COMPARISON OF CORTISON LEVELS DURING AN ACTH STIMULATION TEST USING TANDEM MASS

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This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The purpose of this study is to utilize tandem mass spectrometry, a novel analytical method, to measure serum cortisol levels following ACTH stimulation testing in patients who are normal and those suspected of having adrenal insufficiency. These determinations will be compared to the results obtained using more standard immunoassays in order to determine whether MS/MS measurement of cortisol will yield similar, but more accurate, values than standard immunoassay measurement of cortisol. A second goal will be to measure other steroid hormones (estradiol, total and unconjugated testosterone, DHEA, DHEA-S, androstenedione, 17-OH-progesterone, progesterone, cortisol, 11-deoxycortisol, total and uncongugated estriol, estrone, 2- OH -estrone, 16-OH- estrone, aldosterone and allopregnanolone) during an ACTH stimulation test. ACTH is known to increase the production of cortisol and aldosterone, but whether ACTH also stimulates other adrenal steroid production, and whether such measures might provide additional useful data in the evaluation of adrenal function, has not been studied. SPECIFIC AIMS: The primary purpose of this study is to compare measurements of cortisol using tandem mass spectrometry (MS/MS) (an experimental method) with simultaneously performed immunoassay (IA) (the standard method) during an ACTH stimulation test. A secondary goal will be to measure other steroid hormones (estradiol, total and unconjugated testosterone, DHEA, DHEA-S, androstenedione, 17-OH-progesterone, progesterone, cortisol, 11-deoxycortisol, total and uncongugated estriol, estrone, 2- OH -estrone, 16-OH- estrone, aldosterone and allopregnanolone) during an ACTH stimulation test. ACTH is known to increase the production of cortisol and aldoseterone, but it has not been well studied whether ACTH stimulates other adrenal steroid production. The expected benefits of measurement of cortisol with MS/MS over measurement of cortisol with IA are: 1) IA lacks specificity for all steroids including cortisol. 2) MS/MS measurements can potentially be performed more quickly (about 5 minutes by MS/MS versus about 30 minutes by IA). 3) MS/MS may be more accurate than IA. 4) MS/MS may be able to separate measurement of cortisol from therapeutic steroids, which would allow an ACTH stimulation test to be performed while someone is taking prednisone or other therapeutic steroids. The results of the MS/MS assays will not be used for clinical decision-making, but will be compared with the results of simultaneously performed IA. Hypothesis: Our hypothesis is that MS/MS measurement of cortisol will yield similar but more accurate values than IA measurement of cortisol.