Investigation of Chromatographic Surface Viscosity Using Total Internal Reflection Fluorescence

Abstract

Total internal reflection fluorescence (TIRF) spectroscopy was employed to determine the viscosity of planar chromatographic surfaces. Various n-alkyldimethylchlorosilanes were used to modify the surface of a UV-quartz plate, so as to obtain a covalently attached monomeric C₁, C₃, C₈, or C₁₈ surface. TIRF anisotropy data of 1, 6-diphenylhexatriene (DPH) were collected on the unmodified and modified surfaces in the presence of overlaying solvents covering a range of viscosities. The results indicate that DPH adsorbs onto C₁ and C₃, but clearly exhibits partitioning with the C₁₈ phase. The C₈ surface data indicate intermediate behavior between adsorption and partitioning. On the basis of these preliminary data, the estimated microviscosity of a planar C¹⁸ surface is approximately 18 cP.

Keywords

Total internal reflection fluorescence Fluorescence anisotropy Chromatographic surface viscosity

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