Histamine release by compound 48/80 was substantially reduced in a time-dependent manner (maximum at 30 min) by pre-incubating mast cells in calcium-free medium at 37 degrees C but not at 2 degrees C. This effect was optimal at pH 7.0 to 7.5. 2. The re-introduction of calcium (0.1 to 3 mmol/l) restored histamine release to the control value; this effect was independent of temperature. 3. Strontium (1 to 30 mmol/l) partially reversed the effect of calcium deprivation but the same concentrations of barium and magnesium depressed histamine release even further. Magnesium (3 to 15 mmol/l) antagonized the effect of calcium replacement. 4. Results suggest that the level of cell-fixed calcium involved in compound 48/80-induced histamine release may be controlled by the combination of rapid passive influx and slow active efflux.