Determination of the Stabilizer Sucrose in a Plasma-Derived Antithrombin Process Solution by Hydrophilic Interaction Chromatography with Evaporative Light-Scattering Detection

Hydrophilic interaction chromatography (HILIC) is used for the quantitation of sucrose in the range of 10–100 µg/mL. A poly-2-hydroxyethylaspartamide column is eluted with 25% water-75% acetonitrile, and evaporative light scattering is utilized for detection. A process sample of antithrombin (Atenativ) from Octapharma AB (Stockholm, Sweden) containing 20% sucrose is analyzed. The precision for this high-performance liquid chromatographic method is a percent relative standard deviation (%RSD) of 4, limit of detection (s/n = 3) of 1 µg/mL, and mean recovery of spiked samples of 101% (RSD% of 3, n = 6). Analysis time is 10 min/sample. Glucose, fructose, sodium citrate, sodium phosphate, Triton X-100, and tri-n-butyl phosphate do not interfere with the method.