A validated enantioselective LC-MS/MS assay for quantification of a major chiral metabolite of an achiral 11-β-hydroxysteroid-dehydrogenase 1 inhibitor in human plasma

Application to a clinical pharmacokinetic study

Bruce ARNOLD, Wendy BONYTHON

Research output: A Conference proceeding or a Chapter in BookChapter

3 Citations (Scopus)

Abstract

BMS-823778 is a potent 11-β-hydroxysteroid-dehydrogenase 1 (11βHSD-1) inhibitor and a potential therapeutic agent for type 2 diabetes mellitus (T2DM). A high performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay was developed and validated to enable reliable separation and quantification of both enantiomers of a chiral hydroxy metabolite (BMT-094817) in human plasma. Following liquid-liquid extraction in a 96-well plate format, chromatographic separation of the metabolite enantiomers was achieved by isocratic elution on a Chiralpak IA-3 column. Chromatographic conditions were optimized to ensure separation of both metabolite enantiomers. Metabolite enantiomers and stable isotope-labeled (SIL) internal standards were detected by positive ion electrospray tandem mass spectrometry. The LC-MS/MS assay was validated over a concentration range of 0.200-200 ng/mL. Intra- and inter-assay precision values for replicate quality control samples were less than 9.9% for both enantiomers during the assay validation. Mean quality control accuracy values were within ±7.3%. Assay recoveries were high (>75%) and consistent across the assay range. The metabolite enantiomers were stable in human blood for 2 h on ice. The analytes were also stable in human plasma for 25 h at room temperature, 34 days at -20 °C and -70 °C, and following five freeze-thaw cycles. No interconversion of the metabolite enantiomers was detected under any bioanalytical stress conditions, from blood collection/processing through extracted sample storage. The validated assay was successfully applied to the quantification of both metabolite enantiomers in human plasma in support of a human pharmacokinetic study.

Original languageEnglish
Title of host publicationFinancial Crimes: Psychological, Technological, and Ethical Issues
EditorsMichel Dion, David Weisstub, Jean-Loup Richet
Place of PublicationSwitzerland
PublisherSpringer
Pages167-172
Number of pages6
Volume1022
ISBN (Print)9783319324180
DOIs
Publication statusPublished - 2016

Publication series

NameJournal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
ISSN (Print)1570-0232

Fingerprint

11-beta-Hydroxysteroid Dehydrogenases
Plasma applications
Pharmacokinetics
Enantiomers
Metabolites
Assays
Plasma (human)
Tandem Mass Spectrometry
Quality Control
Liquid-Liquid Extraction
Quality control
Mass spectrometry
Ice
Blood
Isotopes
Type 2 Diabetes Mellitus
High Pressure Liquid Chromatography
Clinical Studies
Ions
High performance liquid chromatography

Cite this

ARNOLD, B., & BONYTHON, W. (2016). A validated enantioselective LC-MS/MS assay for quantification of a major chiral metabolite of an achiral 11-β-hydroxysteroid-dehydrogenase 1 inhibitor in human plasma: Application to a clinical pharmacokinetic study. In M. Dion, D. Weisstub, & J-L. Richet (Eds.), Financial Crimes: Psychological, Technological, and Ethical Issues (Vol. 1022, pp. 167-172). (Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences). Switzerland: Springer. https://doi.org/10.1016/j.jchromb.2016.03.043
ARNOLD, Bruce ; BONYTHON, Wendy. / A validated enantioselective LC-MS/MS assay for quantification of a major chiral metabolite of an achiral 11-β-hydroxysteroid-dehydrogenase 1 inhibitor in human plasma : Application to a clinical pharmacokinetic study. Financial Crimes: Psychological, Technological, and Ethical Issues. editor / Michel Dion ; David Weisstub ; Jean-Loup Richet. Vol. 1022 Switzerland : Springer, 2016. pp. 167-172 (Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences).
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title = "A validated enantioselective LC-MS/MS assay for quantification of a major chiral metabolite of an achiral 11-β-hydroxysteroid-dehydrogenase 1 inhibitor in human plasma: Application to a clinical pharmacokinetic study",
abstract = "BMS-823778 is a potent 11-β-hydroxysteroid-dehydrogenase 1 (11βHSD-1) inhibitor and a potential therapeutic agent for type 2 diabetes mellitus (T2DM). A high performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay was developed and validated to enable reliable separation and quantification of both enantiomers of a chiral hydroxy metabolite (BMT-094817) in human plasma. Following liquid-liquid extraction in a 96-well plate format, chromatographic separation of the metabolite enantiomers was achieved by isocratic elution on a Chiralpak IA-3 column. Chromatographic conditions were optimized to ensure separation of both metabolite enantiomers. Metabolite enantiomers and stable isotope-labeled (SIL) internal standards were detected by positive ion electrospray tandem mass spectrometry. The LC-MS/MS assay was validated over a concentration range of 0.200-200 ng/mL. Intra- and inter-assay precision values for replicate quality control samples were less than 9.9{\%} for both enantiomers during the assay validation. Mean quality control accuracy values were within ±7.3{\%}. Assay recoveries were high (>75{\%}) and consistent across the assay range. The metabolite enantiomers were stable in human blood for 2 h on ice. The analytes were also stable in human plasma for 25 h at room temperature, 34 days at -20 °C and -70 °C, and following five freeze-thaw cycles. No interconversion of the metabolite enantiomers was detected under any bioanalytical stress conditions, from blood collection/processing through extracted sample storage. The validated assay was successfully applied to the quantification of both metabolite enantiomers in human plasma in support of a human pharmacokinetic study.",
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ARNOLD, B & BONYTHON, W 2016, A validated enantioselective LC-MS/MS assay for quantification of a major chiral metabolite of an achiral 11-β-hydroxysteroid-dehydrogenase 1 inhibitor in human plasma: Application to a clinical pharmacokinetic study. in M Dion, D Weisstub & J-L Richet (eds), Financial Crimes: Psychological, Technological, and Ethical Issues. vol. 1022, Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences, Springer, Switzerland, pp. 167-172. https://doi.org/10.1016/j.jchromb.2016.03.043

A validated enantioselective LC-MS/MS assay for quantification of a major chiral metabolite of an achiral 11-β-hydroxysteroid-dehydrogenase 1 inhibitor in human plasma : Application to a clinical pharmacokinetic study. / ARNOLD, Bruce; BONYTHON, Wendy.

Financial Crimes: Psychological, Technological, and Ethical Issues. ed. / Michel Dion; David Weisstub; Jean-Loup Richet. Vol. 1022 Switzerland : Springer, 2016. p. 167-172 (Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences).

Research output: A Conference proceeding or a Chapter in BookChapter

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T1 - A validated enantioselective LC-MS/MS assay for quantification of a major chiral metabolite of an achiral 11-β-hydroxysteroid-dehydrogenase 1 inhibitor in human plasma

T2 - Application to a clinical pharmacokinetic study

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AU - BONYTHON, Wendy

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N2 - BMS-823778 is a potent 11-β-hydroxysteroid-dehydrogenase 1 (11βHSD-1) inhibitor and a potential therapeutic agent for type 2 diabetes mellitus (T2DM). A high performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay was developed and validated to enable reliable separation and quantification of both enantiomers of a chiral hydroxy metabolite (BMT-094817) in human plasma. Following liquid-liquid extraction in a 96-well plate format, chromatographic separation of the metabolite enantiomers was achieved by isocratic elution on a Chiralpak IA-3 column. Chromatographic conditions were optimized to ensure separation of both metabolite enantiomers. Metabolite enantiomers and stable isotope-labeled (SIL) internal standards were detected by positive ion electrospray tandem mass spectrometry. The LC-MS/MS assay was validated over a concentration range of 0.200-200 ng/mL. Intra- and inter-assay precision values for replicate quality control samples were less than 9.9% for both enantiomers during the assay validation. Mean quality control accuracy values were within ±7.3%. Assay recoveries were high (>75%) and consistent across the assay range. The metabolite enantiomers were stable in human blood for 2 h on ice. The analytes were also stable in human plasma for 25 h at room temperature, 34 days at -20 °C and -70 °C, and following five freeze-thaw cycles. No interconversion of the metabolite enantiomers was detected under any bioanalytical stress conditions, from blood collection/processing through extracted sample storage. The validated assay was successfully applied to the quantification of both metabolite enantiomers in human plasma in support of a human pharmacokinetic study.

AB - BMS-823778 is a potent 11-β-hydroxysteroid-dehydrogenase 1 (11βHSD-1) inhibitor and a potential therapeutic agent for type 2 diabetes mellitus (T2DM). A high performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay was developed and validated to enable reliable separation and quantification of both enantiomers of a chiral hydroxy metabolite (BMT-094817) in human plasma. Following liquid-liquid extraction in a 96-well plate format, chromatographic separation of the metabolite enantiomers was achieved by isocratic elution on a Chiralpak IA-3 column. Chromatographic conditions were optimized to ensure separation of both metabolite enantiomers. Metabolite enantiomers and stable isotope-labeled (SIL) internal standards were detected by positive ion electrospray tandem mass spectrometry. The LC-MS/MS assay was validated over a concentration range of 0.200-200 ng/mL. Intra- and inter-assay precision values for replicate quality control samples were less than 9.9% for both enantiomers during the assay validation. Mean quality control accuracy values were within ±7.3%. Assay recoveries were high (>75%) and consistent across the assay range. The metabolite enantiomers were stable in human blood for 2 h on ice. The analytes were also stable in human plasma for 25 h at room temperature, 34 days at -20 °C and -70 °C, and following five freeze-thaw cycles. No interconversion of the metabolite enantiomers was detected under any bioanalytical stress conditions, from blood collection/processing through extracted sample storage. The validated assay was successfully applied to the quantification of both metabolite enantiomers in human plasma in support of a human pharmacokinetic study.

KW - 11-β-Hydroxysteroid-dehydrogenase 1 inhibitor

KW - Assay

KW - Chiral separation

KW - LC-MS/MS

KW - Metabolite

KW - Pharmacokinetics

KW - 11-beta-Hydroxysteroid-dehydrogenase 1 inhibitor

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U2 - 10.1016/j.jchromb.2016.03.043

DO - 10.1016/j.jchromb.2016.03.043

M3 - Chapter

SN - 9783319324180

VL - 1022

T3 - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences

SP - 167

EP - 172

BT - Financial Crimes: Psychological, Technological, and Ethical Issues

A2 - Dion, Michel

A2 - Weisstub, David

A2 - Richet, Jean-Loup

PB - Springer

CY - Switzerland

ER -

ARNOLD B, BONYTHON W. A validated enantioselective LC-MS/MS assay for quantification of a major chiral metabolite of an achiral 11-β-hydroxysteroid-dehydrogenase 1 inhibitor in human plasma: Application to a clinical pharmacokinetic study. In Dion M, Weisstub D, Richet J-L, editors, Financial Crimes: Psychological, Technological, and Ethical Issues. Vol. 1022. Switzerland: Springer. 2016. p. 167-172. (Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences). https://doi.org/10.1016/j.jchromb.2016.03.043