Isolation of Selenoprotein-P and Determination of Se Concentration Incorporated in Proteins in Human and Mouse Plasma by Tandem Heparin Affinity and Size-exclusion Column HPLC-ICPMS

• SUZUKI Yoshinari

  Department of Applied Chemistry, Faculty of Science and Engineering, Chuo University

• SAKAI Tatsuya

  Department of Applied Chemistry, Faculty of Science and Engineering, Chuo University

• FURUTA Naoki

  Department of Applied Chemistry, Faculty of Science and Engineering, Chuo University

Abstract

Sel-P is considered to be the most important selenoprotein for evaluating the selenium (Se) status in the body. To isolate and determine Sel-P in plasma, we have developed an analytical method combining heparin affinity (AF) and size-exclusion column (SEC) high-performance liquid chromatography–inductively coupled plasma mass spectrometry (HPLC-ICPMS). We used this method to validate the adsorption efficiency of selenoproteins on a heparin AF column, and to then determine the Se concentrations incorporated in proteins in human and mouse plasma. The adsorption efficiency of Sel-P on a heparin column was more than 90% for both human and mouse plasma. Tandem AF and SEC separation proved to be useful for determining the Se concentrations incorporated in Sel-P in mouse plasma, but not in human plasma, because of nonspecific adsorption of plasma-extracellular glutathione peroxidase (eGPx) and albumin on the heparin AF column. Ultimately, we used the tandem AF and SEC separation method for mouse plasma and SEC separation alone for human plasma. The Se concentration incorporated in selenoproteins determined by our method showed good agreements with the total Se concentration determined following acid digestion.

Journal

• Analytical Sciences

  Analytical Sciences 28 (3), 221-221, 2012

  The Japan Society for Analytical Chemistry

Keywords

• Analytical Chemistry

Details

• CRID: 1390282679235905408
• NII Article ID: 130004442117
• NII Article ID: 10030446979
• NII Book ID: AA10500785
• DOI: 10.2116/analsci.28.221
• COI: 1:STN:280:DC%2BC38vpvVKhtQ%3D%3D
• ISSN: 13482246
• ISSN: 09106340
• NDL BIB ID: 023495722
• PubMed: 22451360
• Web Site: https://ndlsearch.ndl.go.jp/books/R000000004-I023495722
• Web Site: https://link.springer.com/content/pdf/10.2116/analsci.28.221.pdf
• Web Site: https://link.springer.com/article/10.2116/analsci.28.221/fulltext.html
• Web Site: https://www.jstage.jst.go.jp/article/analsci/28/3/28_3_221/_pdf
• Web Site: https://search.jamas.or.jp/link/ui/2012272982
• Text Lang: en
• Data Source:
  • JaLC
  • NDL
  • Crossref
  • PubMed
  • CiNii Articles
• Abstract License Flag: Disallowed