2. Academic Validation
  3. Development and validation of a novel LC-MS/MS method for simultaneous determination of abiraterone and its seven steroidal metabolites in human serum: Innovation in separation of diastereoisomers without use of a chiral column

Development and validation of a novel LC-MS/MS method for simultaneous determination of abiraterone and its seven steroidal metabolites in human serum: Innovation in separation of diastereoisomers without use of a chiral column

  • J Steroid Biochem Mol Biol. 2017 Sep;172:231-239. doi: 10.1016/j.jsbmb.2016.04.002.
Mohammad Alyamani 1,  Zhenfei Li 2,  Sunil K Upadhyay 3,  David J Anderson 4,  Richard J Auchus 3,  Nima Sharifi 5
Affiliations

Affiliations

  • 1 Department of Cancer Biology, Lerner Research Institute, Cleveland Clinic, Cleveland, OH, United States; Department of Chemistry, Cleveland State University, Cleveland, OH, United States.
  • 2 Department of Cancer Biology, Lerner Research Institute, Cleveland Clinic, Cleveland, OH, United States.
  • 3 Division of Endocrinology and Metabolism, Department of Internal Medicine and Department of Pharmacology, University of Michigan Medical School, Ann Arbor, MI, United States.
  • 4 Department of Chemistry, Cleveland State University, Cleveland, OH, United States.
  • 5 Department of Cancer Biology, Lerner Research Institute, Cleveland Clinic, Cleveland, OH, United States; Department of Urology, Glickman Urological and Kidney Institute, Cleveland Clinic, Cleveland, OH, United States; Department of Hematology and Oncology, Taussig Cancer Institute, Cleveland Clinic, Cleveland, OH, United States; Department of Chemistry, Cleveland State University, Cleveland, OH, United States. Electronic address: sharifn@ccf.org.
PMID: 27063554 DOI: 10.1016/j.jsbmb.2016.04.002
Abstract

Abiraterone acetate (AA), the prodrug of abiraterone, is FDA-approved for the treatment of castration-resistant prostate Cancer. Abiraterone is metabolized in patients to a more potent analogue, D4A. However, we have recently reported that this analogue is further metabolized to additional metabolites in patients treated with AA. Here, we present a liquid chromatography-tandem mass spectrometry method developed to resolve and detect abiraterone and its seven metabolites in human serum using an AB Sciex Qtrap 5500 mass analyzer coupled with a Shimadzu Nexera UPLC station. Analytes and the internal standard (abiraterone-d4) were extracted from human serum using the liquid-liquid extraction procedure. The analytes were separated using a Zorbax Eclipse Plus C18 150×2.1mm, 3.5μm column at 40°C and an isocratic mobile phase 35% A (0.1% formic acid in water), 65% B (0.1% formic acid in methanol:acetonitrile; 60:40). Electrospray ionization in positive mode was applied with multiple reaction monitoring in a total run time of 13min. Abiraterone detection was linear in the range 2-400ng/mL and all metabolites from 0.1-20ng/mL. The method was validated following US FDA guidelines for bioanalytical method validation, and all the metabolite results were within the acceptance limits. Despite the similarity in structure and mass transition between the metabolites, the validated method separated all the metabolites, including diastereomers, to allow accurate identification and quantitation of each compound.

Keywords

Abiraterone; LC–MS/MS; Metabolites; Method validation; Prostate cancer.

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