2.2.3. Semi-preparative HPLC separation of sesamin andsesamolinSemi-preparative HPLC was performed to separate sesamin andsesamolin from the primary lignan crystals into two separate frac-tions. The same machine described for analytical HPLC above butfitted with a semi preparative reverse phase Phenominex Luna C18column of dimension 250 mm × 10 mm i.d., 10 m particle sizewas used. The mobile phase described for analytical HPLC abovewas run with a flow rate of 3.1 ml/min (Amarowicz et al., 2001).300 l of 1 mg/ml solution of the lignan prepared in HPLC grademethanol was injected into the HPLC. The peaks were detected at290 nm. As soon as the sesamin peak appeared at 24.34 min on themonitor of the computer, the fractions were collected manually inseparate reagent bottles repeatedly with 50 injections. Sesamolinfractions were similarly collected with the appearance of the peakat 31.75 min. Solvent from the collected fractions was evaporatedusing rotary evaporator to obtain powdered fractions of sesaminand sesamolin in two separate fractions (Fraction I and Fraction II).