Effect of EDTA on measurement of cortisol and thyroxine by chemiluminescent enzyme immunoassay in dogs.

The addition of ethylenediamine tetra-acetic acid (EDTA) to serum can affect the measurement of cortisol by chemiluminescent enzyme immunoassay (CEIA); addition of magnesium chloride (MgCl2) may reverse the effects. However, similar characteristics for thyroxine (T4) measurement are unknown. We measured cortisol and T4 in paired EDTA-anticoagulated plasma and serum samples from 50 dogs. Additionally, both hormones were measured in 15 samples of each type after the addition of MgCl2. Samples were collected under routine clinical conditions; therefore, specific EDTA concentrations in plasma samples were unknown. Cortisol and T4 values were significantly different comparing plasma and serum samples in the absence of MgCl2. For cortisol and T4, EDTA-plasma concentrations were 51.2% and 43.7% higher than serum, respectively (p < 0.001 for both). The addition of MgCl2 to plasma significantly decreased the measured cortisol concentrations (p < 0.001) but not T4 (p = 0.44). After addition of MgCl2, cortisol concentrations in EDTA-plasma were no longer significantly different from serum, whereas T4 concentrations in EDTA-plasma remained significantly different from serum. In the clinical setting in which tubes may be underfilled, use of EDTA-plasma significantly increases the measured concentration of cortisol and T4 obtained by CEIA. Addition of MgCl2 to EDTA-plasma can overcome the effects of EDTA when measuring cortisol, but not T4. Thus, T4 should not be measured in EDTA-plasma.