Issue 9, 2016

Accurate quantification of selenoproteins in human plasma/serum by isotope dilution ICP-MS: focus on selenoprotein P

Abstract

A species-specific isotope dilution analysis (SS IDA) method was developed for the first time for the determination of selenoprotein P (SEPP1) in human plasma/serum at the protein level by double affinity HPLC-ICP-MS. In this regard, a standard and a spike of SEPP1 were produced by cell-free E. coli protein synthesis, where Se (ICP tag) was introduced in the form of selenomethionine (SeMet) allowing for the absolute SEPP1 quantification by ICP-MS. A complete characterization of the standard and the spike was carried out in terms of isotopic composition and Se mass fraction by collision-cell ICP-MS to ensure SI-traceability. Method development and validation were conducted using the reference materials BCR-637, extensively analysed for its Se species, and the SRM 1950, which provides reference values for Se species (including SEPP1). Stability of the isotope ratio R77/76 in the sample blends was tested for one month with negligible change. Relative expanded uncertainties of 5.7% and 7.7% were achieved for BCR-637 and SRM 1950 for mass fractions of 55.5 and 63.9 ng g−1 Se for SEPP1, respectively. The developed SEPP1 SS IDA methodology could be a valuable tool to establish references values for selenoproteins in clinical chemistry and showed the potential of cell-free protein synthesis for the preparation of future stable isotope-labeled intact selenoproteins.

Graphical abstract: Accurate quantification of selenoproteins in human plasma/serum by isotope dilution ICP-MS: focus on selenoprotein P

Supplementary files

Article information

Article type
Paper
Submitted
23 جمادى الثانية 1437
Accepted
21 شوال 1437
First published
21 شوال 1437

J. Anal. At. Spectrom., 2016,31, 1904-1912

Accurate quantification of selenoproteins in human plasma/serum by isotope dilution ICP-MS: focus on selenoprotein P

M. E. D. C. Busto, C. Oster, S. Cuello-Nuñez, C. L. Deitrich, A. Raab, A. Konopka, W. D. Lehmann, H. Goenaga-Infante and P. Fisicaro, J. Anal. At. Spectrom., 2016, 31, 1904 DOI: 10.1039/C6JA00122J

To request permission to reproduce material from this article, please go to the Copyright Clearance Center request page.

If you are an author contributing to an RSC publication, you do not need to request permission provided correct acknowledgement is given.

If you are the author of this article, you do not need to request permission to reproduce figures and diagrams provided correct acknowledgement is given. If you want to reproduce the whole article in a third-party publication (excluding your thesis/dissertation for which permission is not required) please go to the Copyright Clearance Center request page.

Read more about how to correctly acknowledge RSC content.

Social activity

Spotlight

Advertisements