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.