| Time | Solvent A (Acetonitrile) | Solvent B (Water) | Flow Rate (mL/min) |
| 0 | 5% | 95% | Flow rate depends on the column packing and its size. |
| 2 | 5% | 95% | |
| 35 | 95% | 5% | |
| 38 | 100% | 0% | |
| 40 | 5% | 95% |
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