Screening application of the miniaturized and automated human BBB in vitro model. (A) Values (mean ± SD) of the unbound fraction ratio of the CSF/plasma in humans (in vivo Kp,uu,CSF) data obtained from Friden M et al. 2009. Unbound fraction ratios of the brain/plasma predicted in vitro (in vitro Kp,uu,brain) through the 96 TW miniaturized and automated human models. The concentrations were detected by mass spectrometry (LC–MS/MS system) with the calculation of the total amount of unbound molecules recovered (recovery, expressed in percentage). (B) Correlation between the Kp,uu,CSF and Kp,uu,brain nonlinear equations; R2 = 0.8808; n = 3; N = 1.