Determination of M/G ratio of propylene glycol alginate sodium sulfate by HPLC with pre-column derivatization

• A reliable HPLC method was developed for determination of the M/G ratio of PSS.
• A mild hydrolysis method with low uronic acid loss was developed.
• M/G ratio determined by this method was similar to the 1H NMR method.
• The M/G ratios of PSS samples from different manufacturers were compared.

A reliable high performance liquid chromatography with pre-column derivatization method was developed for the determination of the mannuronic acid (M)/guluronic acid (G) ratio of propylene glycol alginate sodium sulfate (PSS). The hydrolysis conditions of PSS were investigated by four degradation methods based on the degree of destruction of M and G, and the chromatographic separation conditions were also optimized. A satisfactory resolution of M and G was achieved with a KP-C18 column using 0.1 mol/L phosphate buffer (pH 7.0)-acetonitrile (83/17, v/v) as a mobile phase, after PSS was hydrolyzed with 0.1 mol/L sulfuric acid and labeled with 1-phenyl-3-methyl -5-pyrazolone. The M/G ratio of PSS determined by this method was in good accordance with that obtained by the 1H NMR method with a desulfurization strategy. Our method is rapid, sensitive, accurate and reproducible. The limit of detection was found to be 0.25 μg/mL for M and 0.40 μg/mL for G.