Hydrophilic interaction chromatography/tandem mass spectrometry for the simultaneous determination of three polar non-structurally related compounds, imipenem, cilastatin and an investigational beta-lactamase inhibitor, MK-4698, in biological matrices.
A method coupling hydrophilic interaction chromatography (HILIC) with tandem mass spectrometry (MS/MS) has been developed for the simultaneous determination of three polar non-structurally related compounds--a carbapenem antibiotic, imipenem (IMP), a renal dehydropeptidase inhibitor, cilastatin (CIL), and an investigational beta-lactamase inhibitor, MK-4698 (BLI), in rat plasma, monkey plasma and mouse blood. The analytes were extracted through protein precipitation, chromatographed on a Waters Atlantis HILIC column, and detected on a Sciex API4000 mass spectrometer using a Turbo-Ion Spray ion source in positive ionization mode following multiple-reaction monitoring (MRM). The assay dynamic range was 0.1-100 microg/mL for IMP, CIL and BLI, respectively, using a total of 20-25 microL biologic samples, and the total HPLC/MS/MS run time was 4 min/injection. The assay was found to be sensitive, selective and reproducible. The challenges, namely, sample stability, blood sample processing, matrix effect in monkey study samples, and dilution re-assays for the limited mouse blood samples, are resolved and discussed. This technique allowed rapid analysis of polar compounds in biologic matrixes with satisfactory chromatographic retention and increased throughput.
Xu
Yang
Y
Merck Research Laboratories, Department of Drug Metabolism and Pharmacokinetics, 770 Sumneytown Pike, WP75B-300, West Point, PA 19486, USA. yang_xu@merck.com
Xie
Wei
W
Miller-Stein
Cynthia M
CM
Woolf
Eric J
EJ
D000818:Animals; D000900:Anti-Bacterial Agents / Q000097:blood / Q000737:chemistry; D002845:Chromatography / Q000379:methods*; D015377:Cilastatin / Q000097:blood / Q000737:chemistry*; D004791:Enzyme Inhibitors / Q000097:blood / Q000737:chemistry*; D000882:Haplorhini; D015378:Imipenem / Q000097:blood / Q000737:chemistry*; D051379:Mice; D051381:Rats; D053719:Tandem Mass Spectrometry / Q000379:methods*; D065093:beta-Lactamase Inhibitors
D000900:Anti-Bacterial Agents; D004791:Enzyme Inhibitors; D065093:beta-Lactamase Inhibitors; D015377:Cilastatin; D015378:Imipenem