Evaluation of commercial silica-gel HPTLC plates for quantitative fluorescence analysis

Yuanling Liang, Mark E. Baker, Daniel A. Gilmore, M. Bonner Denton

Research output: Contribution to journalArticle

12 Scopus citations

Abstract

Several commercially available high-performance thin-layer chromatographic (HPTLC) plates were evaluated for quantitative fluorescence imaging analysis using scientific charge-coupled device (CCD) detection. This investigation examined the effects on the observed background intensity as it relates to: the technique used for precleaning the plates, the fluorescence from the plate backing material, the presence of inhomogeneous fluorescence 'impurities', and the thickness of the glass backing material and silica layer. The sensitivity and linear dynamic range were found to be limited primarily by the intensity of the fluorescence background from the plate rather than by the detector or the excitation source. The results show that plate precleaning is a crucial step in providing for a correctable fluorescence background sufficient for obtaining reliable quantitative results. The influence of particle size on sensitivity was negligible compared with the florescence background from the plates. With plate precleaning, detection limits of approximately 100 fg were obtained for rhodamine 6G with an intraimage linear dynamic range of four orders of magnitude when utilizing scientific CCD fluorescence detection.

Original languageEnglish (US)
Pages (from-to)247-253
Number of pages7
JournalJournal of Planar Chromatography - Modern TLC
Volume9
Issue number4
StatePublished - Oct 4 1996

Keywords

  • CCD detection
  • Imaging HPTLC
  • Plate characterization
  • Quantitative fluorescence analysis

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Clinical Biochemistry

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