Ultra-high performance liquid chromatographic determination of levofloxacin in human plasma and prostate tissue with use of experimental design optimization procedures.

•An UHPLC-DAD method was developed for levofloxacin determination in human plasma and prostate samples of patients undergoing prostate biopsy.•The experimental design was successfully applied to optimize key parameters of extraction procedure, chromatographic separation and for robustness determination.•Validation studies demonstrated method’s sensitivity, reproducibility, precision and robustness in the range of linearity.•The method was successfully validated and applied to determine levofloxacine concentrations in 140 plasma and prostate samples.