Validation of an SEC-HPLC method for the analysis of rhG-CSF in pharmaceutical formulations

Granulocyte colony-stimulating factor (G-CSF) is a hematopoietic cytokine produced by recombinant DNA technology and used clinically to treat neutropenia. An isocratic high performance liquid chromatography procedure was developed for the assay of filgrastim in pharmaceutical formulations. HPLC separation was carried out by size-exclusion chromatography on a TSK gel G2000 SW column (60 cm x 7.5 mm I.D.). The mobile phase was composed of phosphoric acid (pH 2.5; 0.1 M), run at a flow rate of 1.0 mL min(-1) and with UV detection at 214 urn. Method validation investigated parameters such as the range, linearity (r(2) = 0.9998), precision, accuracy, and robustness; the method yielded good results with a quantitation limit of 45 mug mL(-1) and a detection limit of 12 mug mL(-1). The results demonstrate the validity of the SEC-HPLC method for the analysis of filgrastim.