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Light-scattering and turbidimetric detection of silica colloids in size-exclusion chromatography

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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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Acknowledgment

The authors would like to thank Tosoh Corporation, Tokyo, Japan, for their kind supply of the SEC separation columns. One of the authors (Z. Aspanut) would also like to thank the Public Service Department, Malaysia Government, for the kind offer of financial support.

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Authors and Affiliations

  1. Department of Chemistry, Faculty of Engineering, Gifu University, 1-1 Yanagido, Gifu, 501-1193, Japan

    Zarina Aspanut, Tomomi Yamada, Lee Wah Lim & Toyohide Takeuchi

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  1. Zarina Aspanut
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  2. Tomomi Yamada
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  3. Lee Wah Lim
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  4. Toyohide Takeuchi
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Correspondence to Toyohide Takeuchi.

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Aspanut, Z., Yamada, T., Lim, L.W. et al. Light-scattering and turbidimetric detection of silica colloids in size-exclusion chromatography. Anal Bioanal Chem 391, 353–359 (2008). https://doi.org/10.1007/s00216-008-2017-9

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  • DOI: https://doi.org/10.1007/s00216-008-2017-9

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