Preparative separation of sesamin and sesamolin from defatted sesame meal via centrifugal partition chromatography with consecutive sample injection

• Defatted sesame meal is a good source to isolate and purify sesamin and sesamolin.
• Consecutive sample injection into CPC was developed to purify sesamin and sesamolin from defatted sesame meal extract.
• Scale-up of CPC with 1000 mL rotor was carried out according to 100 mL rotor results.
• The antioxidant response element (ARE) activating effects of sesamin and sesamolin were evaluated by luciferase assays in HepG2 cells.

A preparative separation method using consecutive sample injection centrifugal partition chromatography (CPC) was developed to obtain sesamin and sesamolin from defatted sesame meal extracts. A two-phase solvent system consisting of n-hexane–ethyl acetate–methanol–water (8:2:8:2, v/v) was applied in reversed-phase mode (descending mode). Preliminary experiments with an SCPC-100 (column volume: 100 mL) were performed to select the appropriate two-phase solvent system and sample injection times; these parameters were then used with an SCPC-1000 (column volume: 1000 mL) in a 10-fold scale-up preparative run. A sample containing 3 g of crude extract was consecutively injected four times onto the SCPC-1000, which yielded 328 mg of sesamin and 168 mg of sesamolin. These compounds were analyzed by high-performance liquid chromatography and determined to have purities of 95.6% and 93.9%, respectively. Sesamin and sesamolin (30 μM) increased antioxidant response element (ARE) luciferase activity 2.6-fold and 1.9-fold, respectively.