Flow field‐flow fractionation and multiangle light scattering for ultrahigh molecular weight sodium hyaluronate characterization

Abstract

This review describes the utility of flow field-flow fractionation coupled with multiangle light scattering and differential refractive index (FlFFF-MALS-DRI) detection methods for the separation of ultrahigh molecular weight sodium hyaluronate (NaHA) materials and for the characterization of molecular weight distribution as well as structural determination. The sodium salt of hyaluronic acid (HA), NaHA, is a water-soluble polysaccharide with a broad range of molecular weights (10(5) -10(8) ) found in various naturally occurring fluids and tissues. Basic principles of FlFFF-MALS using field programming for the separation of the degraded products of NaHA prepared by treating raw materials with depolymerization or degradation processes such as membrane filtration, enzymatic degradation, ultrasonic degradation, alkaline reaction, irradiation by γ-rays, and thermal treatment for the development of pharmaceutical applications are introduced. Changes in molecular weight distribution and conformation of NaHA materials due to external stimuli are also discussed.

Detailed characterization of hyaluronan using aqueous size exclusion chromatography with triple detection and multiangle light scattering detection

“…For the three lots of HA tested, the values of the conformation plot slope are 0.6, which is consistent with HA having a random coil conformation in the aqueous solution tested. This result is in good agreement with the general findings in the literature reports 15, 16, 27, 37…”

“…This will help ensure that a certain molecular weight of HA is maintained over the shelf life of the product, which will help enhance comfort. Size exclusion chromatography (SEC) and flow field–flow fractionation with various advanced detection methods have been proved to be effective tools to evaluate the solution properties of HA and other biomaterials to ensure the quality of the products for specific claims 15, 16…”

Molecular characterization of branched polysaccharides from Tremella fuciformis by asymmetrical flow field‐flow fractionation and size exclusion chromatography

“…Asymmetrical flow field‐flow fractionation (AF4) coupled with MALLS has been applied for the molecular characterization of synthetic polymers , hyaluronic acid , as well as starch, cellulose, gums, alginate, and cereal β‐glucan . Compared to SEC, AF4 has some important advantages including (1) no stationary phase is required, so shear degradation is strongly minimized and abnormal elution effect can be avoided, (2) the exclusion limit for AF4 is at least two orders higher than that for SEC, (3) the low surface area of the accumulation wall in AF4 avoids unwanted adsorption and secondary separation effects .…”

Advanced fractionation methods for the microstructure analysis of complex polymers

“…Asymmetric flow FFF is driven by the normal diffusion coefficient as a function of molecular size and, therefore, is primarily used for the fractionation of complex samples according to molecular size . In ThF3, a temperature gradient is applied perpendicular to the axial carrier liquid flow, which is achieved by combining a hot and a cold plate as FFF channel walls.…”

“…[28][29][30][31][32][33] In ThF3, a temperature gradient is applied perpendicular to the axial carrier liquid flow, which is achieved by combining a hot and a cold plate as FFF channel walls. The analyte molecules are driven to the cold wall (accumulation wall) by their interaction with the applied temperature gradient.…”