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

Hye Jin Cho, Jong Dae Kim, Won Yong Lee, Bong Chul Chung, Man Ho Choi

Research output: Contribution to journalArticle

68 Citations (Scopus)

Abstract

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 (r2 > 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.

Original languageEnglish
Pages (from-to)101-108
Number of pages8
JournalAnalytica Chimica Acta
Volume632
Issue number1
DOIs
Publication statusPublished - 2009 Jan 19

Fingerprint

Liquid chromatography
Liquid Chromatography
urine
Mass spectrometry
liquid chromatography
Mass Spectrometry
Adrenal Cortex Hormones
mass spectrometry
Urine
Tetrahydrocortisol
quality control
quantitative analysis
hydrolysis
cancer
ionization
metabolism
assay
calibration
ion
Electrospray ionization

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry
  • Biochemistry
  • Environmental Chemistry
  • Spectroscopy

Cite this

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abstract = "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 (r2 > 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.",
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Quantitative metabolic profiling of 21 endogenous corticosteroids in urine by liquid chromatography-triple quadrupole-mass spectrometry. / Cho, Hye Jin; Kim, Jong Dae; Lee, Won Yong; Chung, Bong Chul; Choi, Man Ho.

In: Analytica Chimica Acta, Vol. 632, No. 1, 19.01.2009, p. 101-108.

Research output: Contribution to journalArticle

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