Application of separation methods for in vitro prediction of blood–brain barrier permeability—The state of the art

•The in vitro methods for the estimation of drugs penetration through BBB were discussed.•The application of separation methods for log BB prediction was reviewed.•Both chromatographic and electromigration techniques were found to feature high predictive potential. Despite many efforts, drug discovery pipeline is still a highly inefficient process. Nowadays, when combinatorial chemistry enables to synthesize hundreds of new drugs candidates, methods for rapid assessment of biopharmaceutical parameters of new compounds are highly desired. Over one-third of drugs candidates is rejected because of unsatisfactory pharmacokinetic properties. In the drug discovery process, the blood-brain barrier (BBB) permeability plays a critical role for central nervous system active drugs candidates as well as non-central nervous system active drugs. For this reason, knowledge on the BBB permeability of compounds is essential in the development of new medicines. The review was focused on the application of different separation methods for BBB permeability assessment. Both chromatographic and electrophoretic methods were described. In the article, the advantages and limitations of well-established chromatographic methods like immobilized artificial membrane chromatography or micellar liquid chromatography, and less common techniques were discussed. Special attention was devoted to methods were microemulsion is used as mobile or pseudostationary phases.