Paper Title
Optimization of Extraction Conditions for Determination of Androstadienedione and its Major Impurities from Fermentation Broth by Statistical Techniques

Recent pharmaceutical regulatory guidelines have stressed upon the importance of applying quality by design (QbD) principles to ensure that process leads to build in quality in the product. This research paper outlines the application of QbD principles for optimization of extraction conditions to estimate maximum activity of androstadienedione (ADD) from fermentation broth. The study is multi response study in which along with ADD activity, its two main impurities viz.; androstenedione and C22 alcohol were estimated to maximum extent which were generated during the fermentation process. Response surface methodology was employed to assess interactive and individual effects of three main independent variables on extraction efficiency of ADD and its major impurities. The independent variables studied were pH, temperature and sonication time used for preparation of samples. The predicted values of ADD activity, and its major impurities content were in agreement with experimental responses obtained when central composite design was utilized during optimization of extraction process. Optimization results displayed that maximum extraction efficiency of ADD and % of its two major impurities from fermentation broth was achieved at optimum conditions: sonication time 10 min, the pH of sample solution 7.0 and temperature of extraction 300 C, using acetone as the extraction solvent. The activity of ADD and % of both impurities were estimated by HPLC.