Table 1.
Step1 | Start | Sec2 | Loading pump Flow (mL/min) | Grad | %A | %B | %C | %D | Tee3 | Loop4 | Eluting pump Flow (mL/min) | Grad | %A | %B |
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
1 | 0.00 | 20 | 3.00 | Step | 100 | - | - | - | - | Out | 0.37 | Step | 98 | 2.0 |
2 | 0.33 | 120 | 0.07 | Step | 100 | - | - | - | T | In | 0.3 | Step | 98 | 2.0 |
3 | 2.33 | 60 | 2.00 | Step | - | 100 | - | - | - | In | 0.36 | Ramp | 65 | 35 |
4 | 3.33 | 45 | 2.00 | Step | - | 100 | - | - | - | Out | 0.37 | Step | 65 | 35 |
5 | 4.08 | 20 | 2.00 | Step | - | - | 80 | 20 | - | In | 0.38 | Ramp | 25 | 75 |
6 | 4.42 | 25 | 2.00 | Step | - | - | 80 | 20 | - | Out | 0.39 | Step | 2.0 | 98 |
7 | 4.83 | 25 | 2.00 | Step | - | - | 80 | 20 | - | In | 0.38 | Step | 98 | 2.0 |
8 | 5.25 | 90 | 2.00 | Step | 100 | - | - | - | - | Out | 0.37 | Step | 98 | 2.0 |
At each step, the mobile phases for both loading and eluting pump were introduced by step gradient
time as second
the loop connected to analytical column
the transfer loop connected to HT column