Light-scattering and turbidimetric detection of silica colloids in size-exclusion chromatography |
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Authors: | Zarina Aspanut Tomomi Yamada Lee Wah Lim Toyohide Takeuchi |
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Institution: | (1) Department of Chemistry, Faculty of Engineering, Gifu University, 1-1 Yanagido, Gifu 501-1193, Japan |
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Abstract: | Silica colloids were separated by size-exclusion chromatography and monitored by fluorimetric and UV detection. In the former
means of detection, silica colloids were visualized by light-scattering. The signal intensity based on the light scattering
increased with increasing size of the silica colloids. The maximum intensity was observed at excitation wavelengths around
270–290nm. In UV detection, silica colloids were visualized based on turbidimetry, and the signal intensity also increased
with increasing size of the silica colloids and with decreasing detection wavelength. The signal intensities for both light-scattering
and turbidimetric detection were a linear function of the concentration of the silica colloids. The detection limit at S/N = 3 for 78-nm colloids was 0.06 ppm for light-scattering detection whereas the LOD was 2.3 ppm for UV detection. Effects
of mobile phase conditions and flow rate on resolution and peak shape were examined. Use of phosphate buffer allowed the separation
of silica colloids of different sizes in size-exclusion chromatography. |
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Keywords: | Silica colloids Light-scattering detection Turbidimetric detection Size-exclusion chromatography |
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