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Updated: Aug 30, 2026
A Plasma Sample Preparation for Mass Spectrometry using an Automated Workstation
Published on: April 24, 2020
Semi-automatic high-throughput determination of plasma protein binding using a 96-well plate filtrate assembly and
Eliza N Fung1, Yung-Hsiang Chen, Yau Yi Lau
1Schering-Plough Research Institute, Drug Metabolism and Pharmacokinetics, 2015 Galloping Hill Road, K-15-2-2700 Kenilworth, NJ 07033, USA.
Abstract:
A semi-automatic, high-throughput method has been developed to rapidly assess plasma protein binding of new chemical entities in drug discovery phase. New chemical entities are mixed with plasma and the unbound fractions are separated from the bound fraction by ultrafiltration in a 96-well filtrate assembly. The unbound fractions are then analyzed by fast liquid chromatography-tandem mass spectrometry (LC-MS/MS). Sample handling is automated by a robotic system. Employing a cocktail approach where multiple new chemical entities are allowed to bind to plasma proteins in the same well has further increased the throughput. We have validated the method with 12 commercially available compounds. The plasma protein binding data obtained by this method are comparable with the literature values. This method enables the determination of protein binding for 32 compounds in one single experiment instead of 1-2 compounds using the conventional methods.
