Simultaneous Determination of Steroids and Glucocorticoids in Human Urine by Ultra-High-Performance Liquid Chromatography – Tandem Mass Spectrometry (UPLC–MS/MS)

Abstract

Prostate cancer is a hormone-dependent disease and therefore androgens play a critical role in its development. For both research on clinical conditions as well as diagnostic and monitoring purposes, accurate measurement of steroid hormones is essential. This study aimed to develop a sensitive and selective method for simultaneously detecting nine sex steroids and two glucocorticoids in human urine and to evaluate the alterations of urinary endogenous steroid hormones induced by prostate cancer. A total of 97 Chinese subjects were randomly recruited for this study, including 66 patients with prostate cancer and 31 healthy controls. The concentrations of steroid hormones, including dehydroepiandrosterone, dehydroepiandrosterone sulfate (DHEAS), androstenedione, testosterone, dihydrotestosterone (DHT), estrone, estradiol, estriol (E1), progesterone, cortisol (F), and cortisone (E), were simultaneously determined by ultra-high-performance liquid chromatography – tandem mass spectrometry with atmospheric pressure chemical ionization in the positive mode (UPLC–APCI⁺–MS/MS). The population analysis indicated that prostate cancer patients showed significantly higher DHEAS, DHT, E1, and E than healthy controls. Furthermore, the levels of DHEAS and E1 were significantly positively correlated with serum levels of prostate-specific antigen (PSA), which is considered to be the best conventional prostate cancer marker in the current study. In summary, we validated the use of urine samples for multiple steroid profiling by UPLC–MS/MS. Two steroids were significantly altered and correlated with PSA in the prostate cancer population. These findings open further investigation of the potential role of sex steroids in prostate cancer diagnosis. Studying large samples and determining these steroid hormones in the urine are still necessary.

Keywords:

• Urine
• prostate cancer
• sex steroids
• glucocorticoids
• ultra-high-performance liquid chromatography – tandem mass spectrometry (UPLC–MS/MS)

Ethics declarations

The authors declare that all studies conformed to ethical guidelines and were approved by the Ethics Committee of Jiangsu Provincial People’s Hospital.

Conflicts of interest

All authors declare no conflicts of interest.

Additional information

Funding

This study was supported by Natural Science Foundation of China (81872684), the Fundamental Research Funds for the Central Universities, and Funded by Southeast University "Zhongying Young Scholars" Project.