Quantitative metabolic profiling of 21 endogenous corticosteroids in urine by liquid chromatography–triple quadrupole-mass spectrometry

• Published in: Analytica chimica acta 2009-01, Vol.632 (1), p.101-108
• Main Authors: Cho, Hye-Jin, Kim, Jong Dae, Lee, Won-Yong, Chung, Bong Chul, Choi, Man Ho
• Format: Article
• Language: English
• Subjects: Adrenal Cortex Hormones - chemistry; Adrenal Cortex Hormones - urine; Aged; Analytical chemistry; Calibration; Chemistry; Chromatographic methods and physical methods associated with chromatography; Chromatography, Liquid - methods; Corticosteroids; Exact sciences and technology; Humans; Liquid chromatography; Male; Molecular Structure; Other chromatographic methods; Prostate diseases; Solid Phase Extraction; Spectrometric and optical methods; Tandem mass spectrometry; Tandem Mass Spectrometry - methods
• ISSN: 0003-2670; 1873-4324
• DOI: 10.1016/j.aca.2008.10.059

Since corticosteroid metabolism may be affected by disease states, the accurate and precise measurement of endogenous corticosteroids in urine is necessary to understand their biochemical roles. An efficient quantitative profiling of 21 endogenous corticosteroids in urine has been validated by liquid chromatography–tandem mass spectrometry (LC–MS/MS). After enzymatic hydrolysis with β-glucuronidase, samples were purified using a solid-phase extraction cartridge and then separated through a sub-2 μm particle C18 column (2.1 mm × 50 mm, 1.9 μm) and quantified within 12.1 min using a triple quadrupole MS with electrospray ionization in positive ion mode. All corticosteroids resulted in the base-line separation, which is even achieved for stereo-isomers, such as α-/β-cortol, α-/β-cortolone, and allo-tetrahydrocortisol/tetrahydrocortisol. Overall recoveries ranged from 85% to 106% with limit of quantification ranged from 0.5 to 2.0 ng mL −1 for the corticosteroids examined. The precision (% CV) and accuracy (% bias) of the assay were 1.7–7.8% and 95.1–105.4%, respectively, in 0.5–200 ng mL −1 calibration ranges ( r 2 > 0.9903), for quality-control samples containing 21 endogenous corticosteroids at three different urinary concentrations. Clinical application included quantitative analysis from patients with both prostate cancer and benign prostatic hyperplasia with altered cortisol concentrations. The described LC–MS/MS method eliminates interference from other urine components, has excellent chromatographic resolution achieved by a small particle LC column with a sufficient sensitivity to allow the profiling of both gluco- and mineralo-corticosteroids at a time.