| High-resolution isotope-dilution mass spectrometry using metabolism of isotope-labeled compounds: Application to drug metabolites. | |
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MedLine Citation:
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PMID: 23059872 Owner: NLM Status: In-Data-Review |
Abstract/OtherAbstract:
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RATIONALE: Herein we describe a generic quantitative method using high-resolution, isotope-dilution (HRID) metabolism of isotope-labeled compounds and apply it to the analysis of drug metabolites (DMs) in human plasma. Metabolites (drug) in Safety Testing (MIST) application was one goal. METHODS: Testosterone (T) and diclofenac (D) were chosen for mass defect characteristics. T, [(14) C]T, [(13) C(3) ]T, D, [(14) C]D, and [(13) C(6) ]D were metabolized separately in vitro to produce test metabolites. Liquid chromatography/radioactivity monitoring (LC/RAM) analysis was used to determine the concentration of the test metabolites in the incubates. The incubates containing 6β-hydroxy-T (6βHT), [(13) C(3) ]6βHT, 4'-hydroxy-D (4'HD) and [(13) C(6) ]4'HD were used to make standard curves. Plasma samples were prepared by 'dilute-and-shoot' and analyzed by LC/MS using SCIEX 5000 and Thermo Orbitrap instrumentation. RESULTS: Human hepatic microsomes and the S9 fraction produced between 2-6 μM β-hydroxy-T and 4'-hydroxy-D at 60 min starting with 10 μM parent drug as determined by LC/RAM. It was assumed that the amounts of [(13) C(3) ]6βHT and [(13) C(6) ]4'HD produced were similar. Dilutions and standard curves were prepared in human plasma. Analysis of the DMs by LC/MS/MS and LC/HRMS exhibited linear responses over a useable range. CONCLUSIONS: HRID with metabolism of an isotope-labeled compound reduces the number of analytical variables considerably. Metabolism of the parent drug to DMs represents a simpler alternative quantitative method compared with traditional approaches. The method will have useful applications for evaluating MIST situations. Copyright © 2012 John Wiley & Sons, Ltd. |
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Authors:
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J James Vrbanac; Allen Hilgers; Tyson Dubnicka; F Barclay Shilliday; David Humphries; Roger N Hayes |
Publication Detail:
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Type: Journal Article |
Journal Detail:
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Title: Rapid communications in mass spectrometry : RCM Volume: 26 ISSN: 1097-0231 ISO Abbreviation: Rapid Commun. Mass Spectrom. Publication Date: 2012 Nov |
Date Detail:
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Created Date: 2012-10-12 Completed Date: - Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 8802365 Medline TA: Rapid Commun Mass Spectrom Country: England |
Other Details:
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Languages: eng Pagination: 2569-76 Citation Subset: IM |
Copyright Information:
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Copyright © 2012 John Wiley & Sons, Ltd. |
Affiliation:
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Discovery Sciences, MPI Research, Mattawan, MI, 49071, USA. jim.vrbanac@mpiresearch.com. |
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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