The method for simultaneous determination of nifedipine (NIF) and lidocaine (LID) in human plasma by one-step sample preparation has been developed for the first time. Due to the photosensitivity of nifedipine and its low plasma concentrations a precise and reliable method was required. The method involved liquid–liquid extraction (methyl tert-butyl ether, MTBE), and 10μL of the resulting sample was analyzed by HPLC–MS/MS. Chromatographic separation was achieved on an YMC-Triart C18 HPLC column (100×2.0mm; S‐5μm 12nm). The mobile phase was methanol:water, 60:40 (v/v) and contained 0.15% acetic acid. The linearity of the method was established in the concentration ranges of 0.5–50ng/mL for NIF and 1.0–500ng/mL for LID. Photodestruction of NIF under ambient light was evaluated. The validated method was successfully applied to analyze human plasma samples after rectal application of the drug (1g) containing 2.0% LID and 0.3% NIF.