Simultaneous determination of tetrahydrocortisol, allotetrahydrocortisol and tetrahydrocortisone in human urine by liquid chromatography-electrospray ionization tandem mass spectrometry

• Published in: Steroids Vol. 73; no. 7; pp. 727 - 737
• Main Authors: Yamashita, Kouwa, Nakagawa, Risa, Okuyama, Mitsunobu, Honma, Seijiro, Takahashi, Madoka, Numazawa, Mitsuteru
• Format: Journal Article
• Language: English
• Published: NEW YORK Elsevier Inc 01.08.2008; Elsevier; Elsevier Science
• Subjects: Biochemistry & Molecular Biology; Biological and medical sciences; Calibration; Chromatography, Liquid - methods; Electrospray ionization; Endocrinology & Metabolism; Fundamental and applied biological sciences. Psychology; Humans; Life Sciences & Biomedicine; Liquid chromatography-tandem mass spectrometry; Picolinyl derivative; Reproducibility of Results; Science & Technology; Sensitivity and Specificity; Spectrometry, Mass, Electrospray Ionization - methods; Tandem Mass Spectrometry; Tetrahydrocortisol - analogs & derivatives; Tetrahydrocortisol - chemistry; Tetrahydrocortisol - urine; Tetrahydrocortisone - chemistry; Tetrahydrocortisone - urine; Urinary corticosteroid metabolite; Vertebrates: endocrinology; Liquid chromatography-tandem mass spectrometry; Electrospray ionization; Picolinyl derivative; Urinary corticosteroid metabolite; ESTROGENS; STEROIDS; QUANTIFICATION; BETA-HYDROXYSTEROID DEHYDROGENASE; electrospray ionization; DERIVATIZATION; picolinyl derivative; 11-BETA-HYDROXYSTEROID DEHYDROGENASE; liquid chromatography-tandem mass spectrometry; urinary corticosteroid metabolite; CORTICOSTEROIDS; DERIVATIVES; PRE-RECEPTOR REGULATION; SOLID-PHASE EXTRACTION; Human; Urine; Biological fluid; Corticosteroid; Metabolite; Liquid chromatography-tandem mass spectrometry,Electrospray ionization,Picolinyl derivative,Urinary corticosteroid metabolite; Mass spectrometry
• ISSN: 0039-128X; 1878-5867
• DOI: 10.1016/j.steroids.2008.02.009

Simultaneous quantification method of three major metabolites of cortisone and cortisol, tetrahydrocortisol, allotetrahydrocortisol and tetrahydrocortisone by liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) was investigated in a positive mode using a recently developed picolinyl derivatization. Conversion of each steroid into the corresponding picolinyl derivatives ( 1b, 2b or 3b) was performed by mixed anhydride method using picolinic acids and 2-methyl-6-nitrobenzoic anhydride. Derivatization proceeded smoothly to afford the corresponding 3, 21-dipicolinyl derivatives. Positive ion-ESI mass spectra of the picolinyl derivatives were dominated by an appearance of [M + H] + as base peaks in all cases. The picolinyl derivatives provided 15 to 80-fold higher ESI response in the LC-ESI-MS/MS (selected reaction monitoring: SRM) when compared to those of underivatized molecules in a positive LC-ESI mode. The use of the picolinyl ester, solid-phase extraction, and deuterium labeled internal standards enabled the concentrations of these metabolites in human urine to be determined simultaneously by LC-ESI-MS/MS (SRM) with a small sample volume of less than 1 μl urine.