Sera were analyzed for calcium ion activity on the Orion 99-20 and SS-20 systems. Samples from 84 healthy subjects were analyzed on both systems and, because of a change of electrode composition, 74 of them were reanalyzed after this change. Additional 89 were analyzed on only SS-20. The distribution of serum calcium ion activity was close to Gaussian. The difference between the 99-20 and SS-20 readings was 0.048+/-0.017 mmol/liter (range -0.01 to 0.08) before the electrode change and -0.085+/-0.027 mmol/liter (range -0.14 to 0.02) after. The reference ranges were 1.02--1.23 mmol/liter for 99-20 and 0.93--1.20 mmol/liter for SS-20 before the electrode change. The following characteristics may contribute to differences in the readings: higher temperature, higher flow rate, new calcium and reference electrodes, new liquid junction construction and a new pumping procedure. For 74 subjects the determination of serum calcium ion activity (99-20 system) was performed with a five year interval and the difference between the first and second measurements was -0.0046+/-0.0168 mmol/liter (mean +/-SD), ranging from -0.04 to 0.03 mmol/liter. This corresponded to an intraindividual variation of 1.5%. The corresponding variations for serum total calcium and corrected calcium were 4.8% and 3.3% respectively.