Sensitive Determination of Gemcitabine Molecule by Novel Magnetic Solid-Phase Extraction Coupled with HPLC-DAD

Gemcitabine (GEM) is a widely employed antimetabolite with broad antineoplastic properties and is commonly administered in the treatment of pancreatic, breast, and ovarian cancers. In this study, a new sample preparation and quantification strategy was established to enable the trace-level determination of GEM. The approach integrates magnetic solid-phase extraction (MSPE) with subsequent HPLC–DAD analysis. Within the method, GEM is initially retained on a magnetic sorbent in a pH 5.0 buffer medium, followed by desorption into a reduced volume of ethanol to facilitate chromatographic measurement. Key experimental parameters particularly adsorption and desorption conditions as well as medium pH were systematically examined and optimized. Analytical performance characteristics, including the linear working range, enrichment factor, limit of detection, accuracy, and repeatability, were assessed using model solutions. Chromatographic detection of GEM was achieved at 277 nm under isocratic conditions composed of 60 % methanol, 25 % phosphate buffer at pH 6.0, and 15 % acetonitrile. Partial validation with model solutions demonstrated a detection limit of 3.43 ng mL⁻¹ and an enrichment factor of 100. Overall, the developed method offers high sensitivity, strong reproducibility, and reliable applicability for monitoring GEM at trace concentration levels.