Analysis of the crystallization process of a biopharmaceutical compound in the presence of impurities using process analytical technology (PAT) tools

Abstract

BACKGROUND. The crystallization of biopharmaceuticals can be problematic since, because the biosynthesis of these compounds is very difficult to control, they can contain a significant amount of impurities that have to be eliminated. In fact, impurities can lead to changes in the properties of the drug that can significantly reduce its effectiveness or even put the user in danger. The aim of this work is to exploit process analytical technology (PAT) tools to study the crystallization step of vitamin B12 crude product extracted via fermentation. RESULTS. Linear cooling crystallization experiments were performed using different conditions. The effects of solvent, cooling rate, seeding and purity of the initial material on the final size distribution and purity of the crystals were investigated through the use of UV/Vis spectroscopy, focused beam reflectance measurement (FBRM) and the CryPRINS software (Crystallization Process Informatics System). It was found that impurities strongly inhibit the growth of vitamin B12 crystals, promoting nucleation and leading to poor final crystal size distribution. CONCLUSION. Slow cooling can help to increase the purity of the final product but also generates a broad crystal size distribution because of secondary nucleation. Preparing the solution with material already crystallized once and using purified seeds helped to obtain a narrower crystal size distribution and also to reduce breakage.