Table 2 Showing optimized chromatographic conditions
Parameter | Condition | ||
|---|---|---|---|
HPLC instrument | Agilent 1260 Infinity II | ||
Column | Agilent Zorbax RX—C18 (150 × 4.6 mm, 5 micron), 80°A | ||
Wavelength | 254 nm | ||
Mobile phase | Mobile phase A: 85% Mobile phase B: 15% | ||
Gradient program | Time minutes | Mobile phase A | Mobile phase B |
0 | 85 | 15 | |
12.5 | 50 | 50 | |
15.5 | 50 | 50 | |
17.5 | 85 | 15 | |
20.0 | 85 | 15 | |
Diluent | ACN | ||
Retention time | 8.43 min | ||
Run time | 20 min | ||
Injection volume | 10 µL | ||
Column oven temperature | 30 0C | ||
Flow rate | 1.2 mL/min | ||
Column temperature | 35 °C (± 2 °C allowed by Robustness) | ||