Oleuropein is the main active substance in foods or functional foods produced from olive (Olea europaea L.) leaves. In the present study, the combinative technology off line of HSCCC-PHPLC based on dual wavelength was used to separate highly purified oleuropein from oleuropein extract. Response surface methodology was used to optimize the conditions of HSCCC. Furthermore, a large amount of higher purified oleuropein was obtained through HSCCC at the wavelength of 254 nm, and oleuropein with the purity greater than 98.5% was obtained by PHPLC at the wavelength of 300 nm. Finally, the purity and structure identification of highly purified oleuropein were determined by various methods and its stability was investigated. As a result, oleuropein was stable in solution, and had good stability under the condition of dark storage at 4°C within a week or under the condition of dark storage at -20°C within one year. PRACTICAL APPLICATION: In this study, an efficient method for purification and refining of oleuropein by combinative technology off line of HSCCC-PHPLC based on dual wavelength was established. Oleuropein with the purity greater than 98.5% was macro-obtained via the technology. The highly purified oleuropein could be used to control the quality of olive products.
Keywords: dual wavelength; high-speed countercurrent chromatography; highly purified oleuropein; response surface methodology.
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