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 micro g/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 microg/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.
Karlsson
Göran
G
Octapharma AB, SE-11275 Stockholm, Sweden.
Hinz
Ann-Charlotte
AC
Winge
Stefan
S
D000991:Antithrombins / Q000737:chemistry*; D001769:Blood; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D008027:Light; D015203:Reproducibility of Results; D012542:Scattering, Radiation; D012680:Sensitivity and Specificity; D012996:Solutions; D013395:Sucrose / Q000032:analysis*
D000991:Antithrombins; D012996:Solutions; D013395:Sucrose