Micro-analytical identification, by NMR, of elevated steroid in urine of Cushing's syndrome patients.

We previously reported that the daily urinary unidentified ketosteroid glucuronide(US-G) level in patients with Cushing’s syndrome was much higher than that in the healthy subjects. Furthermore, urine samples from patients with Cushing’s syndrome, including those with pituitary adenoma and adrenal adenoma, yielded almost the same high excretion levels, despite the different sites of the adenomas. We extracted US obtained by hydrolysis of US-G in urine of patients with Cushing’s syndrome, purified it, and analyzed its chemical structure. Molecular weight and molecular formula were analyzed by MS spectrometry, and the chemical structure was analyzed by NMR spectrometry, utilizing small quantities of refined US. The substance has a molecular weight of 304Da, a molecular formula of C19H28O3, and its chemical structure is 3α, 11β-dihydroxyandrost-4-en-17-one.