Measurement of adrenal corticosteroids by high-performance liquid chromatography.
OBJECTIVE
To develop a high-performance liquid chromatography (HPLC) method for simultaneous measurement of adrenal corticosteroids important for the diagnosis of congenital adrenal hyperplasia.


METHODS
Blind comparison using convenience samples.


METHODS
Research laboratory at the Department of Medical Technology, Bowling Green State University.


METHODS
Referred samples for cortisol, 11-deoxycortisol (S), and 17-hydroxyprogesterone (OHP).  Stimulation condition unknown.


METHODS
Venous blood collected, processed to serum, and frozen. Adrenal steroids measured by radioimmunoassay (RIA) and by HPLC.


METHODS
Linearity, analytic sensitivity, accuracy, and precision were evaluated using recovery, coefficient of variation (c.v.), correlation coefficient (r), Student's t test, F test, and linear regression.


RESULTS
Calibration curves were linear to at least 50 mumol/L, and the low-end sensitivity was 10 nmol/L.  Analytic recovery ranged from 89.8% to 103.9%.  The CV was below 7.5% (n = 10) for all three steroids. Comparison to RIA yielded r values of 0.909 (n = 25), 0.932 (n = 10), and 0.983 (n = 10) for cortisol, S, and OHP, respectively.  Student's t-test results were 1.13 (p = 0.27), 0.10 (p = 0.93), and 0.48 (p = 0.64) for cortisol, S, and OHP, respectively.  F-test results were 1.14 (p = 0.75), 1.37 (p = 0.66), 2.39 (p = 0.21) for cortisol, S, and OHP, respectively.  Results for cortisol showed a negative bias of 97.0 nmol/L (3 mug/dL).  Linear regression analysis revealed proportional errors of -15.0% -7.0%, and +9.0%, for cortisol, S, and OHP, respectively.


CONCLUSIONS
(ABSTRACT TRUNCATED AT 250 WORDS)
Department of Medical Technology, Bowling Green State University, Ohio 43403.
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Harr
D000305:Adrenal Cortex Hormones / Q000032:analysis*; D000312:Adrenal Hyperplasia, Congenital / Q000175:diagnosis*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D011863:Radioimmunoassay; D012680:Sensitivity and Specificity