Parallel derivatization strategy coupled with liquid chromatography-mass spectrometry for broad coverage of steroid hormones

J Chromatogr A. 2020 Mar 15:1614:460709. doi: 10.1016/j.chroma.2019.460709. Epub 2019 Nov 13.

Abstract

Steroid hormones are a type of crucial substances that mediate numerous vital physiological functions. The comprehensive detection of steroid hormones can help understand the physiopathologic mechanism of steroid hormone-related diseases. It is very difficult to determine steroid hormones in biological samples due to their low endogenous concentrations and poor ionization efficiency. In this study, an efficient and sensitive approach was developed for profiling steroid hormones by combining liquid-liquid extraction and parallel derivatization with liquid chromatography-tandem mass spectrometry. Methoxyamine and dansyl chloride were used to derivatize steroid hormones containing carbonyl and phenolic hydroxyl groups, respectively. Our established method achieved simultaneous analysis of carbonyl and phenolic hydroxyl-containing steroid hormones and could cover estrogens, androgens, corticoids and progestogens. Twenty-nine steroid hormones were detected at pg/mL levels with the sensitivity enhanced by three orders of magnitude after derivatization. The linearity (with linear range of 2-4 orders of magnitude), precision (less than 15%) and recovery (71.1-128.7%) were satisfactory for quantitative analysis of steroid hormones. Finally, the established method was successfully employed to the determination of steroid hormones in serum samples of healthy males and females as well as ovarian cancer patients. The results showed that this approach was suitable and reliable for routine test of steroid hormones containing carbonyl and phenolic hydroxyl groups.

Keywords: Liquid chromatography-mass spectrometry; Parallel derivatization; Simultaneous quantification; Steroid hormones.

MeSH terms

  • Chromatography, High Pressure Liquid / methods*
  • Dansyl Compounds / chemistry
  • Female
  • Humans
  • Liquid-Liquid Extraction
  • Male
  • Mass Spectrometry / methods*
  • Methoxamine / chemistry
  • Ovarian Neoplasms / blood
  • Ovarian Neoplasms / pathology
  • Progestins / blood
  • Progestins / chemistry
  • Progestins / isolation & purification
  • Steroids / blood
  • Steroids / chemistry*
  • Steroids / isolation & purification

Substances

  • Dansyl Compounds
  • Progestins
  • Steroids
  • Methoxamine
  • dansyl chloride