Table 1.
Valve and pumps overlay timing chart.a
| Valve | SPE pump (Binary Pump #1) | Analytical pump (Binary Pump #2) | ||||||
|---|---|---|---|---|---|---|---|---|
| Time (min) | Step | Valve Pos. | Time (min) | Methanol (%) | Flow Rate (mL min−1) | Time (min) | Acetonitrile (%) | Flow Rate (mL min−1) |
| 0 | 1 | 1 | 0 | 10 | 1.00 | 0 | 30.00 | 1.00 |
| 1.00 | 10 | 1.00 | ||||||
| 1.10 | 10 | 4.00 | ||||||
| 3.50 | 2 | 2 | 3.50 | 10 | 4.00 | |||
| 3.60 | 50 | 0 | ||||||
| 4.50 | 3 | 1 | ||||||
| 4.60 | 50 | 0 | ||||||
| 5.00 | 100 | 4.00 | ||||||
| 8.20 | 100.00 | 1.00 | ||||||
| 10.00 | 100 | 4.00 | 10.00 | 100.00 | 1.00 | |||
| 10.10 | 10 | 1.00 | ||||||
| 10.50 | 30.00 | 1.00 | ||||||
| 12.00 | 12.00 | 10 | 1.00 | 12.00 | 30.00 | 1.00 | ||
a
The aqueous mobile phase for the SPE and HPLC separation was 0.1% acetic acid in water.
The organic mobile phase was methanol (SPE) and acetonitrile (HPLC).