UHPLC/UV/MS Operating Conditions
| Parameter | Value | ||
|---|---|---|---|
| Column: | Waters Aquity UHPLC BEH C18, 3.0 mm × 100 mm, 1.7 μm | ||
| Column Temperature: | 60 °C | ||
| Column Flow Rate: | 0.7 mL/Minute | ||
| Mobile Phase A: | Water | ||
| Mobile Phase B: | Methanol | ||
| Strong Wash: | Methanol | ||
| Weak Wash: | 80/20 Water/Methanol | ||
| Seal Wash: | 90/10 Water/Methanol | ||
| Gradient Program: | Time (min) | % Mobile Phase B | Curve |
| 0 | 20 | NA | |
| 12 | 95 | 6 | |
| 15 | 95 | 6 | |
| 15.5 | 100 | 6 | |
| 20 | 100 | 6 | |
| 22 | 20 | 1 | |
| Injection Volume: | 5 μL | ||
| Injection Mode: | Partial Loop with Needle Overfill | ||
| Strong Wash Volume: | 600 μL | ||
| Weak Wash Volume: | 1800 μL | ||
| Loop Size: | 10 μL | ||
| PDA Detector: | 220–400 nm | ||
| MS Operating Conditions | ||
|---|---|---|
| Parameter | APCI+ | APCI− |
| Scan Range (Daltons): | 110–1250 | 130–1250 |
| Corona Current (μA): | 0.5–2 | 2–5 |
| Cone Voltage (V): | 20–35 | 50 |
| Extractor (V): | 3.0 | 3.0 |
| RF (V): | 0.1 | 0.1 |
| Source Temperature (°C): | 120 | 120 |
| Probe Temperature (°C): | 550 | 550 |
| Cone Gas Flow (L/h): | 50 | 50 |
| Desolvation Gas Flow (L/h): | 600 | 600 |