Procedure, Results & Discussion
High molar mass samples require lower flow-rates and concentrations in GPC/SEC. The GPC/SEC conditions and columns have been optimized for high molar mass hyaluronic acid with respect to loading, flow-rate and column particle size and porosity. For sample preparation the water content of approx. 12 % has been taken into account. Due to the lack of high molar mass calibration standards the use of light scattering detection is recommended. This allows also to measure true molar masses. A refractive index increment (dn/dc) for hyaluronic acid of 0.165 has been used to evaluate the light scattering data.1 Figure 1 shows the slice concentration measured by the RI detector as well as the on-line measured molar mass. Sample recovery (compare Conc. Calculation vs. Given) was nearly 100% showing that all material eluted from the column.
1 Lavrenko, Linow, Gornitz in Analytical Ultracentrifugation in Biochemistry and Polymer Science 517-531 Royal Society of Chemistry, Cambridge (1992)
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