Simultaneous determination of 6beta-hydroxycortisol and cortisol in human urine and plasma by liquid chromatography with ultraviolet absorbance detection for phenotyping the CYP3A activity.
This study describes an high-performance liquid chromatographic (HPLC)-UV method for the simultaneous determination of 6beta-hydroxycortisol (6beta-OHF) and cortisol (F) in human urine and plasma. The within-day relative standard deviation of the three concentrations for both analytes was less than 6.2%. Accuracy determined at three concentrations ranged between 95 and 107%. The extraction recoveries were 64.1+/-4.3 and 88.1+/-2.4% at three concentrations for 6beta-OHF and F in urine, respectively. The extraction recoveries were 88.7+/-1.4% at three concentrations for F in plasma. This is the first HPLC method that can simultaneously determine 6beta-OHF and F in human urine and plasma and is suitable for routine assessment of the CYP3A activity expressed as 6beta-hydroxylation clearance.
Hu
Zheyi
Z
Department of Biopharmaceutics, School of Pharmaceutical Sciences, Central South University, Changsha, Hunan 410013, PR China.
Gong
Qian
Q
Hu
Xiaolei
X
Wang
Liqing
L
Cao
Yajie
Y
Cao
Wei
W
Yu
Qi
Q
Cheng
Zeneng
Z
D000328:Adult; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002940:Circadian Rhythm; D051544:Cytochrome P-450 CYP3A / Q000235:genetics*; D005260:Female; D006801:Humans; D006854:Hydrocortisone / Q000031:analogs & derivatives* / Q000032:analysis* / Q000097:blood / Q000652:urine; D008297:Male; D010641:Phenotype; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013056:Spectrophotometry, Ultraviolet
C001380:6 beta-hydroxycortisol; D051544:Cytochrome P-450 CYP3A; D006854:Hydrocortisone