Development of a fast fiber based UV-Vis multiwavelength detector for an ultracentrifuge

Abstract

The advantages of simultaneously detecting multiple wavelengths in ultracentrifugation experiments are obvious, especially for interacting systems. In addition, the detection of the wavelength dependence of turbidity opens up the possibility to obtain independent information on the particle size in addition to the usual sedimentation coefficient distribution for colloidal systems. We therefore made an effort to develop a fast UV/Vis detector, which is able to simultaneously detect the range from 200-800 nm. This is possible by the use of a modern CCD chip based generation of UV-Vis spectrometers, which translates the dispersed white light onto a CCD chip, where each pixel corresponds to a particular wavelength. In addition to the simultaneous detection of a large number of wavelengths in the range 200-800 nm, also with non integer values, these spectrometers are very fast. Current typical spectrum scan times with the necessary scan quality in the ultracentrifuge are in the range of 100 ms but this time can be significantly shortened down to 3 ms for higher light intensities and even down to 10 μs for a new generation of CCD chip based spectrometers. The introduction of a fiber based UV-Vis optics into a preparative XL-80K ultracentrifuge with the associated hardware developments will be described as a first generation prototype. In this study, we use a wavelength dependent optical lens system instead of the necessary but more complex wavelength independent mirror optical system for a first check on possibilities and limitations of the optical system. First examples for biopolymers and latexes will be presented and compared to those obtained in the commercial XL-A ultracentrifuge. Already the fast detection enables completely new possibilities like the determination of a particle size distribution in a few minutes. Multiwavelength detection at constant position in dependence of time will be demonstrated, which is an important mode for the use of speed profiles for very polydisperse samples. Also, the use of radial multiwavelength scans will be demonstrated producing a three dimensional data space for monitoring the sedimentation via radial scans with multiwavelength detection. However, despite the advantages, the current problems with the detector will also be discussed including the main problem that much intensity is lost in the important UV range as a result of fiber coupling and bending. © Springer-Verlag Berlin Heidelberg 2006.