The matrix of SDS integrated with linear hydrophilic polymer for resolution of high- and low-molecular weight hyaluronic acids in MEKC

Abstract

Hyaluronic acid (HA), a multi-functional material, has a high dispersion in molecular weight, and the functions of HA are determined through the size. Nevertheless, hyaluronic acid mixtures are not easily separated due to their polydispersity. In this study, a capillary electrophoresis strategy was developed for resolution of different molecular-weight HA without enzymatic digestion. Here, hyaluronic acid mixtures with low molecular weight (380 kD; LHA) and high molecular weight (2180 kD; HHA) were successfully resolved by the SDS integrated with low molecular-weight polymer in capillary electrophoresis. By optimizing experimental conditions, the separation of LHA and HHA was completed within 14 min. The optimal conditions were as follows: the running buffer was 25 mM borate buffer (pH 9.75) containing 30 mM SDS and 10% polyethylene glycol (MW: 8000); applied voltage was 20 kV (detector at cathode side) and separation temperature was set at 25 °C. The data of method validation showed that calibration plots were linear (r ≥ 0.9977) over a range of 10-50 μg/mL for LHA, and 40-200 μg/mL for HHA. In the evaluation of precision and accuracy for this method, the RSD and RE values were all less than 4.2%. This fascinating technique was successfully applied to the quality control of cosmetic and pharmaceutical containing different ratios of LHA and HHA, and it was feasible for serving as a tool to quantitatively analyze different sizes of HA for clinical survey.

Keywords: HHA; Hyaluronic acids; LHA; MEKC; PEG sieving matrix; SDS micelles.