A new tool in metabolism research - Combination of OPLC on-line radioactivity detection with HPLC-RD technique

E. Mincsovics, B. Dalmadi Kiss, G. Morovján, K. Balogh Nemes, I. Klebovich

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

This paper reports a recently developed technique, coupling of optimum-performance laminar chromatography (OPLC) with on-line radioactivity detection (OPLC-RD). Sample preparation for purification of urinary metabolites of a 14C-labeled drug candidate was performed on octadecyl-modified silica solid-phase extraction columns. Optimum-performance laminar chromatography was conducted on aluminum-backed silica gel layers with butanol-acetic acid-water as mobile phase. Eluate radioactivity was detected by use of a flow-cell radioactivity detector equipped with solid scintillator, and eluate fractions containing radioactivity were collected. Strongly retained radioactive components were detected off-line after development by use of digital autoradiography (DAR). The isolated fractions containing metabolites were further purified by reversed-phase high-performance liquid chromatography coupled with radioactivity detection; this provided a second dimension for additional selectivity. Purified metabolite fractions were submitted to structure elucidation by spectroscopic methods. The novel on-line optimum-performance layer chromatography with radioactivity detection, combined with high-performance liquid chromatography with radioactivity detection and digital autoradiography techniques, the advantages and disadvantages of which are discussed in detail in this paper, provides a rapid, effective, and economic tool which can be applied to advantage in metabolism research.

Original languageEnglish
Pages (from-to)312-317
Number of pages6
JournalJournal of Planar Chromatography - Modern TLC
Volume14
Issue number5
Publication statusPublished - 2001

Fingerprint

Radioactivity
Chromatography
Metabolism
High Pressure Liquid Chromatography
Research
Metabolites
High performance liquid chromatography
Autoradiography
Butanols
Silica Gel
Solid Phase Extraction
Reverse-Phase Chromatography
Aluminum
Silicon Dioxide
Acetic Acid
Phosphors
Purification
Economics
Detectors
Water

Keywords

  • Digital autoradiography (DAR)
  • HPLC-radioactivity detection (HPLC-RD)
  • Hyphenated techniques
  • Metabolism study
  • On-line OPLC-radioactivity detection (OPLC-RD)
  • Optimum performance laminar chromatography (OPLC)
  • Overpressured layer chromatography (OPLC)

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

A new tool in metabolism research - Combination of OPLC on-line radioactivity detection with HPLC-RD technique. / Mincsovics, E.; Dalmadi Kiss, B.; Morovján, G.; Balogh Nemes, K.; Klebovich, I.

In: Journal of Planar Chromatography - Modern TLC, Vol. 14, No. 5, 2001, p. 312-317.

Research output: Contribution to journalArticle

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