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HENRY J. RUDER, ROBERT L. GUY, MORTIMER B. LIPSETT, A Radioimmunoassay for Cortisol in Plasma and Urine, The Journal of Clinical Endocrinology & Metabolism, Volume 35, Issue 2, 1 August 1972, Pages 219–224, https://doi.org/10.1210/jcem-35-2-219
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ABSTRACT
A radioligand assay for the measurement of cortisol in plasma and urine has been developed. Antibodies raised against cortisol-21-hemisuccinate conjugated to bovine serum albumin had a high affinity for cortisol (K = 2 × 109 1/mole). A standard curve was established with a useful range from 0.10–4.0 ng. Plasma cortisol is readily measured by directly assaying small aliquots of a methylene chloride extract of plasma. Urine free cortisol was assayed after a single TLC of a methylene chloride extract of urine. Intraassay variability of both urine and plasma cortisol determinations was 10%; interassay variability was 20–26%. This method is particularly useful for measurement of 24-hr urine free cortisol.