| Time (min) |
Mobile phase A (%) |
Mobile phase B (%) |
Comment |
| 0.0 |
80.0 |
20.0 |
Isocratic hold to elute positively charged peptide and hydrophilic formulation excipients |
| 4.0 |
80.0 |
20.0 |
| 4.1 |
0.0 |
100.0 |
Gradient step to elute heterogeneous and lipophilic polysorbates in one single peak |
| 9.0 |
0.0 |
100.0 |
| 9.1 |
80.0 |
20.0 |
Column re-equilibration |
| 12.0 |
80.0 |
20.0 |
|
