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The fluorimetric determination of serum cortisol was improved1. using a pumpdevice to fill a special microcuvette avoiding the development of small bubbles,2. using a recording spectrofluorometer with optimal absorption (464 nm) and emission (522 nm), and3. allowing for maximal fluorescence of cortisol (80 min).Sensitivity (<1 µg cortisol/100 ml), accuracy, precision and specificity of the method are reported. Normal values of 9.00 a.m. serum cortisol (9.7–32.0 µg/100 ml) and of values before and after ACTH infusion tests were determined.For adrenal insufficiency (Addisons disease, total adrenalectomy, or after hypophysectomy) the 9.00 a.m. values of serum cortisol were generally satisfactory for diagnosis. In partial adrenal insufficiency ACTH infusion tests had to be performed.Very low levels of serum cortisol (2–4 µg/100 ml) were obtained, when patients with adrenal insufficiency were substituted with dexamethasone for three days, proving the specificity of the method.Determination of circadian rhythms of serum cortisol in patients with adrenal insufficiency on cortisol substitutive therapy in divided doses demonstrated cortisol levels far below the normal values during the carly morning hours. This situation should be improved by dividing the cortisol dose as follows: 6 a.m.: 10 mg, 10 a.m.: 5 mg, 2 p.m.: 5 mg and 8 p.m. or later: 10 mg cortisol.

Eine verbesserte fluorimetrische Cortisolbestimmung im Serum: Diagnostische Bedeutung und therapeutische Folgerungen bei NNR-Insuffizienz