Fluorometric determination of plasma 11-hydroxycorticosteroids. II. Studies on the specificity of the method.

We have investigated the specificity of the fluorometric method we proposed [C/in. Chem. 19, 710 (1973)] for plasma or serum li-hydroxycorticosteroid determinations. The principal specific fluorogenic contaminants in the cortisol-containing extract of plasma, as detected by thin-layer chromatography and chemically, were triglycerides and total cholesterol. These contaminants contributed an average of 1.6 tg/dl to the total normal cortisol value. Fatty acids were also found, but did not fluoresce. Nonspecific serum fluorogens were quantitated at 1.3 ± 1.2 tg/dl. Cortisol, corticosterone, and fluorogenic contaminants represented an average of 62.0, 29.1, and 9.9%, respectively, of the total fluorometric plasma cortisol value obtained (expressed in terms of cortisol standard). A ratio of 8.3 ± 2.4 was found for cortisol/corticosterone when each component was determined in terms of its respective standard. Fluorescence scans of the plasma cortisol extract indicated cortisol to be the main component present, accompanied by minor fluorescent contaminants.