The Determination of Hyaluronan Molecular Weight Distribution by Means of High-Performance Size Exclusion Chromatography

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Abstract
The high-performance liquid chromatographic (HPLC) method relevant to the size exclusion chromatographic mode (SEC) used for the distribution analysis of high-molecular-weight hyaluronans is described. The HPLC fillings used, to which was applied a phosphate buffer (50 mM, pH 7.8) effluent, consisted of a cross-linked hydroxyethylmethacrylate derivative gel (HEMA-BIO), which is commercially available, and a highly porous aminopropyl-silica sorbent (SG-10-6000-NH2), which is not commercially available. For reliable hyaluronans a high-molecular-weight distribution analysis is necessary involving SEC chromatographic columns which must have an extremely high exclusion limit (SG-10-6000-NH2). Calibration of these columns should be done using hyaluronate reference material.

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