title: Development and validation of a high-throughput assay for quantification of the proliferation inhibitor ABT-578 using LC/LC-MS/MS in blood and tissue samples.
abstract: We report here a specific, automated LC/LC-MS/MS assay for the quantification of ABT-578 in human and rabbit blood and rabbit tissues for drug-eluting stent development. After protein precipitation, samples were injected into the HPLC system and extracted online using a high flow of 5 mL/min. The extracts were then backflushed onto the analytic column. The [M+Na] of ABT-578 (m/z 988.6-->369.4) and its internal standard sirolimus (m/z 936.5-->409.3) were monitored. Extraction and analysis took 4 minutes. The assay was validated following the US Food & Drug Administration guidelines. Linearity was 0.025-25 ng/mL for most matrices. In human blood, interday accuracies were 81.8% (at 0.025 ng/mL), 91.0% (1 ng/mL), and 99.5% (50 ng/mL), and interday precisions were 10.7% (0.025 ng/mL), 3.0% (1 ng/mL), and 1.8% (50 ng/mL).
authors.affiliation: Clinical Research and Development, Department of Anesthesiology, University of Colorado Health Sciences Center, Denver, Colorado 80262, USA.
authors.forename: Yan Ling
authors.initials: YL
authors.lastname: Zhang
authors.forename: Jamie
authors.initials: J
authors.lastname: Bendrick-Peart
authors.forename: Tobin
authors.initials: T
authors.lastname: Strom
authors.forename: Manuel
authors.initials: M
authors.lastname: Haschke
authors.forename: Uwe
authors.initials: U
authors.lastname: Christians
mesh_terms: D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006131:Growth Inhibitors / Q000097:blood / Q000737:chemistry / Q000493:pharmacokinetics; D006801:Humans; D015394:Molecular Structure; D011817:Rabbits; D015203:Reproducibility of Results; D020123:Sirolimus / Q000031:analogs & derivatives* / Q000097:blood / Q000493:pharmacokinetics; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D013678:Technology, Pharmaceutical / Q000379:methods; D014018:Tissue Distribution
chemical_list: D006131:Growth Inhibitors; C489443:zotarolimus; D020123:Sirolimus