Statistically designed enzymatic hydrolysis of an icariin/β-cyclodextrin inclusion complex optimized for production of icaritin

Abstract

This study focuses on the preparation and enzymic hydrolysis of an icariin/β-cyclodextrin inclusion complex to efficiently generate icaritin. The physical characteristics of the inclusion complex were evaluated by differential scanning calorimetry (DSC). Enzymatic hydrolysis was optimized for the conversion of icariin/β-cyclodextrin complex to icaritin by Box–Behnken statistical design. The inclusion complex formulation increased the solubility of icariin approximately 17-fold, from 29.2 to 513.5 μg/mL at 60 °C. The optimum conditions were predicted by Box–Behnken statistical design as follows: 60 °C, pH 7.0, the ratio of enzyme/substrate (1:1.1) and reaction time 7 h. Under the optimal conditions the conversion of icariin was 97.91% and the reaction time was decreased by 68% compared with that without β-CD inclusion. Product analysis by melting point, ESI-MS, UV, IR, 1H NMR and 13C NMR confirmed the authenticity of icaritin with a purity of 99.3% and a yield of 473 mg of icaritin from 1.1 g icariin.

Keywords

• Icariin/β-CD inclusion complex
• Snailase
• Enzymatic hydrolysis
• Box–Behnken statistical design
• Icaritin