Thus, the higher the level of organic solvent, the greater the retention of polar solutes. Flush with another 20 ml water, then condition with 30-40 ml of the mobile phase before injecting samples. Filter samples and mobile phases before use. Alternatively, filter stock buffer solutions used to prepare mobile phases, and use filtered water and HPLC-grade solvents. Column maybe stored in either water or mobile phase when not in use.
Gradient elution is possible, using decreasing concentrations of acetonitrile.
We recommend 10 mM TEAP, pH 4.4. This may not be necessary for large sialidated oligosaccharides. Otherwise, pure solutes may elute in multiple peaks or elute in the void volume.