Measurement of total calcium by flash chronopotentiometry at polymer membrane ion-selective electrodes

Ionophore-based ion-selective electrodes are widely used for potentiometric electrolyte measurements, in which case they are known to detect the free ion activity. Total ion concentrations cannot be directly assessed by this methodology if the ion is predominantly present in a complexed form. We present here the direct measurement of total calcium using a calcium ion-selective electrode interrogated in a flash chronopotentiometric transduction mode. A high magnitude of cathodic current pulse is applied across a calcium ion-selective membrane containing the ionophore ETH 5234 but void of ion-exchanger to prevent spontaneous extraction. This induces a defined flux of calcium ions from the sample side to the membrane and results in the release of labile bound calcium and a concomitant depletion at the membrane surface at a critical current or time. This is observed as an inflection point on the potential-time curve and the square root of the transition time is linearly related to the total concentration in the sample. It is shown that the responses to solutions of labile calcium complexes of nitrilotriacetic acid (NTA) are in a good agreement with that of the same concentration of calcium chloride in saline solution with this protocol. Initial applications are aimed towards assaying extracellular calcium. Calcium binding to albumin is shown to be inconsequential with sample dilutions typical for clinical assays. Calcium calibration curves in real and artificial dilute serum are finally shown to correspond to that of calcium chloride, suggesting that the methodology is indeed capable of detecting total calcium under these conditions. The present membrane materials allow detection of up to over 0.5mM total calcium in serum, currently requiring such samples to be diluted about 5-fold. The slopes of the square root of time-concentration dependence for the calibrations of free calcium in a background of NaCl and total serum calcium were found to be 3.857 and 3.717 s(1/2)mM(-1), respectively, deviating by just 3.6%. The lower detection limit (3x SD) was calculated as 12 microM.