New techniques for ion-selective measurements of ionized calcium in serum after pH adjustment of aerobically handled sera.

I report further experience in measuring ionized calcium (Ca2+) with the AMT Electron System and its serum standards and solid-state, dip, calcium-selective electrodes. With this system, serum pH can be adjusted with CO2 gas and Ca2+ and pH simultaneously measured; when 5.2% CO2 (40 mm pco2) is used for sample equilibration, the standard bicarbonate concentration is also provided. I measured serum Ca2+ as a function of pH between pH 7.0 and 9.0 and found the relationship to be reproducible, with no evidence of irreversible complexing of Ca2+. When the pH of aerobically exposed, mailed sera was restored to the original values, their values for Ca2+ were the same as for the fresh sera. Measurement of Ca2+ in routinely (aerobically) handled sera after pH restoration with CO2 gas was therefore validated, both samples from within an institution and mailed specimens. Standardization to pH 7.40 is recommended for routine measurements, is generally more accurate than use of heparin or quasianaerobic techniques, and is a practical approach. In patients with possible uncompensated acid-base disturbance (which may be indicated by an abnormal standard bicarbonate concentration if not suspected clinically), patient pH should be measured independently as part of the usual strict, anaerobic blood-gas-analysis procedures. Abnormal patient pH must be considered in the interpretation of Ca2+ results determined at pH 7.40 which are borderline or slightly abnormal; most accurately, Ca2+ may be measured in the separated sera at the previously determined patient pH value. Studies of aqueous solutions with the currently used Ca2+ electrodes showed a selectivity coefficient (the constant which relates the activity of an interfering ion to the activity of calcium that would contribute the same emf) KNa=0.0031 +/- 0.0003 (SE) and KMg=0.046 +/- 0.004 (SE). At physiological concentrations of Ca2+, physiologically encountered variation in Na+ is of no significance in resulting Ca2+, but extreme variation in Mg2+ may cause an error of approximately 1%.