OPTIMIZATION OF AMMONIUM SULFATE PRECIPITATION AND AQUEOUS TWO-PHASE SYSTEM METHOD FOR PURIFICATION OF C-PHYCOCYANIN FROM SPIRULINA PLATENSIS

• Main Author: Dutiara, Thesar
• Format: Final Project
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/86237
• Institution: Institut Teknologi Bandung

Spirulina platensis is a photosynthetic, blue-green, spiral or bulb-shaped microalgae, and is high in C-Phycocyanin (CPC) so it tends to be blue-green. Preliminary research shows that CPC has the potential to be used as a substitute substrate in the determination of pancreatin protease activity. In the pharmaceutical field, pancreatin is used as an oral agent for pancreatic enzyme-replacement therapy (PERT) for patients with pancreatic exocrine insufficiency (PEI). Therefore, the determination of pancreatin protease activity needs to be done to ensure the quality of drug raw materials. This study aims to optimize the ammonium sulfate precipitation and aqueous two-phase system methods to obtain CPC fractions with high concentration and purity. Optimization was carried out on several factors including dry biomass concentration, ammonium sulfate concentration, type of PEG, and concentration of PEG used. The isolation of CPC from Spirulina platensis was carried out through 2 stages, which are extraction and purification. Extraction was carried out using a combination method of homogenization, sonication, centrifugation, and filtration to obtain crude extract (EK). Purification was done using a combination of ammonium sulfate precipitation fractionation and aqueous two-phase system (ATPS) to obtain CPC fraction (F-CPC). The optimum conditions for CPC extraction were 5% biomass concentration, 40% ammonium sulfate, and 6% PEG 6000.