Spherisorb s10 ods2 column

Dieckol was a light brown powder and was provided by Botamedi, Inc. (Jeju, Korea); its specifications are available elsewhere [25 (link)]. Briefly, the whole E. cava plant was collected off the coast of Jeju Island, South Korea. Dried E. cava powder was extracted with 70% aqueous ethanol and partitioned between water and ethyl acetate. The ethyl acetate fraction was subjected to octadecylsilyl (ODS) column chromatography followed by gel filtration in a Sephadex LH-20 column equilibrated with methanol. Final purification was accomplished by HPLC (Waters Spherisorb S10 ODS2 column (20 × 250 mm); eluent, 30% methanol; flow rate, 3.5 mL/min) to isolate dieckol (98.5 wt%). As the vehicle, 2 wt%, 5000–10,000 kDa high-molecular-weight HA was used (MNH Bio, Hwaseong, Korea). The dieckol+vehicle was prepared as follows: 0.2 g of dieckol was mixed with 3 mL of 10% ethanol and 0.9 mL of normal saline at 37 °C and centrifuged for 30 min. Next, 16.1 mL of high-molecular-weight HA were stirred into the solution for 24 h.

The seaweed (1000 g) was extracted with 95% ethanol (10 L) for 3 h in a water bath at 50°C. The solvent was evaporated in vacuo to give a gummy extract, which was partitioned between ethyl acetate and water. The ethyl acetate soluble portion was subjected to ODS (Octadecylsilane) column chromatography followed by gel filtration on Sephadex LH-20 with methanol. Final purification of individual compound was accomplished by HPLC (Waters Spherisorb S10 ODS2 column (20 × 250mm); eluent, 30% MeOH; flow rate, 3.5mL/min). By means of 1H-NMR spectra and comparison with published data, phlorofurofukoeckol-A (PFF-A, C5) were identified. Phlorofurofukoeckol-A: white solid; UV (EtOH) λmax 227 nm; 1H-NMR (DMSO-d6, 500MHz) δ 5.73 (d. J = 1.85, 1H), 5.76 (d, J = 1.85, 1H), 5.83 (t, J = 1.85, 1H), 5.84 (t, J = 1.85, 1H), 6.29 (s, 1H), 6.43 (s, 1H), 6.72 (s, 1H), 8.22 (s, OH), 9.19 (s, OH), 9.22 (s, OH), 9.45 (s, OH), 9.86 (s, OH), 9.88 (s, OH), 10.15 (s, OH); 13C-NMR (DMSO-d6, 125MHz) δ 94.3, 94.0, 95.3, 96.9, 97.0, 98.8, 99.6, 103.8, 103.9, 120.6, 123.0, 123.1, 126.9, 134.5, 137.4, 142.5, 145.3, 147.0, 147.5, 150.0, 150.9, 151.4, 159.4, 159.5, 160.5, 160.8. FABMS m/z 603.1 (M+H)+.