9. Appendix

9.1 Sep-Pak filtration for carbohydrates (courtesy Dr. Gordon Alton)

For oligosaccharides with hydrophobic aglycons the following conditions are recommended for C18 Sep-Pak cartridges:

1. Wash with 5 mL of methanol then 10 mL of water
2. Load oligosaccharide in water or buffer, the more dilute the better, use a flow rate of less than 10 mL/min (slower is better)
3. Wash cartridge with 10 mL of water (flow rate is not important),
   keep all solutions until you know where the carbohydrate is!
4. Elute oligosaccharide with 5 or 10 mL of methanol.

One C18 Sep-Pak can hold up to 10 mg of an octyl glycoside trisaccharide, but is different for each sugar, aglycon, flow rate and load buffer situation. Determine experimentally what works best for your compound.