Development and bioanalytical validation of a liquid chromatographic-tandem mass spectrometric (LC-MS/MS) method for the quantification of the CCR5 antagonist maraviroc in human plasma

Joshua F. Emory, Lauren A. Seserko, Mark A Marzinke

Research output: Contribution to journalArticle

Abstract

Background: Maraviroc is a CCR5 antagonist that has been utilized as a viral entry inhibitor in the management of HIV-1. Current clinical trials are pursuing maraviroc drug efficacy in both oral and topical formulations. Therefore, in order to fully understand drug pharmacokinetics, a sensitive method is required to quantify plasma drug concentrations. Methods: Maraviroc-spiked plasma was combined with acetonitrile containing an isotopically-labeled internal standard, and following protein precipitation, samples were evaporated to dryness and reconstituted for liquid chromatographic-tandem mass spectrometric (LC-MS/MS) analysis. Chromatographic separation was achieved on a Waters BEH C8, 50. ×. 2.1. mm UPLC column, with a 1.7. μm particle size and the eluent was analyzed using an API 4000 mass analyzer in selected reaction monitoring mode. The method was validated as per FDA Bioanalytical Method Validation guidelines. Results: The analytical measuring range of the LC-MS/MS method is 0.5-1000ng/ml. Calibration curves were generated using weighted 1/x2 quadratic regression. Inter-and intra-assay precision was ≤5.38% and ≤5.98%, respectively; inter-and intra-assay accuracy (%DEV) was ≤10.2% and ≤8.44%, respectively. Additional studies illustrated similar matrix effects between maraviroc and its internal standard, and that maraviroc is stable under a variety of conditions. Method comparison studies with a reference LC-MS/MS method show a slope of 0.948 with a Spearman coefficient of 0.98. Conclusions: Based on the validation metrics, we have generated a sensitive and automated LC-MS/MS method for maraviroc quantification in human plasma.

Original languageEnglish (US)
Pages (from-to)198-205
Number of pages8
JournalClinica Chimica Acta
Volume431
DOIs
StatePublished - Apr 20 2014

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Plasma (human)
Liquids
Assays
Pharmaceutical Preparations
Plasmas
Pharmacokinetics
Application programming interfaces (API)
maraviroc
Particle size
Particle Size
Calibration
HIV-1
Water
Monitoring
Clinical Trials
Guidelines
Proteins

Keywords

  • Assay validation
  • CCR5 antagonist
  • HIV
  • LC-MS/MS
  • Maraviroc

ASJC Scopus subject areas

  • Biochemistry
  • Clinical Biochemistry
  • Biochemistry, medical
  • Medicine(all)

Cite this

@article{9e0ae4f803aa4a71b6b7b05aed1f7991,
title = "Development and bioanalytical validation of a liquid chromatographic-tandem mass spectrometric (LC-MS/MS) method for the quantification of the CCR5 antagonist maraviroc in human plasma",
abstract = "Background: Maraviroc is a CCR5 antagonist that has been utilized as a viral entry inhibitor in the management of HIV-1. Current clinical trials are pursuing maraviroc drug efficacy in both oral and topical formulations. Therefore, in order to fully understand drug pharmacokinetics, a sensitive method is required to quantify plasma drug concentrations. Methods: Maraviroc-spiked plasma was combined with acetonitrile containing an isotopically-labeled internal standard, and following protein precipitation, samples were evaporated to dryness and reconstituted for liquid chromatographic-tandem mass spectrometric (LC-MS/MS) analysis. Chromatographic separation was achieved on a Waters BEH C8, 50. ×. 2.1. mm UPLC column, with a 1.7. μm particle size and the eluent was analyzed using an API 4000 mass analyzer in selected reaction monitoring mode. The method was validated as per FDA Bioanalytical Method Validation guidelines. Results: The analytical measuring range of the LC-MS/MS method is 0.5-1000ng/ml. Calibration curves were generated using weighted 1/x2 quadratic regression. Inter-and intra-assay precision was ≤5.38{\%} and ≤5.98{\%}, respectively; inter-and intra-assay accuracy ({\%}DEV) was ≤10.2{\%} and ≤8.44{\%}, respectively. Additional studies illustrated similar matrix effects between maraviroc and its internal standard, and that maraviroc is stable under a variety of conditions. Method comparison studies with a reference LC-MS/MS method show a slope of 0.948 with a Spearman coefficient of 0.98. Conclusions: Based on the validation metrics, we have generated a sensitive and automated LC-MS/MS method for maraviroc quantification in human plasma.",
keywords = "Assay validation, CCR5 antagonist, HIV, LC-MS/MS, Maraviroc",
author = "Emory, {Joshua F.} and Seserko, {Lauren A.} and Marzinke, {Mark A}",
year = "2014",
month = "4",
day = "20",
doi = "10.1016/j.cca.2014.02.008",
language = "English (US)",
volume = "431",
pages = "198--205",
journal = "Clinica Chimica Acta",
issn = "0009-8981",
publisher = "Elsevier",

}

TY - JOUR

T1 - Development and bioanalytical validation of a liquid chromatographic-tandem mass spectrometric (LC-MS/MS) method for the quantification of the CCR5 antagonist maraviroc in human plasma

AU - Emory, Joshua F.

AU - Seserko, Lauren A.

AU - Marzinke, Mark A

PY - 2014/4/20

Y1 - 2014/4/20

N2 - Background: Maraviroc is a CCR5 antagonist that has been utilized as a viral entry inhibitor in the management of HIV-1. Current clinical trials are pursuing maraviroc drug efficacy in both oral and topical formulations. Therefore, in order to fully understand drug pharmacokinetics, a sensitive method is required to quantify plasma drug concentrations. Methods: Maraviroc-spiked plasma was combined with acetonitrile containing an isotopically-labeled internal standard, and following protein precipitation, samples were evaporated to dryness and reconstituted for liquid chromatographic-tandem mass spectrometric (LC-MS/MS) analysis. Chromatographic separation was achieved on a Waters BEH C8, 50. ×. 2.1. mm UPLC column, with a 1.7. μm particle size and the eluent was analyzed using an API 4000 mass analyzer in selected reaction monitoring mode. The method was validated as per FDA Bioanalytical Method Validation guidelines. Results: The analytical measuring range of the LC-MS/MS method is 0.5-1000ng/ml. Calibration curves were generated using weighted 1/x2 quadratic regression. Inter-and intra-assay precision was ≤5.38% and ≤5.98%, respectively; inter-and intra-assay accuracy (%DEV) was ≤10.2% and ≤8.44%, respectively. Additional studies illustrated similar matrix effects between maraviroc and its internal standard, and that maraviroc is stable under a variety of conditions. Method comparison studies with a reference LC-MS/MS method show a slope of 0.948 with a Spearman coefficient of 0.98. Conclusions: Based on the validation metrics, we have generated a sensitive and automated LC-MS/MS method for maraviroc quantification in human plasma.

AB - Background: Maraviroc is a CCR5 antagonist that has been utilized as a viral entry inhibitor in the management of HIV-1. Current clinical trials are pursuing maraviroc drug efficacy in both oral and topical formulations. Therefore, in order to fully understand drug pharmacokinetics, a sensitive method is required to quantify plasma drug concentrations. Methods: Maraviroc-spiked plasma was combined with acetonitrile containing an isotopically-labeled internal standard, and following protein precipitation, samples were evaporated to dryness and reconstituted for liquid chromatographic-tandem mass spectrometric (LC-MS/MS) analysis. Chromatographic separation was achieved on a Waters BEH C8, 50. ×. 2.1. mm UPLC column, with a 1.7. μm particle size and the eluent was analyzed using an API 4000 mass analyzer in selected reaction monitoring mode. The method was validated as per FDA Bioanalytical Method Validation guidelines. Results: The analytical measuring range of the LC-MS/MS method is 0.5-1000ng/ml. Calibration curves were generated using weighted 1/x2 quadratic regression. Inter-and intra-assay precision was ≤5.38% and ≤5.98%, respectively; inter-and intra-assay accuracy (%DEV) was ≤10.2% and ≤8.44%, respectively. Additional studies illustrated similar matrix effects between maraviroc and its internal standard, and that maraviroc is stable under a variety of conditions. Method comparison studies with a reference LC-MS/MS method show a slope of 0.948 with a Spearman coefficient of 0.98. Conclusions: Based on the validation metrics, we have generated a sensitive and automated LC-MS/MS method for maraviroc quantification in human plasma.

KW - Assay validation

KW - CCR5 antagonist

KW - HIV

KW - LC-MS/MS

KW - Maraviroc

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U2 - 10.1016/j.cca.2014.02.008

DO - 10.1016/j.cca.2014.02.008

M3 - Article

C2 - 24561264

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VL - 431

SP - 198

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JO - Clinica Chimica Acta

JF - Clinica Chimica Acta

SN - 0009-8981

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