An FPSE-HPLC-PDA method for the determination of anticancer drugs in human whole blood, plasma, and urine

The present work describes a rapid, sensitive and specific method for quantifying residues of aromatase inhibitors including as anastrozole, letrozole and exemestane by high performance liquid chromatography (HPLC) in human plasma, whole blood and urine samples previously extracted by fabric phase sorptive extraction (FPSE) method. The proposed HPLC method, due to its ability to screen and quantify the three drugs concurrently, is very innovative since in literature there are only methods capable of monitoring a single inhibitor. These drugs were resolved using a Luna C18 column (250 mm × 4.6 mm; 5 m particle size) at room temperature (+25°C) with a mobile phases consisting of acetonitrile and phosphate buffer 40 mM (pH=3.5) in gradient elution mode within 20 minutes. The flow rate was set at 1mL/min. The analytical method was optimized and validated in the range from 0.025 µg/mL to 10 µg/mL for human plasma and urine; and in the range from 0.1 µg/mL to 10 µg/mL for human whole blood. Propyl-p-hydroxybenzoate was used as the internal standard (IS). Weighted-matrix matched standard calibration curves showed a good linearity up to a concentration of 10 g/mL. The intra- and inter-day accuracy values (precision and trueness) were found in the range from -9.21% to 12.8%. Finally, the performances of the validated FPSE-HPLC-DAD method was applied to real samples. To the best of our knowledge, this is the first FPSE extraction procedure applied to whole blood, plasma, and urine samples for the simultaneous determination of antitumor drugs possessing a wide range of logKow values (spanning from 2.31 for anastrozole, to 1.86 for letrozole, and 2.67 for exemestane) and could be easily used as a robust, rapid and green protocol for clinical and pharmaceutical applications.