How to define the experimental conditions of skin permeation assays for drugs presenting biopharmaceutical limitations? The experience with testosterone.

Cutaneous permeation assays are crucial to attest the performance or bioequivalence of topical or transdermal products. Although the official guidelines (e.g., FDA/EMA) play a key role in harmonizing the experimental design, alternative methods are often proposed by the scientific community, which makes it difficult to compare results from different studies. In this review, permeation assays with testosterone (TST) were selected to show this high variability in drug transport rate. The main sources of variation discussed were tissue thickness, animal model, donor and receptor fluid constitution, type of solubilizing agent used in aqueous fluids, drug concentration, degree of supersaturation, skin lipid content, number of experimental times and the physical-chemical stability of the molecule in test fluids. This variation becomes even more critical for molecules that present biopharmaceutical limitations such as TST. In addition, the skin presents specific receptors for this hormone due to its physiological action in this region of the body, which makes the evaluation of the TST transport rate in this tissue even more challenging. The impact of each experimental parameter mentioned above on the flux or permeation coefficient of TST is discussed in detail in the review. Assays used to evaluate tissue integrity are also presented.