Integrated Biomimetic 2D-LC and Permeapad® Assay for Profiling the Transdermal Diffusion of Pharmaceutical Compounds

A comprehensive two-dimensional liquid chromatography platform (LC × LC) was developed and validated for dermal permeability studies. In this implementation, the two separation dimensions were applied to mimic the layered structure of human skin: a ceramide-like stationary phase in the first dimension (¹D) to simulate the lipid-rich epidermis, and an immobilized artificial membrane (IAM) phase in the second (²D) to emulate the dermis. Experimental conditions were optimised to reflect the microenvironment of the in vivo skin. For validation purposes, 43 pharmaceutical and cosmetic compounds whose transdermal permeability coefficients (log K_p) were known from the scientific literature were selected as model solutes. A good degree of separation was achieved across the whole dataset, and affinity profiles correlated with transdermal passage properties, suggesting that retention within specific chromatographic ranges may be predictive of skin permeation. To complement this approach, mass diffusion measurements were also conducted using Permeapad^® 96-well plates and LC was performed on a narrow bore column in MS-friendly conditions. These log K_p values were compared against both in vivo and chromatographic retention data. The combined use of these techniques offers a strategic framework for profiling new chemical entities for their dermal absorption in a manner that is both ethically compliant and eco-sustainable.