Bencynonate is a newly developed anticholinergic drug. After a dose (2 mg/person or 4 mg/person) of bencynonate, the circulating concentrations are so low that therapeutic drug monitoring studies have previously been impossible. Radioisotope assay and radioaccepter assay had been used in rat experiments, but the detections limits of these methods were not sensitive enough for the therapeutic drug monitoring. Further, the two methods were unsuitable in the human. For evaluation of the pharmacokinetics and the relative bioavailability of bencynonate in man, a gas chromatographic--mass spectrometric method for quantitative analysis of bencynonate in human plasma was developed. Deuterated bencynonate served as the internal standard and selected-ion monitoring of the fragments of bencynonate and internal standard permitted the quantitation of bencynonate down to 25 pg.ml-1 of plasma. The linearity was in the range from 25 pg.ml-1 to 3 ng.ml-1 of bencynonate plasma concentration. At 0.25 ng.ml-1 level the recovery and relative standard deviation of variation are 54.3% and 19.1%, respectively. Application of the method to clinical studies gave the results of pharmacokinetics and relative bioavailability of bencynonate in man.