Abstract A new automatic device has been developed for the determination of the calcium content in biological fluids. The method is based on a fluorometric titration technique, in which calcium is titrated with ethyleneglycolbis(2-aminoethylether)-N, N,N′,N′-tetraacetic acid (EGTA). The end-point of the titration is indicated by quenching of the fluorescence of a calcium-calcein complex, in which calcein is the fluorescein-complexone indicator. To obtain proper results we found it necessary to use a 1 molar potassium hydroxide solution as titration medium. The results are printed out directly in concentration units (mmol/l). In order to carry out these determinations, several electronic and mechanical devices, including a sample-changer, titration cabinet with built-in fluorometer, control unit and electronic titrator, were developed. Together they form the new calcium automatic device of which several qualities such as precision, recovery and correlation have been studied. The disturbing influence of bilirubin has been minimized by using an excitation wavelength of 492 nm, at which there is nearly no absorption by bilirubin. At a concentration of bilirubin of as much as 500 μmol/l, the recovery of calcium is still about 96%.