| Time (min) | Flow rate (mL min–1) | % water (0.1% TFA) | % acetonitrile (0.1% TFA) |
| 0 | 6 | 90 | 10 |
| 25 | 6 | 60 | 40 |
| 35 | 6 | 50 | 50 |
| 40 | 6 | 30 | 70 |
| 44 | 6 | 90 | 10 |
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