Simultaneous Determination of 6β-Hydroxycortisol and 6β-Hydroxycortisone in Human Urine by LC with UV Absorbance Detection

Luo, Xi; Zhu, Lijun; Wu, Wei; Sheng, Xia; Cai, Ningfang; Liu, Shikun; Zhang, Cheng

doi:10.1365/s10337-009-1267-9

Cited by 3 publications

(3 citation statements)

References 21 publications

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“…6β‐Hydroxycortisol, 6β‐hydroxycortisone, F, and E in urine were measured using the HPLC–UV method developed in our laboratory . The lower limit of quantitation (LLOQ) was 5 μg L −1 for all compounds.…”

Section: Methodsmentioning

confidence: 99%

Evaluation of 6β-Hydroxycortisol and 6β-Hydroxycortisone as Biomarkers for Cytochrome P450 3A Activity: Insight into Their Predictive Value for Estimating Oral Immunosuppressant Metabolism

Luo

Zheng

Cai

et al. 2015

Journal of Pharmaceutical Sciences

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Section: Methodsmentioning

confidence: 99%

Evaluation of 6β-Hydroxycortisol and 6β-Hydroxycortisone as Biomarkers for Cytochrome P450 3A Activity: Insight into Their Predictive Value for Estimating Oral Immunosuppressant Metabolism

Luo

Zheng

Cai

et al. 2015

Journal of Pharmaceutical Sciences

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“…1 Chromatograms of a standard glucocorticoids with 10 lg mL -1 6b-OHF, 10 lg mL -1 E and 2 lg mL -1 F, analyzed by Acquity UPLC BEH phenyl column; b standard glucocorticoids with 10 lg mL -1 6b-OHF, 10 lg mL -1 E and 2 lg mL -1 F, analyzed by Acquity UPLC BEH C18 column; c spot urine from a healthy volunteer spiked with 300 ng mL -1 6b-OHF, 300 ng mL -1 E and 75 ng mL -1 F; d spot urine from a healthy volunteer. Peaks: 1 6b-OHF, 2 E, 3 F, 4 dexamethasone (IS) consequences and using a spot urine sample may be convenient approach [11].…”

Section: Clinical Applicationmentioning

confidence: 99%

“…Recently, Luo et al [11] have reported that the ratio of 6b-OHF?6b-hydrocortisone (6b-OHE)/F?E may be a a Values represent the mean ± SD b Accuracy (%) = (mean concentration measured in spiked urine/standard concentration spiked into urine) 9 100% c RSD, relative standard deviation (%) = (SD/mean) 9 100 Values represent the mean ± SD * P \ 0.01; ** P \ 0.001 compared with baseline by Bonferroni tests better index for estimating the variability in CYP3A activity, however, the determination required two separate LC-UV methods. 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.…”

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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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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Simultaneous Determination of Cortisol, Cortisone, 6 -Hydroxycortisol and 6 -Hydroxycortisone by HPLC

Zheng

Luo

Zhu

et al. 2015

Journal of Chromatographic Science

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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.

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Simultaneous Determination of 6β-Hydroxycortisol and 6β-Hydroxycortisone in Human Urine by LC with UV Absorbance Detection

Cited by 3 publications

References 21 publications

Evaluation of 6β-Hydroxycortisol and 6β-Hydroxycortisone as Biomarkers for Cytochrome P450 3A Activity: Insight into Their Predictive Value for Estimating Oral Immunosuppressant Metabolism

Evaluation of 6β-Hydroxycortisol and 6β-Hydroxycortisone as Biomarkers for Cytochrome P450 3A Activity: Insight into Their Predictive Value for Estimating Oral Immunosuppressant Metabolism

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, 6 -Hydroxycortisol and 6 -Hydroxycortisone by HPLC

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