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Single run measurements of drug‐protein binding by high‐performance frontal analysis capillary electrophoresis and mass spectrometry
Author(s) -
Wan Hong,
Östlund Åsa,
Jönsson Stefan,
Lindberg Walter
Publication year - 2005
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.1958
Subject(s) - chemistry , chromatography , free fraction , mass spectrometry , capillary electrophoresis , analytical chemistry (journal) , calibration curve , plasma protein binding , detection limit , biochemistry
A novel drug‐protein binding measurement method based on high‐performance frontal analysis and capillary electrophoresis (HPFA/CE) is presented. A single run measurement approach is proposed to circumvent utilization of a calibration curve that is often performed with HPFA. A sensitive mass spectrometer is applied as a detector enabling the measurement of in vitro protein binding at lower drug concentrations. Unbound free fraction and binding constants can be determined by a single run measurement by consecutive injections of an internal drug standard, a buffer plug and a drug‐protein mixture. Effects of injection volumes on peak height and plateau profile were investigated in two different separation systems, non‐volatile buffer and volatile buffer, with UV and mass spectrometry detection, respectively. A simplified one‐to‐one binding model is employed to evaluate the proposed method by using both single and multiple drug concentrations to measure the unbound free fraction and calculate the binding constants of some selected compounds. The method is suitable for rapid and direct screening of the binding of a drug to a specific protein or drug‐plasma protein binding. Copyright © 2005 John Wiley & Sons, Ltd.