Asymmetrical-Flow Field-Flow Fractionation with On-Line Multiangle Light Scattering Detection. 1. Application to Wormlike Chain Analysis of Weakly Stiff Polymer Chains

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Abstract
Four samples of hyaluronan in the sodium form, ranging in weight-average molecular weight, Mw, from 6.67 × 105 to 4.23 × 106 were investigated by asymmetrical-flow field-flow fractionation coupled to multiangle light scattering (FlFFF−MALS) in 0.2 M aqueous NaCl at 25 °C. Mw and z-average radii of gyration, RGz, obtained via FlFFF−MALS showed a good agreement with the results obtained by conventional static light scattering. Furthermore, the molecular weight dependence of the radius of gyration for sodium hyaluronan obtained via FlFFF−MALS was analyzed on the basis of the Kratky−Porod model for unperturbed wormlike chains combined with the Yamakawa theory for radius expansion factor, and a sufficiently good agreement was observed between the theoretical prediction and experimental data. These results show the potential usage of FlFFF−MALS regarding size separation and molecular characterization even for weakly stiff chains.