Reference ranges for gastric emptying time (GET), small intestinal transit time (SITT), and colonic transit time of 1.5-mm and 5-mm radiopaque markers in healthy cats fed a high-fiber meal were determined, and the influence of low-dose diazepam intravenous injection on the gastrointestinal transit of the markers was examined. The mean GETs and SITTs, and the mean residence times (MRTs) and geometric centers (GCs) of markers in the colon were determined. The effect of intravenous diazepam injection and marker size on these parameters was examined. Diazepam injection had no significant influence on gastrointestinal transit. The GETs of the 1.5-mm markers were significantly more rapid than those of the 5.0-mm markers. There were no significant differences between the SITTs or GCs of the 1.5-mm and 5.0-mm markers. Reference values were developed for GET, SITT, and colonic transit of radiopaque markers for cats fed a high-fiber meal. Diazepam injection had no effect on these parameters.