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| Fractions | Starting %B | Ending %B |
|---|---|---|
| 1 and 2 | 2% | 19% |
| 3 and 4 | 4% | 24% |
| 5 and 6 | 7% | 27% |
| 7 and 8 | 10% | 28% |
| 9 and 10 | 13% | 35% |
The above gradients are each 72 minutes in duration, after which there is a step up to 70% B, followed by a return to starting conditions for column equilibration. The total run time of each method is 90 minutes.
