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Clinical Chemistry 39: 2518-2521, 1993;
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Clinical Chemistry, Vol 39, 2518-2521, Copyright © 1993 by American Association for Clinical Chemistry

Free cortisol in serum assayed by temperature-controlled ultrafiltration before fluorescence polarization immunoassay

EG Lentjes, F Romijn, RJ Maassen, L de Graaf, P Gautier and AJ Moolenaar
Department of Clinical Chemistry, University Hospital Leiden, The Netherlands.

A method is described for a temperature-controlled ultrafiltration procedure to measure free cortisol in serum. A special thermometer with a sensor was developed, measuring the temperature directly in the ultrafiltration device. The sensor is screwed on the axis of the centrifuge rotor, and the centrifuge is placed in a temperature- controlled box so that the temperature of the sample is kept at 37 degrees C +/- 0.1 degrees C. The overall CV of the free cortisol assay ranges from 2.2% to 11.4%, of which the ultrafiltration contributes only 2.2-3.6%. Increasing amounts of cortisol-binding protein, as found in women using estrogen-containing oral contraceptives, have minor but significant effects on the free cortisol concentrations in serum. Serum free cortisol concentrations in a reference population (n = 114; central 95 percentiles) were 12-43 nmol/L (4-9.5% of total cortisol); in the group of the oral-contraceptive users (n = 27), the reference interval was 11-53 nmol/L (1.5-4.5%).


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