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Time program for the chromatographic separation procedure.
| Total time (min) | Flow rate (mL/min) | Mobile phase solvent A (%) | Mobile phase solvent B (%) |
|---|---|---|---|
| 0 | 0.4 | 95 | 5 |
| 1 | 0.4 | 95 | 5 |
| 4 | 0.4 | 70 | 30 |
| 7 | 0.4 | 50 | 50 |
| 13.5 | 0.5 | 5 | 95 |
| 14 | 0.5 | 5 | 95 |
| 14.5 | 0.4 | 95 | 5 |
| 16 | 0.4 | 95 | 5 |
Solvent A: 97.9% water with 2% methanol and 0.1% formic acid. Solvent B: 99.9% methanol with 0.1% formic acid.
