Table 1.
Chromatographic parameters for fractionation.
| Parameters | Aqueous Extract | Organic Extract | ||
|---|---|---|---|---|
| Mobile phase | Mobile phase A: Water + 10 mM ammonium acetate Mobile phase B: 100% Acetonitrile |
Mobile phase A: 95:5 Water:methanol with 0.1% formic acid and 5 mM ammonium formate Mobile phase B: 65:30:5 Isopropanol:methanol:water with 0.1% formic acid and 5 mM ammonium formate |
||
| Flow rate | 1.2 mL/min | 1.2 mL/min | ||
| Injection volume | 1 mL | 0.3 mL | ||
| Thermostat autosampler | 4 °C | 4 °C | ||
| Temperature TCC | 25 °C | 25 °C | ||
| DAD | 210 and 254 nm | 210 and 254 nm | ||
| Peak width | >0.05 min | >0.05 min | ||
| Fraction collection mode | Time based | Time based | ||
| Total time | 13 min | 13 min | ||
| Column | ZORBAX SB-C18(9.4 × 50 mm, 5 µm, p/n: 846975-202) | ZORBAX SB-C18(9.4 × 50 mm, 5 µm, p/n: 846975-202) | ||
| Time slices | 0.292 min/well | 0.292 min/well | ||
| Gradient | Time (min) 0.0 1.0 8.0 8.1 10.0 10.1 12.0 |
% Solvent B 5 5 35 95 95 5 5 |
Time (min) 0.0 1.0 8.0 11.0 11.1 12.0 |
% Solvent B 60 60 100 100 60 60 |