Increasing the Robustness of Biopharmaceutical Precipitation Assays - Part II: Recommendations on the use of FaSSIF.

Biopharmaceutical precipitation assays are an important in vitro tool to characterize the precipitation behavior of weakly basic drugs during their transit from the stomach into the small intestine. To mimic the intestinal fluids more closely, biorelevant media like Fasted State Simulated Intestinal Fluid (FaSSIF) and versions thereof are often applied. When applying UV analytics to measure the drug concentration during the transfer experiments, changes in the UV spectrum of the medium have been observed when FaSSIF was stored over a longer period of time or under accelerated conditions. Therefore, this study aimed at evaluating the stability of FaSSIF under various storage conditions. Furthermore, the impact of stressed FaSSIF on the supersaturation and precipitation behavior of ketoconazole was investigated. As a result of this study, it was demonstrated that the FaSSIF powder composition changes during storage, which, in turn, impacts the supersaturation and precipitation behavior of ketoconazole in in vitro transfer studies. Based on the results of this study, we provide recommendations on the application of FaSSIF in biopharmaceutical precipitation assays with the aim to increase reproducibility and enhance data reliability for those compounds where changing FaSSIF composition may impact the supersaturation and precipitation behavior.

The effect of buffer species on biorelevant dissolution and precipitation assays - Comparison of phosphate and bicarbonate buffer.

Biorelevant solubility and dissolution testing is an important tool during pharmaceutical development, however, solubility experiments performed using biorelevant media often do not properly match the solubility data observed in human intestinal fluids. Even though the bicarbonate buffer is the predominant buffer system in the small intestine, in vitro assays are commonly performed using non-volatile buffer systems like phosphate and maleate. In the current study, bicarbonate- and phosphate-buffered biorelevant media were applied to solubility, dissolution, and precipitation testing for a broad range of model compounds. It was found that the medium affects primarily the dissolution kinetics. However, with the knowledge of the unique buffering properties of bicarbonate buffer in the diffusion layer, it was not always possible to predict the effect of buffer species on solubility and dissolution when changing from phosphate to bicarbonate buffer. This once again highlights the special role of bicarbonate buffer for simulating the conditions in the human intestinal fluids. Moreover, it is necessary to further investigate the factors which may cause the differences in solubility and dissolution behavior when using phosphate- vs. bicarbonate-buffered biorelevant media.