Jump to main content
Jump to site search
Access to RSC content Close the message box

Continue to access RSC content when you are not at your institution. Follow our step-by-step guide.


Issue 20, 2013
Previous Article Next Article

Correlation of in vitro and in vivo plasma protein binding using ultracentrifugation and UPLC-tandem mass spectrometry

Author affiliations

Abstract

The aim of the present study is to develop and demonstrate the correlation between in vitro and in vivo Plasma Protein Binding (PPB) using the ultracentrifugation method for its validation by using marketed compounds like atenolol, theophylline and phenytoin. In this study, in vitro PPB is carried out using ultracentrifugation, by spiking the selected marketed compounds at concentrations of 5 and 15 μM in plasma. In an in vivo study, rats (n = 3) were given a single oral dose (10 mg kg−1) and post-dose samples were subjected to ultracentrifugation to obtain the protein-free fraction. A rapid and highly sensitive method was developed and validated for determining the free fraction of marketed compounds in rat plasma using protein precipitation and analysis using an ultra performance liquid chromatography electrospray ionization (ESI) tandem mass spectrometer system (UPLC-MS/MS). The in vitro free fraction (fup) values were 0.93 ± 0.07 for atenolol, 0.31 ± 0.03 for theophylline and 0.09 ± 0.02 for phenytoin which correlated well with the corresponding in vivo values of 0.91 ± 0.03 for atenolol, 0.25 ± 0.02 for theophylline and 0.09 ± 0.01 for phenytoin with a coefficient of variation less than 11.06%, 11.45% and 13.67%, respectively. Therefore the validated high-throughput in vitro PPB study is expected to have a powerful impact on reducing the cost as well as time in the drug discovery process.

Graphical abstract: Correlation of in vitro and in vivo plasma protein binding using ultracentrifugation and UPLC-tandem mass spectrometry

Back to tab navigation

Supplementary files

Article information


Submitted
04 Jun 2013
Accepted
29 Jul 2013
First published
31 Jul 2013

Analyst, 2013,138, 6106-6116
Article type
Paper

Correlation of in vitro and in vivo plasma protein binding using ultracentrifugation and UPLC-tandem mass spectrometry

C. H. Srikanth, T. Chaira, S. Sampathi, S. V. B. and R. B. Bambal, Analyst, 2013, 138, 6106
DOI: 10.1039/C3AN01116J

Social activity

Search articles by author

Spotlight

Advertisements