Sephadex g 15 column

Yellow wastewater from tofu production was condensed (1/5 of its original volume) and precipitated using three times the volume of cold ethanol (4 °C). The supernatant was then freeze-dried and termed crude SBOS. The crude SBOS isolated from wastewater of tofu production were redissolved in distilled water (30 mg/mL), and 2 mL solution was passed through the Sephadex G-15 column (1.0 × 70 cm, Sigma, Shanghai, China) at a flow rate of 0.5 mL/min with ultra-pure water. The SBOS were pooled based on the elution curve monitored by the phenol–sulfuric acid method [15 (link)].

The roots of Panax ginseng were obtained from a local market (Changchun, China), and authenticated by Prof. Jiang (Jilin Academy of Chinese Medicine and Material Medical Science, Changchun, China). The roots (1 kg) were first pulverized by grinder and reflux extracted with 85% ethanol to remove the saponine. The residue was decocted with 5 L water three times, and then the rinsing fluids were merged into one portion, and evaporated to 200 mL. Subsequently, the aqueous extract was forced through a cellulose superfiltration system (molecular weight cut-off is 20,000 Da) to give two different molecular weight fractions. The low molecular weight fraction was dialyzed against water by using a dialysis sac with a 1000 Da cut-off. The contents out of the dialysis sac were collected, concentrated, lyophilized and loaded on a Sephadex G-15 column (Ф2.5 × 95 cm, from Sigma Chemical Co., Saint Louis, MO, USA), which eluted with pH 7.0 phosphate-buffered saline at a flow rate of 0.2 mL/min, to obtain purified glycopeptide. The eluent was collected by a fraction collector, and named Gg, and then purification of the glycopeptides was carried out by HPLC. The two major portions were collected for further characterizing.

Doxorubicin was purchased from Sigma-Aldrich (St. Louis, MO). Dried heartwood powder of Caesalpinia sappan L. was obtained from B2P2TOOT (Tawangmangu, Indonesia). Dried powder was extracted in methanol by maceration to get the methanol extract. The methanol extract was diluted as 4:1 methanol/water and then partitioned with hexane. The aqueous layer was fractioned with ethyl acetate and concentrated with a vacuum rotary evaporator to get the ethyl acetate fraction. Brazilin (0.245 g) (Figure 1) was obtained by separation of ethyl acetate fractions using Sephadex G-15 column (Sigma-Aldrich) chromatography (15 × 7 cm) with gradient polarity of the mobile phase (CHCl₃:MeOH) and was collected using thin-layer chromatography.