Determination of hormones in human urine by ultra-high-performance liquid chromatography/triple-quadrupole mass spectrometry

Steroid hormones play a critical role in maintaining the homeostasis of human metabolism. Urine as a noninvasive sample has been extensively used in clinical diagnosis for hormones homeostasis. In this study, the simultaneous characterization of fourteen hormones in urine was performed based on ultra-high-performance liquid chromatography/electrospray ionization tandem mass spectrometry (UPHLC/ESI(+)-MS/MS) with multiple reaction monitoring in the positive ionization mode. The target hormones were cortisone, cortisol, 11-deoxycortisol, aldosterone, corticosterone, 11-deoxycorticosterone, progesterone, 17-OH-progesterone, pregnenolone, estrone, estradiol, estriol, testosterone and dehydreopiandrosterone. β-Glucuronidase/sulfatase deconjugation and liquid–liquid extraction (LLE) were conducted for the determination of urinary hormones (free + conjugated forms). The limits of detection (LODs) ranged from 0.2 ng/mL (11-deoxycortisol and testosterone) to 1 ng/mL (cortisone). The extraction recovery of the targeted compounds ranged from 87% to 127%, indicating sufficient extraction efficiency for the LLE process. Intraday precision was below 10% and the accuracy ranged from 84% to 122%. The profiling analysis of hormones in urine samples helps to understand the metabolic state of biological systems and can be employed as a diagnostic tool in diseases developed by endocrine-disrupted systems.