title: A Comparison of LC-MS/MS and a Fully Integrated Autosampler/Solid-Phase Extraction System for the Analysis of Protein Binding Samples.
abstract: A new analysis approach was evaluated for measuring plasma protein binding (PPB) of small molecules using the Agilent RapidFire high-throughput system coupled with a Sciex API 4000 mass spectrometer (RF-MS/MS). Thirty-three proprietary and 12 literature compounds were subjected to rapid equilibrium dialysis (RED) and evaluated in parallel using RF-MS/MS at 16.4 s/sample and traditional liquid chromatography-tandem mass spectrometry (LC-MS/MS) at 3.5 min/sample, thus making the RF-MS/MS analysis over 12 times faster than LC-MS/MS. The high-throughput analysis method that was developed demonstrated excellent correlation with the traditional LC-MS/MS analysis method with an r(2) value of 0.96. The RF-MS/MS analysis method was implemented to increase sample throughput, decrease turnaround time for PPB data, and decrease time burden on existing LC-MS/MS instruments.
authors.affiliation: Drug Metabolism & Pharmacokinetics, Biogen, Inc., Cambridge, MA, USA.
authors.forename: Adam
authors.initials: A
authors.lastname: Amaral
authors.affiliation: Analytical Sciences & Imaging, Novartis Institutes for Biomedical Research, Inc., Cambridge, MA, USA saran.chitra@novartis.com.
authors.forename: Chitra
authors.initials: C
authors.lastname: Saran
authors.affiliation: Drug Metabolism & Pharmacokinetics, Agios Pharmaceuticals, Cambridge, MA 02139 USA.
authors.forename: Jakal
authors.initials: J
authors.lastname: Amin
authors.affiliation: Drug Metabolism & Pharmacokinetics, Novartis Institutes for Biomedical Research, Inc., East Hanover, NJ, USA.
authors.forename: Panos
authors.initials: P
authors.lastname: Hatsis
mesh_terms: D002853:Chromatography, Liquid / Q000379:methods*; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D011485:Protein Binding; D011506:Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry*; D052616:Solid Phase Extraction; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D011506:Proteins
keywords: RapidFire; high throughput; mass spectrometry; protein binding