期刊
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
卷 407, 期 5, 页码 1327-1334出版社
SPRINGER HEIDELBERG
DOI: 10.1007/s00216-014-8379-2
关键词
Sodium hyaluronate; Flow field-flow fractionation; Multiangle light scattering; FlFFF-MALS; Ionic strength effect
资金
- National Research Foundation (NRF) - Korea government [NRF-2010-0014046]
This study describes the effect of ionic strength on the molecular structure of hyaluronic acid (HA) in an aqueous solution using flow field-flow fractionation and multiangle light scattering (FlFFF-MALS). Sodium salts of HA (NaHA) raw materials (similar to 2x10(6) Da) dispersed in different concentrations of NaCl prepared by repeated dilution/ ultrafiltration procedures were examined in order to study conformational changes in terms of the relationship between the radius of gyration and molecular weight (MW) and molecular weight distribution (MWD) of NaHA in solution. This was achieved by varying the ionic strength of the carrier solution used in a frit-inlet asymmetrical FlFFF (FIAF4) channel. Experiments showed that the average MW of NaHA increased as the ionic strength of the NaHA solution decreased due to enhanced entanglement or aggregation of HA molecules. Relatively large molecules (greater than similar to 5 MDa) did not show a large increase in RMS radius value as the NaCl concentration decreased. Conversely, smaller species showed larger changes, suggesting molecular expansion at lower ionic strengths. When the ionic strength of the FlFFF carrier solution was decreased, the HA species in a salt-rich solution (0.2 M NaCl) underwent rapid molecular aggregation during FlFFF separation. However, when salt-depleted HA samples (I=4.66 similar to 0.38 mM) were analyzed with FFF carrier solutions of a high ionic strength, the changes in both molecular structure and size were somewhat reversible, although there was a delay in correction of the molecular structure.
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