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Development and validation of an LC-MS/MS generic assay platform for small molecule drug bioanalysis

  1. Author:
    Parise, Robert A
    Covey, Joseph M
    Hollingshead,Melinda
    Srivastava,Apurva
    Synold, Timothy W
    Beumer, Jan H
  2. Author Address

    Cancer Therapeutics Program, UPMC Hillman Cancer Center, Pittsburgh, PA, United States. Electronic address: parisera@upmc.edu., Developmental Therapeutics Program, Division of Cancer Treatment and Diagnosis, National Cancer Institute, Bethesda, MD, United States. Electronic address: coveyj@dtpepn.nci.nih.gov., Biological Testing Branch, Developmental Therapeutics Program, Frederick National Laboratory for Cancer Research, Frederick, MD, United States. Electronic address: hollingm@mail.nih.gov., Clinical Pharmacodynamics Biomarker Program, Applied/Developmental Research Directorate, Leidos Biomedical Research, Inc., Frederick National Laboratory for Cancer Research, Frederick, MD, United States. Electronic address: srivastavaa4@mail.nih.gov., Department of Medical Oncology and Therapeutics Research, City of Hope Comprehensive Cancer Center, Duarte, CA, United States. Electronic address: TSynold@coh.org., Cancer Therapeutics Program, UPMC Hillman Cancer Center, Pittsburgh, PA, United States; Department of Pharmaceutical Sciences, School of Pharmacy, University of Pittsburgh, Pittsburgh, PA, United States; Division of Hematology-Oncology, Department of Medicine, University of Pittsburgh School of Medicine, Pittsburgh, PA, United States. Electronic address: beumerj@gmail.com.,
    1. Year: 2021
    2. Date: Sep 5
    3. Epub Date: 2021 06 02
  1. Journal: Journal of pharmaceutical and biomedical analysis
    1. 203
    2. Pages: 114185
  2. Type of Article: Article
  3. Article Number: 114185
  4. ISSN: 0731-7085
  1. Abstract:

    We developed a generic high-performance liquid chromatography mass spectrometry approach for quantitation of small molecule compounds without availability of isotopically labelled standard. The assay utilized 50 µL of plasma and offers 8 potential internal standards (IS): acetaminophen, veliparib, busulfan, neratinib, erlotinib, abiraterone, bicalutamide, and paclitaxel. Preparation consisted of acetonitrile protein precipitation and aqueous dilution in a 96 well-plate format. Chromatographic separation was achieved with a Kinetex C18 reverse phase (2.6 µm, 2 mm x 50 mm) column and a gradient of 0.1 % formic acid in acetonitrile and water over an 8 min run time. Mass spectrometric detection was performed on an AB SCIEX4000QTRAP with electrospray, positive-mode ionization. Performance of the generic approach was evaluated with seven drugs (LMP744, olaparib, cabozantinib, triapine, ixabepilone, berzosertib, eribulin) for which validated assays were available. The 8 IS covered a range of polarity, size, and ionization; eluted over the range of chromatographic retention times; were quantitatively extracted; and suffered limited matrix effects. The generic approach proved to be linear for test drugs evaluated over at least 3 orders of magnitude starting at 1-10 ng/mL, with extension of assay ranges with analyte isotopologue MRM channels. At a bias of less than 16 % and precision within 15 %, the assay performance was acceptable. The generic approach has become a useful tool to further define the pharmacology of drugs studied in our laboratory and may be utilized as described, or as starting point to develop drug-specific assays with more extensive performance characterization. Copyright © 2021 Elsevier B.V. All rights reserved.

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External Sources

  1. DOI: 10.1016/j.jpba.2021.114185
  2. PMID: 34111734
  3. WOS: 000679331300011
  4. PII : S0731-7085(21)00296-X

Library Notes

  1. Fiscal Year: FY2020-2021
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