METHOD OF MEASURING NUMBER OF POLYSORBATE-80 WITH APPLICATION OF ALKALINE HYDROLYSIS OF SAMPLE WITH FOLLOWING HPLC

摘要

FIELD: technological processes.;SUBSTANCE: invention relates to methods for quantitatively determining polysorbate-80 in solutions of therapeutic proteins. Method for determining the concentration of polysorbate-80 in a sample containing a protein or a glycoprotein, comprising conducting an alkaline hydrolysis of samples, differs in that after carrying out alkaline hydrolysis, the sample is analyzed by high-performance liquid chromatography in isocratic mode for 13–30 minutes, at a phase flow rate of 0.02–5.0 ml/min, with a volume ratio of the components of the water-acetonitrile mobile phase in the range of 30/70 to 15/95, the column length is 30 to 300 mm, the sorbent pore size is 1.7 to 5.0 mcm, detection of the analyte is carried out in the UV region of the spectrum in the wavelength range 190–220 nm, and the reaction product is identified by chromatography of the analytical standard of oleic acid with recording the retention time of its peak.;EFFECT: method for fast high-performance liquid chromatography is proposed.;11 cl, 5 dwg, 6 tbl, 4 ex