Simultaneous determination of 6beta- and 6alpha-hydroxycortisols and 6beta-hydroxycortisone in human urine by stable isotope dilution mass spectrometry.
A capillary gas chromatographic-mass spectrometric method for the simultaneous determination of 6beta-hydroxycortisol (6beta-OHF, 6beta,11beta,17alpha,21-tetrahydroxypregn-4-ene- 3,20-dione), 6alpha-hydroxycortisol (6alpha-OHF, 6alpha,11beta,17alpha,21-tetrahydroxypregn-4-ene -3,20-dione) and 6beta-hydroxycortisone (6beta-OHE, 6beta,17alpha,21-trihydroxypregn-4-ene-3,11,20-trione) in human urine is described. Deuterium-labelled compounds, 6beta-[1,1,19,19,19-2H5]OHF (6beta-OHF-d5), 6alpha-[1,1,19,19,19-2H5]OHF (6alpha-OHF-d5) and 6beta-[1,1,19,19,19-2H5]OHE (6beta-OHE-d5) were used as internal standards. Quantitation was carried out by selected-ion monitoring of the characteristic fragment ions ([M-31]+) of the methoxime-trimethylsilyl (MO-TMS) derivatives of 6beta-OHF, 6alpha-OHF and 6beta-OHE. The sensitivity, specificity, precision and accuracy of the method were demonstrated to be satisfactory for measuring 6beta-OHF, 6alpha-OHF and 6beta-OHE in human urine.
Furuta
T
T
Department of Medicinal Chemistry and Clinical Pharmacy, School of Pharmacy, Tokyo University of Pharmacy and Life Science, Japan. furutat@ps.toyaku.ac.jp
Matsuzawa
M
M
Shibasaki
H
H
Kasuya
Y
Y
D002138:Calibration; D006801:Humans; D006854:Hydrocortisone / Q000031:analogs & derivatives / Q000652:urine*; D007554:Isotopes; D013058:Mass Spectrometry / Q000379:methods*; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D007554:Isotopes; D006854:Hydrocortisone