title: UHPLC-MS/MS bioanalysis of urinary DHEA, cortisone and their hydroxylated metabolites as potential biomarkers for CYP3A-mediated drug-drug interactions.
abstract: OBJECTIVE
  A UHPLC-MS/MS assay was developed to quantify urinary dehydroepiandrosterone (DHEA), 7β-hydroxy-DHEA, cortisone and 6β-hydroxycortisone as potential biomarkers to predict CYP3A activity.
  
  
  RESULTS
  A sensitive assay at LLOQ of 0.500 ng/ml with good accuracy and precision was developed for the four analytes in human urine. This UHPLC-MS/MS assay was optimized by eliminating nonspecific loss of the analytes in urine, ensuring complete hydrolysis of the conjugates to unconjugated forms and use of the product ions of [M+H-H2O]+for multiple reaction monitoring detection of DHEA and 7β-hydroxy-DHEA.
  
  
  CONCLUSIONS
  This assay was successfully applied to a pilot clinical study. It is also suitable for future drug-drug interaction studies to continue evaluating the potential of these steroids as biomarkers for CYP3A inhibition and induction.
authors.affiliation: Analytical & Bioanalytical Operations, Bristol-Myers Squibb Company, Princeton, NJ 08543, USA.
authors.forename: Naiyu
authors.initials: N
authors.lastname: Zheng
authors.affiliation: Pharmaceutical Candidate Optimization, Bristol-Myers Squibb Company, Princeton, NJ 08543, USA.
authors.forename: Lisa J
authors.initials: LJ
authors.lastname: Christopher
authors.affiliation: Early Clinical & Translational Research, Bristol-Myers Squibb Company, Princeton, NJ 08543, USA.
authors.forename: Xuewen
authors.initials: X
authors.lastname: Ma
authors.affiliation: Analytical & Bioanalytical Operations, Bristol-Myers Squibb Company, Princeton, NJ 08543, USA.
authors.forename: Qin C
authors.initials: QC
authors.lastname: Ji
authors.affiliation: Analytical & Bioanalytical Operations, Bristol-Myers Squibb Company, Princeton, NJ 08543, USA.
authors.forename: Adela
authors.initials: A
authors.lastname: Buzescu
authors.affiliation: Analytical & Bioanalytical Operations, Bristol-Myers Squibb Company, Princeton, NJ 08543, USA.
authors.forename: Hamza
authors.initials: H
authors.lastname: Kandoussi
authors.affiliation: Early Clinical & Translational Research, Bristol-Myers Squibb Company, Princeton, NJ 08543, USA.
authors.forename: Samira M
authors.initials: SM
authors.lastname: Garonzik
authors.affiliation: Early Clinical & Translational Research, Bristol-Myers Squibb Company, Princeton, NJ 08543, USA.
authors.forename: Frank
authors.initials: F
authors.lastname: LaCreta
authors.affiliation: Analytical & Bioanalytical Operations, Bristol-Myers Squibb Company, Princeton, NJ 08543, USA.
authors.forename: Anne-Francoise
authors.initials: AF
authors.lastname: Aubry
authors.affiliation: Analytical & Bioanalytical Operations, Bristol-Myers Squibb Company, Princeton, NJ 08543, USA.
authors.forename: Mark E
authors.initials: ME
authors.lastname: Arnold
authors.affiliation: Analytical & Bioanalytical Operations, Bristol-Myers Squibb Company, Princeton, NJ 08543, USA.
authors.forename: Jianing
authors.initials: J
authors.lastname: Zeng
mesh_terms: D015415:Biomarkers / Q000652:urine*; D002851:Chromatography, High Pressure Liquid / Q000592:standards; D003348:Cortisone / Q000378:metabolism / Q000592:standards / Q000652:urine*; D051544:Cytochrome P-450 CYP3A / Q000737:chemistry / Q000378:metabolism*; D003687:Dehydroepiandrosterone / Q000378:metabolism / Q000592:standards / Q000652:urine*; D004347:Drug Interactions; D006801:Humans; D006900:Hydroxylation; D057230:Limit of Detection; D059625:Liquid-Liquid Extraction; D011786:Quality Control; D053719:Tandem Mass Spectrometry / Q000592:standards; D016482:Urinalysis / Q000295:instrumentation / Q000379:methods*
chemical_list: D015415:Biomarkers; D003687:Dehydroepiandrosterone; D051544:Cytochrome P-450 CYP3A; D003348:Cortisone
keywords: 6β-OH-cortisone; 7β-OH-DHEA; CYP3A induction and inhibition; DHEA; cortisone; drug–drug interactions (DDIs); human urine