Abstract This study investigates matrix effects on a molecularly imprinted solid-phase extraction (MISPE) method developed for the clean-up of diphenyl phosphate (a hydrolysis product of the commonly used flame retardant and plasticizer, triphenyl phosphate) in urine samples. The influence of potentially interfering compounds that naturally occur in urine was examined with respect to extraction recovery, repeatability and selectivity. The components tested were NaCl, urea, creatinine and hippuric acid. The imprinted polymer was prepared using 2-vinylpyridine as the functional monomer, ethylene glycol dimethacrylate as crosslinker and a structural analogue of the analyte as the template molecule. The recovery of diphenyl phosphate from water standards was over 90% using MISPE, compared to less than 25% using a non-imprinted SPE (NISPE) counterpart. The selectivity of MISPE compared to NISPE was achieved in a wash step with a basic modifier in methanol. The recovery and repeatability of the MISPE method were affected most by NaCl in the tested concentrations, while urea, creatinine and hippuric acid had no significant influence. NaCl most likely weakens the binding during the loading of the sample. This effect could be suppressed by diluting the sample with a citrate buffer at pH 4.0.