title: A simplified and completely automated workflow for regulated LC-MS/MS bioanalysis using cap-piercing direct sampling and evaporation-free solid phase extraction.
abstract: Automated sample extraction for regulated bioanalysis by liquid chromatography/tandem mass spectrometry (LC-MS/MS) still presents significant challenges. A new sample preparation methodology with a simplified and completely automated workflow was developed to overcome these challenges using cap piercing for direct biofluid transfer and evaporation-free solid phase extraction (SPE). Using pierceable cap sample tubes, a robotic liquid handler was able to sample without uncapping or recapping during sample preparation. Evaporation for SPE was eliminated by using a mobile phase-compatible elution solvent followed by sample dilution prior to LC-MS/MS analysis. Presented here are three LC-MS/MS assays validated using this methodology to support three CNS drug development programs: (1) BMS-763534 and its metabolite, BMS-790318, in dog plasma; (2) BMS-694153 in monkey plasma; and (3) Pexacerfont (BMS-562086) and two metabolites, BMS-749241 and DPH-123554, in human plasma. These assays were linear from 1.00 to 1000 or 2.00 to 2000ng/mL for each analyte with excellent assay accuracy, precision and reproducibility. These assays met acceptance criteria for regulated bioanalysis and have been successfully applied to drug development study samples. The methodology described here successfully eliminated all manual intervention steps achieving fully automated sample preparation without compromising assay performance. Importantly, this methodology eliminates the potential exposure of the bioanalyst to any infectious biofluids during sample preparation.
authors: Analytical & Bioanalytical Development, Bristol-Myers Squibb Company, Princeton, NJ 08540, United States.
authors: Naiyu
authors: N
authors: Zheng
authors: Adela
authors: A
authors: Buzescu
authors: Stephanie
authors: S
authors: Pasas-Farmer
authors: Mark E
authors: ME
authors: Arnold
authors: Zheng
authors: Z
authors: Ouyang
authors: Mohammed
authors: M
authors: Jemal
authors: Qianping
authors: Q
authors: Peng
authors: Terry
authors: T
authors: Van Vleet
authors: Jianing
authors: J
authors: Zeng
mesh_terms: D000818:Animals; D000077221:Calcitonin Gene-Related Peptide Receptor Antagonists; D002853:Chromatography, Liquid / Q000191:economics / Q000295:instrumentation / Q000379:methods; D004285:Dogs; D004867:Equipment Design; D000882:Haplorhini; D057166:High-Throughput Screening Assays / Q000191:economics / Q000295:instrumentation / Q000379:methods; D006801:Humans; D007191:Indazoles / Q000097:blood* / Q000378:metabolism; D011720:Pyrazoles / Q000097:blood* / Q000378:metabolism; D052999:Quinazolinones / Q000097:blood* / Q000378:metabolism; D018019:Receptors, Corticotropin-Releasing Hormone / Q000037:antagonists & inhibitors*; D012680:Sensitivity and Specificity; D052616:Solid Phase Extraction / Q000191:economics / Q000295:instrumentation / Q000379:methods*; D013048:Specimen Handling / Q000191:economics / Q000295:instrumentation / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000191:economics / Q000295:instrumentation / Q000379:methods*; D014227:Triazines / Q000097:blood* / Q000378:metabolism; D000099052:CRF Receptor, Type 1
chemical_list: C531228:4-(8-fluoro-2-oxo-1,2-dihydroquinazolin-3(4H)-yl)-N-(3-(7-methyl-1H-indazol-5-yl)-1-oxo-1-(4-(piperidin-1-yl)piperidin-1-yl)propan-2-yl)piperidine-1-carboxamide; D000077221:Calcitonin Gene-Related Peptide Receptor Antagonists; D007191:Indazoles; D011720:Pyrazoles; D052999:Quinazolinones; D018019:Receptors, Corticotropin-Releasing Hormone; D014227:Triazines; D000099052:CRF Receptor, Type 1; C542342:pexacerfont