Robert A Parise1, Joseph M Covey2, Melinda G Hollingshead3, Apurva K Srivastava4, Timothy W Synold5, Jan H Beumer6. 1. Cancer Therapeutics Program, UPMC Hillman Cancer Center, Pittsburgh, PA, United States. Electronic address: parisera@upmc.edu. 2. Developmental Therapeutics Program, Division of Cancer Treatment and Diagnosis, National Cancer Institute, Bethesda, MD, United States. Electronic address: coveyj@dtpepn.nci.nih.gov. 3. Biological Testing Branch, Developmental Therapeutics Program, Frederick National Laboratory for Cancer Research, Frederick, MD, United States. Electronic address: hollingm@mail.nih.gov. 4. 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. 5. Department of Medical Oncology and Therapeutics Research, City of Hope Comprehensive Cancer Center, Duarte, CA, United States. Electronic address: TSynold@coh.org. 6. 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.
Abstract
AIM: We developed a generic high-performance liquid chromatography mass spectrometry approach for quantitation of small molecule compounds without availability of isotopically labelled standard. METHODS: 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. RESULTS: 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. CONCLUSION: 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.
AIM: We developed a generic high-performance liquid chromatography mass spectrometry approach for quantitation of small molecule compounds without availability of isotopically labelled standard. METHODS: 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. RESULTS: 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. CONCLUSION: 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.
Authors: Robert A Parise; Julianne L Holleran; Jan H Beumer; Suresh Ramalingam; Merrill J Egorin Journal: J Chromatogr B Analyt Technol Biomed Life Sci Date: 2006-05-24 Impact factor: 3.205
Authors: Kyu-Pyo Kim; Robert A Parise; Julianne L Holleran; Lionel D Lewis; Leonard Appleman; Nielka van Erp; Michael J Morris; Jan H Beumer Journal: J Pharm Biomed Anal Date: 2017-02-13 Impact factor: 3.935
Authors: Brian F Kiesel; Robert A Parise; Alvin Wong; Kiana Keyvanjah; Samuel Jacobs; Jan H Beumer Journal: J Pharm Biomed Anal Date: 2016-11-22 Impact factor: 3.935
Authors: Robert A Parise; Mohammad Shawaqfeh; Merrill J Egorin; Jan H Beumer Journal: J Chromatogr B Analyt Technol Biomed Life Sci Date: 2008-07-31 Impact factor: 3.205
Authors: Brian F Kiesel; Jonas Scemama; Robert A Parise; Liza Villaruz; Andre Iffland; Austin Doyle; Percy Ivy; Edward Chu; Christopher J Bakkenist; Jan H Beumer Journal: J Pharm Biomed Anal Date: 2017-08-31 Impact factor: 3.935
Authors: C M Nijenhuis; L Lucas; H Rosing; J H M Schellens; J H Beijnen Journal: J Chromatogr B Analyt Technol Biomed Life Sci Date: 2013-09-23 Impact factor: 3.205
Authors: Robert A Parise; Merrill J Egorin; Julie L Eiseman; Erin Joseph; Joseph M Covey; Jan H Beumer Journal: Rapid Commun Mass Spectrom Date: 2007 Impact factor: 2.419
Authors: Brian F Kiesel; Robert A Parise; Jette Tjørnelund; Mette K Christensen; Einars Loza; Hussein Tawbi; Edward Chu; Shivaani Kummar; Jan H Beumer Journal: J Pharm Biomed Anal Date: 2013-04-09 Impact factor: 3.935