Analytical and Bioanalytical Chemistry

, Volume 399, Issue 3, pp 1141–1149 | Cite as

Simultaneous analysis of cardiac glycosides in blood and urine by thermoresponsive LC-MS-MS

  • Sanae Kanno
  • Kanako Watanabe
  • Itaru Yamagishi
  • Seishiro Hirano
  • Kayoko Minakata
  • Kunio Gonmori
  • Osamu Suzuki
Original Paper
First Online:
Received:
Revised:
Accepted:

Abstract

A new thermoresponsive polymer separation column was applied to simultaneous analysis of four cardiac glycosides (CGs) being widely used for the treatment of arrhythmias and heart failure in human blood and urine. This column is composed of an N-isopropylacrylamide polymer, the surface of which undergoes a reversible alteration from hydrophilic to hydrophobic by changing temperature. The chromatographic separation and retention times can be easily be controlled by adjusting the column temperature. As the column temperature was changed from 50 to 10 °C over 8 min, five CGs, including deslanoside, digoxin, methyldigoxin, digitoxin, and digitoxigenin (internal standard) were better resolved. Using these LC conditions, we analyzed four CGs in human whole blood and urine simultaneously by liquid chromatography-tandem mass spectrometry (LC-MS-MS). Validation data as functions of recovery rates, linearity, accuracy, and precision were carefully tested; all were generally satisfactory. The detection limits for the four CGs in both matrices were 0.2–0.3 ng/mL. The method was applied to analysis of methyldigoxin and its main metabolite digoxin in whole blood and urine samples obtained from a deceased person in actual autopsy case. To our knowledge, this is the first report describing an LC-MS-MS method using a thermoresponsive column for analysis of drug(s). The inclusion of the thermoresponsive column in an LC-MS-MS technique seems to extend the possibility for simultaneous analysis of compounds of different properties, such as hydrophobic precursors and their hydrophilic metabolites in biological samples within limited analysis times.

Keywords

Poly(N-isopropylacrylamide) Cardiac glycosides Thermoresponsive LC-MS-MS Digoxin Methyldigoxin 
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Notes

Acknowledgments

The present study was supported by the Ministry of Environment, Japan (The Development of New Environmental Technology—NANO-01). We would like to thank Ms. Masako Suzuki for technical assistance in MS-MS.

Supplementary material

216_2010_4405_MOESM1_ESM.pdf (68 kb)
ESM 1 (PDF 67 kb)

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Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Sanae Kanno
    • 1
    • 3
  • Kanako Watanabe
    • 1
  • Itaru Yamagishi
    • 1
  • Seishiro Hirano
    • 2
  • Kayoko Minakata
    • 1
  • Kunio Gonmori
    • 1
  • Osamu Suzuki
    • 1
  1. 1.Department of Legal MedicineHamamatsu University School of MedicineHamamatsuJapan
  2. 2.Environmental Nanotoxicology Section, Research Center for Environmental RiskNational Institute for Environmental StudiesTsukubaJapan
  3. 3.Department of Legal MedicineSt. Marianna University School of MedicineKawasakiJapan

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