Simultaneous determination of 6β-hydroxycortisol and cortisol in human urine and plasma by liquid chromatography with ultraviolet absorbance detection for phenotyping the CYP3A activity

Hu, Zheyi; Gong, Qian; Hu, Xiaolei; Wang, Liqing; Cao, Yajie; Cao, Wei; Yu, Qi; Zhang, Cheng

doi:10.1016/j.jchromb.2005.09.007

Cited by 25 publications

(24 citation statements)

References 18 publications

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“…The urine F concentration decreased during the afternoon and the urinary 6b-OHF/F ratio exhibited significant diurnal variation with a peak between 1,300 and 2,100 (Table 3). These results are in good agreement with previous reports [12,13]. It was suggested that the diurnal variation of the urinary 6b-OHF/F ratio may infer the circadian rhythm of hepatic CYP3A activity [12] although it may just reflect the variation in the availability of the substrate F.…”

Section: Clinical Applicationsupporting

confidence: 92%

“…Here we describe a simple, cost-effective and rapid method for simultaneous determination of F, E and 6b-OHF in human urine with adequate sensitivity using UPLC-UV detection. By taking advantage of UPLC techniques on smaller particle size (1.7 lm) and higher operating pressures, this method can offer efficient chromatography for the simultaneous determination of F, E, and 6b-OHF in human urine with reduced run time (7 min) compared to traditional HPLC method with a long analysis time of about 27 min [8,13].…”

Section: Clinical Applicationmentioning

confidence: 99%

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Simultaneous Determination of Cortisol, Cortisone, and 6β-Hydroxycortisol in Human Urine by UPLC with UV Detector and Application to Determine Diurnal Variations

Xiao

Chan

et al. 2011

Chromatographia

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In the present studies, a simple rapid ultra performance liquid chromatography (UPLC) method with UV detection for the simultaneous determination of cortisol, cortisone and 6b-hydroxycortisol in human urine was developed. The three analytes and the internal standard dexamethasone were separated on a Waters Acquity UPLC-Tunable UV system with an Acquity UPLC BEH C18 column (50 9 2.1 mm ID, 1.7 lm) using a gradient elution of methanol and water (containing 0.01% formic acid) with a run time of 7 min. The method was accurate and precise, over the ranges of 5-200 ng mL -1 for cortisol, and 10-1,000 ng mL -1 for both cortisone and 6b-hydroxycortisol, and showed good linearity (r 2 [ 0.999). This method was applied for the measurement of cortisol, cortisone and 6b-hydroxycortisol in samples collected over different periods as a tool to assess the activity of CYP3A and 11b-hydroxysteroid dehydrogenase type 2 enzymes.

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Section: Clinical Applicationsupporting

confidence: 92%

Section: Clinical Applicationmentioning

confidence: 99%

Simultaneous Determination of Cortisol, Cortisone, and 6β-Hydroxycortisol in Human Urine by UPLC with UV Detector and Application to Determine Diurnal Variations

Xiao

Chan

et al. 2011

Chromatographia

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“…Blood and urine samples were collected in the same way. Analytical methods 6β-OHF and F in plasma or urine F and 6β-OHF in urine or plasma were measured using the HPLC-UV method that was developed in our laboratory [12] . The lower limit of quantitation was 6.9 μg/L for 6β-OHF and 2 μg/L for F in urine.…”

Section: Experimental Protocolmentioning

confidence: 99%

“…We have previously published the results obtained using a high-performance liquid chromatography with ultraviolet absorbance detection (HPLC-UV) method for the simultaneous determination of 6β-OHF and F in human urine or plasma [12] . Of the 12 subjects (6 males and 6 females) enrolled in our study, a good correlation between cortisol 6β-hydroxylation clearance and MDZ oral clearance was seen in only 6 male subjects.…”

Section: Introductionmentioning

confidence: 99%

Evaluation of CYP3A activity in humans using three different parameters based on endogenous cortisol metabolism

Luo

et al. 2009

Acta Pharmacol Sin

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Aim: Currently, there is considerable debate as to which method is more accurate for measuring the activity of CYP3A in vivo: cortisol 6β-hydroxylation clearance (Cl m(6β) ) or the urinary ratio of 6β-OHF to F (6β-OHF/F). Furthermore, the value of measuring endogenous levels of cortisol over a 24 h period (AUC F ) needs to be confirmed. The aim of the present study was to determine which method was most effective at measuring changes in the in vivo activity of CYP3A: AUC F , Cl m(6β) , or 6β-OHF/F. Methods: A two phase, cross-over design was adopted in this study. A total of 24 subjects (12 males and 12 females) were randomly assigned to one of two groups: the test group subjects were given 250 mg clarithromycin tablets twice a day for a period of 4 d, whereas the control group received a placebo twice daily for a similar period. On d 5 of the study, the last dose of either clarithromycin or placebo was supplemented with an oral dose of 7.5 mg midazolam (MDZ); blood and urine samples were then collected at various times. All samples collected at the same sampling times on d 4 were used to evaluate the effects of MDZ administration on cortisol levels and metabolism. The ratio of 1-hydroxymidazolam (1-OHMDZ) concentration to MDZ concentration at 1 h (MR) was taken as a measure of the in vivo CYP3A activity. AUC F , Cl m(6β) , and 6β-OHF/F were also used as biomarkers for CYP3A activity. Results: No correlations were found (either before or after inhibition) between CYP3A activity and any of the following measures: AUC F , Cl m(6β) , or 6β-OHF/F (r<0.4, P>0.05). After 4 d of clarithromycin administration, CYP3A activity (MR) decreased by 75% (P=0.000), whereas AUC F increased by 19% (P=0.040), and Cl m(6β) and 6β-OHF/F decreased by 54.2% (P=0.000) and 50% (P=0.003), respectively. No significant changes in AUC F (P=0.178), or in the amount of urinary 6β-OHF (P=0.169), or in F (P=0.391) were found over a 24 h time period, either with or without MDZ administration. Conclusion: Although Cl m(6β) and 6β-OHF/F can reflect the decline in CYP3A activity, the impression they provide is neither accurate nor complete. AUC F is completely ineffective for evaluating variations in CYP3A activity. MDZ administration had no evident effects on either cortisol metabolism or excretion over a period of 24 h.

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“…Two-step reaction to produce bismethylenedioxy-3,11-dipentafluoropropionyl derivatives was suggested to avoid the problem with stability and to ensure better resolution [28]. HPLC-UV with normal phase [29] and reversed phase [16,[30][31][32][33][34] was applied to determine glucocorticoids in clinical samples. LC-MS-MS methods were presented for simultaneous determination of THF, allo-THF and THE in urine with the lowest concentration detected ranging from 0.1 ng mL −1 [12] to 0.5 ng mL −1 [35].…”

Section: Introductionmentioning

confidence: 99%

HPLC method for determination of fluorescence derivatives of cortisol, cortisone and their tetrahydro- and allo-tetrahydro-metabolites in biological fluids

Główka

Kosicka

Karaźniewicz−Łada

2010

Journal of Chromatography B

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Simultaneous determination of 6β-hydroxycortisol and cortisol in human urine and plasma by liquid chromatography with ultraviolet absorbance detection for phenotyping the CYP3A activity

Cited by 25 publications

References 18 publications

Simultaneous Determination of Cortisol, Cortisone, and 6β-Hydroxycortisol in Human Urine by UPLC with UV Detector and Application to Determine Diurnal Variations

Simultaneous Determination of Cortisol, Cortisone, and 6β-Hydroxycortisol in Human Urine by UPLC with UV Detector and Application to Determine Diurnal Variations

Evaluation of CYP3A activity in humans using three different parameters based on endogenous cortisol metabolism

HPLC method for determination of fluorescence derivatives of cortisol, cortisone and their tetrahydro- and allo-tetrahydro-metabolites in biological fluids

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