Analysis was performed on a Waters platform, composed of a Waters 2695 HPLC unit equipped with a Waters 2998 photodiode array (PDA) detector and a Luna silica column (4.6 mm i.d. ×250 mm, 5 µm particle size; Phenomenex, Torrance, CA, USA). The unit was coupled with a Bruker Esquire 3000 Plus ion trap mass spectrometer (Bruker–Franzen Analytik GmbH, Bremen, Germany) equipped with ESI. The PCBLs was dissolved in methanol to prepare a 20 mg/mL solution. The mobile phases consisted of hexane/methanol/ethyl acetate (10∶3∶1, v/v/v) (A) and hexane/methanol/ethyl acetate (1∶3∶1, v/v/v) (B). Separations were done by linear gradient at 37°C at 1 mL/min flow rate as follows: 0–90 min, 83.6% A; 90–130 min, 83.6–19.4% A; 130–150 min, 19.4% A. The PDA detector was set to 280 nm and scanning was done from 200 to 400 nm.
Preparation was performed on a Phenomenex (Torrance, CA, USA) Luna silica preparative column (21.2 mm i.d. ×250 mm) with a 5 µm particle size at 37°C. A Shimadzu system equipped with a CBM-20A module, an SIL-10AP autosampler, an SPD-20A UV–vis detector, and two LC-8A pumps was used. Elution was the same as the method described above. The flow rate was 21.6 mL/min and the absorption wavelength was at 280 nm. On a given run, 1 mL (200 mg/mL) extract was applied. The fractions were collected and evaporated under vacuum.
Preparation was performed on a Phenomenex (Torrance, CA, USA) Luna silica preparative column (21.2 mm i.d. ×250 mm) with a 5 µm particle size at 37°C. A Shimadzu system equipped with a CBM-20A module, an SIL-10AP autosampler, an SPD-20A UV–vis detector, and two LC-8A pumps was used. Elution was the same as the method described above. The flow rate was 21.6 mL/min and the absorption wavelength was at 280 nm. On a given run, 1 mL (200 mg/mL) extract was applied. The fractions were collected and evaporated under vacuum.