Characterisation of locust bean gum with asymmetric flow field-flow fractionation (AF4) and light scattering

We present a detailed characterisation of locust bean gum (LBG), an industrially significant galactomannan, utilising asymmetric flow field-flow fractionation (AF4) and light scattering. Molecular weight and size determination of galactomannans is complicated by their tendency to aggregate, even in dilute solutions; AF4 allows us to confirm the presence of aggregates, separate these from well-dispersed polymer, and characterise both fractions. For the dispersed polymer, we find M_w=9.2×10⁵ g mol^-1 and R_g,z=82.1 nm; the distribution follows Flory scaling (R_g∼M^ν) with ν∼ 0.63, indicating good solvent conditions. The aggregate fraction exhibited radii of up to 1000 nm and masses of up to 3×10¹⁰ g mol^-1. Furthermore, we demonstrate how both fractions are influenced by changes to filtration procedure and solvent conditions. Notably, a 200 nm nylon membrane effectively removes the aggregated fraction; we present a concentration-dependent investigation of solutions following this protocol, using static and dynamic light scattering, which reveals additional weak aggregation in these unfractionated samples. Overall, we demonstrate that AF4 is highly suited to LBG characterisation, providing structural information for both well-dispersed and aggregated fractions, and expect the methods employed to apply similarly to other galactomannans and associating polymer systems.