On the coupling of fluorescence line-narrowing spectroscopy and poly(ethylene)imine-cellulose thin-layer chromatography

Autor: Cees Gooijer, Steven J. Kok, Nel H. Velthorst, U.A.Th. Brinkman, R. Evertsen
Přispěvatelé: BioAnalytical Chemistry, Chemistry and Pharmaceutical Sciences, Chemical Engineering and Chemistry
Rok vydání: 2000
Předmět:
Zdroj: Analytica Chimica Acta, 405, 1-7. Elsevier
Analytica Chimica Acta, 405(1-2), 1-7. Elsevier
Kok, S J, Evertsen, R, Velthorst, N H, Brinkman, U A T & Gooijer, C 2000, ' On the coupling of fluorescence line-narrowing spectroscopy and poly(ethylene)imine-cellulose thin-layer chromatography ', Analytica Chimica Acta, vol. 405, pp. 1-7 . https://doi.org/10.1016/S0003-2670(99)00698-4
ISSN: 0003-2670
Popis: The feasibility of a direct coupling of the low-temperature, high-resolution, fluorescence line-narrowing spectroscopy (FLNS) technique and the autoradiographic 32 P -post-labelling technique for DNA adducts is explored. FLNS has been used to identify analytes on various types of thin-layer chromatography (TLC) plates, but not on poly(ethylene)imine-cellulose (PEI-cellulose) TLC sheets involved in the 32 P -post-labelling technique which includes a two-dimensional separation. PEI-cellulose was found to exhibit a high fluorescence background. Its fluorescence excitation and emission characteristics were studied in detail and the signal-to-noise ratios of the FLNS spectra were optimized using background subtraction, time-resolved detection and a new synchronous scanning procedure. Benzo[a]pyrene (B[a]P) tetrols and a standard B[a]P–DNA adduct were used as model compounds. Limits of detection were in the low-picomole range. The technique was applied for the identification of spots extracted from PEI-cellulose TLC sheets obtained after a 32 P -post-labelling procedure of DNA containing B[a]P adducts at a level of two adducts per hundred bases.
Databáze: OpenAIRE
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