Comparative Study Of Endotoxin Removal From Recombinant Activepharmaceutical Ingredient

Endotoxins include lipopolysaccharide(LPS), found in the outer membrane of Gram negative bacteria, is the major cause of pyrogenic reactions that can cause side effects such as endotoxin shock, tissue injury and even death. In aqueous solutions, endotoxins can exist in various states of aggregation that only exerts its toxic effect when in freeform. Their peculiar structure, and their broad spectrum of biological activities has resulted in worldwide research to find the suitable method to reduce this contamination from parenteral.The ultrafiltration and ion exchange chromatography permit the purification and disinfection of solutions. These techniques are physical solution treatmentprocedures that can be used to remove these pyrogenic molecules from Active Pharmaceutical Ingredient(API) with different concentration of Sodium Deoxycholate and Tween 20 as a chemical method.Response Surface Methodology(RSM), is a statistical technique, itsapplicationis to optimize a response (output variable) which is influenced by several independent variables (input variables). RSM software with 2 parameters calculated results of Ion Exchange Chromatography with different concentration of Sodium Deoxycholate and Tween.The Sodium deoxycholatewith 1/5% concentration for 24 h incubation in ultrafilration method was the most effective concentration in reducing the amount of endotoxin. Although in ion exchange chromatography, Sodium deoxycholatewith 1/5% concentration and 0/75% tween had the best result in endotoxin reducing, but increasing in tween concentration wasn’t improved purification, reversely Sodium deoxycholate.The results showed that ion exchange chromatography was more efficient with a low concentration of endotoxin (98% of LPS removed) compared to ultrafiltration.

Keywords:

Lipopolysaccharides , ion exchange chromatography , Ultrafiltration , Sodium Deoxycholate , Endotoxins , RSM