Purification of any bioactive compound obtained by biotransformation process is a critical aspect for its quality and recovery leading to its commercial viability. In this study, several solvents and their interactions were studied for the recovery of 3-demethylated colchicine (3-DMC; anti-cancer drug) from the fermentation medium. Due to inherent toxicity of chloroform, we also aimed at its removal from the solvent extraction system. A quadratic mixture model was developed using RSM with interaction terms as shown by contour/3D plots. A set of 15 experiments were performed and validated with different solvent combinations. In this study, the developed polynomial model showed a high F-value (362.36) and R-2 value (0.99) which suggests the suitability of D-optimal design to predict the responses accurately. The results indicated higher recoveries of 5.95 g/l of 3-DMC, without the use of chloroform enabling efficient extraction of 3-DMC for commercial applications. (C) 2015 Elsevier Ltd. All rights reserved.