Low‐angle laser light scattering–aqueous size exclusion chromatography of polysaccharides: Molecular weight distribution and polymer branching determination

A low‐angle laser light scattering detector (LALLS) used with size exclusion chromatography (SEC/LALLS) has been applied for the determination of molecular weight, molecular weight distribution (MWD), and degree of branching of polysaccharides in 0.5 N NaOH aqueous solution. Data from both detectors [differential refractive index (DRI) and LALLS] are used to calculate the absolute molecular weight at each point in a sample chromatogram. The correct average molecular weight and MWD can be obtained without calibration methods used in conventional SEC. As a consequence of this technique, Mark—Houwink coefficients can be predicted from a single broad‐distribution, homopolymer without recourse to time‐consuming fractionation methods. Moreover, the hydrodynamic volume separation mechanism of SEC can be exploited with the SEC/LALLS method to gain information about polymer branching. In the studies described in this paper, SEC/LALLS has been employed to obtain data about the branching parameters g v and g M for samples of amylose, amylopectin, starch, and glycogen. For three homopolymers (amylose, amylopectin, and glycogen), branching frequency (as measured by chemical means), and the branching parameters ( g v and g M ) are inversely related. This trend is consistent with theoretical predictions. For starch, a nonhomogeneous branching distribution is observed as a function of molecular weight.