Separation, Characterization and quantification of Impurities and Identification of stress degradants of Cabazitaxel by RP-HPLC and LC-ESI-MS techniques

Shimadzu HPLC prominence coupled with MS Spectrometry (THERMO) with Sunfire C18 column (100x 4.6 mm, 3.5 pm particle size) was used for performing the chromatographic separation with 0.05% formic acid and acetonitrile mixture as mobile phase with flow rate 1.0 ml/min.The injection volume 10pL and wavelength was monitored at 220 nm). Shimadzu HPLC prominence coupled with MS Spectrometry (Thermo) with Sunfire C18 column (100 x 4.6 mm, 3.5 pm particle size) was used for the chromatographic study. 0.05% formic acid and acetonitrile mixture was chosen as mobile phase and the system was operated on gradient mode with flow rate 1.0 ml/min.The injection volume was 10pL and detection wavelength was 220 nm. Cabazitaxel API (Active pharmaceutical ingredient) and its seven impurities were procured from Dr. Reddy's Labs (Hyderabad, India) and used as it is without further purification.HPLC grade acetonitrile, formic acid, hydrochloric acid, sodium hydroxide and 30% w/v hydrogen peroxide were purchased from Merck and Milli-Q water from Millipore system was used.Cabazitaxel stock solution spiked with its seven impurities (I-VII) were prepared in acetonitrile and stored. Mass detector (Ion trap mass detector) with Electrospray ionization (both positive and negative modes) maintaining capillary temperature 250 °C (Spray/Source voltage (kV): 4.5; Sheath gas flow (Arb): 30.00; Aux/Sweep gas flow (Arb): 5.00).