenium metabolism using two enriched selenium isotopes has been modified, allowing for the internal correction of spectral interferences and mass bias both for total selenium and speciation analysis. The method is based on the combination of an already described dual-isotope procedure with a new data treatment strategy based on multiple linear regression. A metabolic enriched isotope (77Se) is given orally to the test subject and a second isotope (74Se) is employed for quantification. In our approach, all possible polyatomic interferences occurring in the measurement of the isotope composition of selenium by collision cell quadrupole ICP-MS are taken into account and their relative contribution calculated by multiple linear regression after minimisation of the residuals. As a result, all spectral interferences and mass bias are corrected internally allowing the fast and independent quantification of natural abundance selenium (natSe) and enriched 77Se. In this sense, the calculation of the tracer/tracee ratio in each sample is straightforward. The method has been applied to study the time-related tissue incorporation of 77Se in male Wistar rats while maintaining the natSe steady-state conditions. Additionally, metabolically relevant information such as selenoprotein synthesis and selenium elimination in urine could be studied using the proposed methodology. In this case, serum proteins were separated by affinity chromatography while reverse phase was employed for urine metabolites. In both cases, 74Se was used as a post-column isotope dilution spike. The application of multiple linear regression to the whole chromatogram allowed us to calculate the contribution of bromine hydride, selenium hydride, argon polyatomics and mass bias on the observed selenium isotope patterns. By minimising the square sum of residuals for the whole chromatogram, internal correction of spectral interferences and mass bias could be accomplished. As a result, the tracer/tracee ratio could be calculated for each selenium-containing species and a time relationship for synthesis and degradation established. Both selenite and selenized yeast labelled with 77Se were employed for comparative purposes.


Mass spectrometry ICP-MS Speciation Biological samples 



We would like to thank PharmaNord for supplying of the 77Se enriched yeast, Marisa Fernández Sánchez for the supply of the 74Se spike and Teresa Fernández and Agustín Brea from the Biotery of the University of Oviedo for their help regarding animal experiments. Funding from the Spanish Ministry of Science and Innovation through project number CTQ2009-12814 and the Education and Science Council of the Principado de Asturias (grant BP07-059) are gratefully acknowledged.


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Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • Kristoffer Lunøe
    • 1
  • Justo Giner Martínez-Sierra
    • 2
  • Bente Gammelgaard
    • 1
  • J. Ignacio García Alonso
    • 2
    Email author
  1. 1.Department of Pharmaceutics and Analytical Chemistry, Faculty of Pharmaceutical SciencesUniversity of CopenhagenCopenhagenDenmark
  2. 2.Department of Physical and Analytical ChemistryUniversity of OviedoOviedoSpain

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