Development and clinical validation of HPLC method for precise quantification of paclitaxel in plasma from breast...

A simple, reproducible, and green high-performance liquid chromatography (HPLC) method was developed and validated for the precise quantification of paclitaxel in plasma following an intravenous dose in patients with breast cancer. The proposed method is developed to enhance efficiency, offering significant advantages including rapid analysis, one-step extraction, low solvent consumption, and thorough validation with extensive parameters. Chromatographic analysis was performed using Schimadzu HPLC with LC solution software. Plasma proteins were precipitated using acetonitrile for efficient extraction of paclitaxel. A separation was achieved on C18 column with the mobile phase being a combination of acetonitrile and water in the combination of 65:35 (v/v). The method exhibited excellent linearity (with a correlation coefficient of 0.9999) over the concentration range of 0.375 μg/mL to 8 μg/mL. The lower limit of quantification was found to be 0.375 μg/mL. The accuracy and precision of the intra-day (96.87-99.65%) and inter-day (98.3-99.9%) assays, comply with accepted bioanalytical criteria. The recovery of paclitaxel from plasma ranged between 96.97 and 100%. Stability studies indicated that paclitaxel remained stable after 24 hours at room temperature and 4 weeks at -20°C. AGREE, AGREEprep, ComplexGAPI, AMVI, AMGS, and ESA tools confirmed the environmental friendliness and greenness of the proposed method. Furthermore, the method was successfully utilized in the pharmacokinetic investigation of paclitaxel in breast cancer patients. The determined values of Cmax were found comparable with reported values, confirming the method's suitability for clinical pharmacokinetic study.