title: Simultaneous determination of cortisol, cortisone, 6β-hydroxycortisol and 6β-hydroxycortisone by HPLC.
abstract: A specific and sensitive method based on high-performance liquid chromatography with ultraviolet absorbance detection (HPLC-UV) was developed for the simultaneous determination of urinary cortisol (F), cortisone (E), 6β-hydroxycortisol (6β-OHF) and 6β-hydroxycortisone (6β-OHE) using dexamethasone as the internal standard. The method involved solid-phase extraction of the five compounds from urine using Oasis HLB Waters cartridges with an elution solvent of ethyl acetate-diethyl ether (5 mL; 4:1, v/v), followed by 1 mol/L of NaOH (1 mL) and 1.0% acetic acid (1 mL). Separation of the five analytes was achieved within 31 min by using a reversed-phase C18 analytical column (200 × 4.6 mm, 5 µm, Agilent). A UV detector operated at 245 nm was used. According to the method validation, inter-run and intra-run precision was below 9.45% and accuracy ranged from 98.16 to 115.50%. The lower limits of quantitation were 5 ng/mL for four analytes. This is the first HPLC method that can simultaneously determine F, E, 6β-OHF and 6β-OHE in human urine. The assay was applied to research the ratio of (6β-OHF + 6β-OHE)/(F + E) as a non-invasive biomarker for the metabolism of tacrolimus.
authors: Research Institute of Drug Metabolism and Pharmacokinetics, School of Pharmaceutical Sciences, Central South University, 410013 Changsha, Hunan, P.R. China.
authors: Liyun
authors: L
authors: Zheng
authors: Research Institute of Drug Metabolism and Pharmacokinetics, School of Pharmaceutical Sciences, Central South University, 410013 Changsha, Hunan, P.R. China.
authors: Xi
authors: X
authors: Luo
authors: The Third Xiangya Hospital, Central South University, 410013 Changsha, Hunan, P.R. China.
authors: Lijun
authors: L
authors: Zhu
authors: The Third Xiangya Hospital, Central South University, 410013 Changsha, Hunan, P.R. China.
authors: Wenzhao
authors: W
authors: Xie
authors: The Third Xiangya Hospital, Central South University, 410013 Changsha, Hunan, P.R. China.
authors: Shikun
authors: S
authors: Liu
authors: Research Institute of Drug Metabolism and Pharmacokinetics, School of Pharmaceutical Sciences, Central South University, 410013 Changsha, Hunan, P.R. China chengzenengcsu@yahoo.com.cn.
authors: Zeneng
authors: Z
authors: Cheng
mesh_terms: D000328:Adult; D015415:Biomarkers / Q000652:urine; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003348:Cortisone / Q000031:analogs & derivatives* / Q000737:chemistry / Q000302:isolation & purification / Q000652:urine*; D005260:Female; D006801:Humans; D006854:Hydrocortisone / Q000031:analogs & derivatives* / Q000737:chemistry / Q000302:isolation & purification / Q000652:urine*; D016014:Linear Models; D008297:Male; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D016559:Tacrolimus / Q000378:metabolism; D055815:Young Adult
chemical_list: D015415:Biomarkers; D003348:Cortisone; D006854:Hydrocortisone; D016559:Tacrolimus