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

doi:10.1016/j.jsbmb.2016.04.002

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

Research output: Contribution to journal › Article › peer-review

Abstract

<p> <p id="x-x-x-x-spar0075"> 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 <a href="https://proxy.ulib.csuohio.edu:2062/topics/biochemistry-genetics-and-molecular-biology/liquid-chromatography-tandem-mass-spectrometry" title="Learn more about Liquid Chromatography Tandem Mass Spectrometry from ScienceDirect's AI-generated Topic Pages"> liquid chromatography-tandem mass spectrometry </a> 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 <a href="https://proxy.ulib.csuohio.edu:2062/topics/biochemistry-genetics-and-molecular-biology/liquid-liquid-extraction" title="Learn more about Liquid Liquid Extraction from ScienceDirect's AI-generated Topic Pages"> liquid–liquid extraction </a> 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 <a href="https://proxy.ulib.csuohio.edu:2062/topics/biochemistry-genetics-and-molecular-biology/mobile-phase-composition" title="Learn more about Mobile Phase Composition from ScienceDirect's AI-generated Topic Pages"> mobile phase </a> 35% A (0.1% formic acid in water), 65% B (0.1% formic acid in methanol:acetonitrile; 60:40). <a href="https://proxy.ulib.csuohio.edu:2062/topics/biochemistry-genetics-and-molecular-biology/electrospray-ionization" title="Learn more about Electrospray Ionization from ScienceDirect's AI-generated Topic Pages"> Electrospray ionization </a> 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 <a href="https://proxy.ulib.csuohio.edu:2062/topics/biochemistry-genetics-and-molecular-biology/diastereoisomer" title="Learn more about Diastereoisomer from ScienceDirect's AI-generated Topic Pages"> diastereomers </a> , to allow accurate identification and quantitation of each compound. </p></p>

Original language	American English
Journal	Journal of Steroid Biochemistry and Molecular Biology
Volume	172
DOIs	https://doi.org/10.1016/j.jsbmb.2016.04.002
State	Published - Sep 1 2017

Keywords

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

Disciplines

Analytical Chemistry
Biochemistry

Access to Document

10.1016/j.jsbmb.2016.04.002License: CC BY

Development and Validation of a Novel LC MS MS Method for SimultaFinal published version, 2.2 MBLicense: CC BY

Cite this

Alyamani, M., Li, Z., Upadhyay, S. K., Anderson, D. J., Auchus, R. J., & Sharifi, N. (2017). 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. Journal of Steroid Biochemistry and Molecular Biology, 172. https://doi.org/10.1016/j.jsbmb.2016.04.002

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. / Alyamani, Mohammad; Li, Zhenfei; Upadhyay, Sunil K. et al.
In: Journal of Steroid Biochemistry and Molecular Biology, Vol. 172, 01.09.2017.

Research output: Contribution to journal › Article › peer-review

Alyamani, M, Li, Z, Upadhyay, SK, Anderson, DJ, Auchus, RJ & Sharifi, N 2017, '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', Journal of Steroid Biochemistry and Molecular Biology, vol. 172. https://doi.org/10.1016/j.jsbmb.2016.04.002

Alyamani M, Li Z, Upadhyay SK, Anderson DJ, Auchus RJ, Sharifi N. 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. Journal of Steroid Biochemistry and Molecular Biology. 2017 Sep 1;172. doi: 10.1016/j.jsbmb.2016.04.002

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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 = "Prostate cancer; Abiraterone; LC–MS/MS; Method validation;Metabolites",

author = "Mohammad Alyamani and Zhenfei Li and Upadhyay, \{Sunil K.\} and Anderson, \{David J.\} and Auchus, \{Richard J.\} and Nima Sharifi",

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AU - Alyamani, Mohammad

AU - Li, Zhenfei

AU - Upadhyay, Sunil K.

AU - Anderson, David J.

AU - Auchus, Richard J.

AU - Sharifi, Nima

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

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JO - Journal of Steroid Biochemistry and Molecular Biology

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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

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