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

Mohammad Alyamani, Zhenfei Li, Sunil K. Upadhyay, David J. Anderson, Richard J. Auchus, Nima Sharifi

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12 Citations (Scopus)

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. ×. 2.1. mm, 3.5. μm 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 13. min. Abiraterone detection was linear in the range 2-400. ng/mL and all metabolites from 0.1-20. ng/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.

Original languageEnglish (US)
JournalJournal of Steroid Biochemistry and Molecular Biology
DOIs
StateAccepted/In press - Feb 24 2016

Fingerprint

Metabolites
formic acid
Innovation
Serum
Liquid-Liquid Extraction
Castration
Prodrugs
Electrospray ionization
Tandem Mass Spectrometry
Liquid Chromatography
Liquid chromatography
Liquids
Methanol
Prostatic Neoplasms
Mass spectrometry
abiraterone
Guidelines
Water
Monitoring
Abiraterone Acetate

Keywords

  • Abiraterone
  • LC-MS/MS
  • Metabolites
  • Method validation
  • Prostate cancer

ASJC Scopus subject areas

  • Biochemistry
  • Clinical Biochemistry
  • Endocrinology
  • Cell Biology
  • Molecular Biology
  • Endocrinology, Diabetes and Metabolism
  • Molecular Medicine

Cite this

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title = "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",
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. ×. 2.1. mm, 3.5. μm 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 13. min. Abiraterone detection was linear in the range 2-400. ng/mL and all metabolites from 0.1-20. ng/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",
author = "Mohammad Alyamani and Zhenfei Li and Upadhyay, {Sunil K.} and Anderson, {David J.} and Auchus, {Richard J.} and Nima Sharifi",
year = "2016",
month = "2",
day = "24",
doi = "10.1016/j.jsbmb.2016.04.002",
language = "English (US)",
journal = "Journal of Steroid Biochemistry and Molecular Biology",
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T1 - Development and validation of a novel LC-MS/MS method for simultaneous determination of abiraterone and its seven steroidal metabolites in human serum

T2 - Innovation in separation of diastereoisomers without use of a chiral column

AU - Alyamani, Mohammad

AU - Li, Zhenfei

AU - Upadhyay, Sunil K.

AU - Anderson, David J.

AU - Auchus, Richard J.

AU - Sharifi, Nima

PY - 2016/2/24

Y1 - 2016/2/24

N2 - 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. ×. 2.1. mm, 3.5. μm 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 13. min. Abiraterone detection was linear in the range 2-400. ng/mL and all metabolites from 0.1-20. ng/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.

AB - 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. ×. 2.1. mm, 3.5. μm 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 13. min. Abiraterone detection was linear in the range 2-400. ng/mL and all metabolites from 0.1-20. ng/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.

KW - Abiraterone

KW - LC-MS/MS

KW - Metabolites

KW - Method validation

KW - Prostate cancer

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