Serum ionized calcium: reference interval estimation and blood collection condictions

INTRODUCTION: The ionized serum calcium determination has become frequent replacing the total calcium with advantages: it is the physiologically active fraction, and the availability of robust methodology with favorable cost/benefice ratio. The use of this methodology implies in reference interval estimation. OBJECTIVE: To estimate the reference interval for serum ionized calcium, and to evaluate interferences in tourniquet time application, and interferences in sample refrigeration before analysis. MATERIAL AND METHOD: to estimate the reference interval we included the results of 11,320 consecutive calcium ionized determinations accomplished from January 2000 to November 2002; in order to evaluate the effect of sample refrigeration, 16 samples were collected in duplicate, so that one tube was placed in ice bath and the other was maintained in room temperature. To evaluate the effect of tourniquet application time, we collected blood samples from one arm of 6 normal subjects, immediately after, and from the other arm, after 3 minutes of the tourniquet application. The blood was collected in evacuated tubes with gel separator and centrifuged up to 30 minutes after collection. All determinations were performed up to 4 hours after the centrifugation by ion-selective electrode. RESULTS: regarding to the central 95% data distribution, the inferior and superior limits were, respectively, 1.11 (confidence interval of 90%: 1.1 to 1.11) and 1.4mmol/l (confidence interval of 90%: 1.39 to 1.41). No significant differences were detected between results with and without refrigeration and between samples with less than 1 and after 3 minutes of tourniquet application.

Ionized calcium; Reference values; Pre-analytical variation

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