Optimization of a HPLC–UV bioanalytical method using Box–Behnken design to determine the oral pharmacokinetics of neratinib maleate in Wistar rats

Abstract

AbstractThe current research work reports the development and validation of a sensitive, robust and reproducible bioanalytical method for quantifying neratinib maleate in rat plasma. More than 85% of the drug was extracted from the plasma samples by protein precipitation. The method was optimized using Box–Behnken design, a response surface method. The effect of three critical factors, viz., the pH of the buffer (X1), the aqueous phase proportion in the mobile phase (X2) and the mobile phase flow rate (X3), was studied on two response variables, retention time (Y1) and United States Pharmacopoeia (USP) width (Y2). With the highest overall desirability function value of 0.943, the obtained optimized method conditions were: X1 = 2.4 ± 0.1; X2 = 66.7 ml, and X3 = 0.85 ml/min. Under the optimized conditions, the values of Y1 and Y2 for a sample containing 1 ppm of the drug were found to be 14.1 min and 0.50 ± 0.003, respectively. Single‐dose intravenous bolus (7.5 mg/kg) and oral (15 mg/kg) pharmacokinetic studies were performed to determine the absolute bioavailability of the drug. The optimized bioanalytical method was sensitive enough to capture 95% of the drug eliminated from the body. The absolute oral bioavailability of the drug was 49.30%.