Efficient preparation of narcissin from Opuntia ficus-indica fruits by combination of response surface methodology and high-speed countercurrent chromatography

Background: Narcissin is well known for their various biological activities. Objective: To recover narcissin from the Opuntia ficus-indica fruits (OFIF), an efficient method was developed by a combination of response surface methodology (RSM) and high-speed countercurrent chromatography (HSCCC). Materials and Methods: Optimization of extraction conditions of narcissin from OFIF was determined using RSM with three-level-three-factor Box-Behnken design (BBD). Then, a rapid and efficient method for the isolation of narcissin from the rich narcissin extracts was developed using HSCCC. Results: Regression analysis showed a good fit of the experimental data and the optimal condition was obtained as extraction time (X₁), 6.02 h; solvent to material ratio (X₂), 8.16 mL/g; and ethanol concentration (X₃), 93.48%. Then, the rich narcissin extracts were separated by HSCCC with a two-phase solvent system composed of n-hexane: ethyl acetate: methanol: water (1.5:5:5:1.5, v/v/v/v) in one step within 60 min. As a result, 12 mg of narcissin was isolated from 100 mg of crude extract with purities of 98.5%, as determined by high-performance liquid chromatography (HPLC). Conclusion: This study can be useful to the development of industrial extraction processes, including further studies concerning the optimal number of sequential steps to enhance the efficacy of a large-scale extraction system.