Table 3.
Separation parameters of HPLC/FLD method.
| Chromatographic Conditions | |
|---|---|
| Parameter | Value |
| Analytical column | Agilent ZORBAX Eclipse PAH 4.6 × 150 mm, 3.5 µm |
| Column temperature | 25 °C |
| Gradient | (A) Water, (B) Acetonitrile |
| 0 min, 40% B at 1.3 mL/min | |
| 5 min, 40% B | |
| 20 min, 100% B | |
| 30 min, 100% B | |
| 30.1 min, 40% B | |
| Post time 6 min | |
| Injection volume | 5.0 µL |
| Autosampler temperature | 18 °C |
| Diode array detector | 230 nm, band width 4 nm, reference 400 nm, reference band width 100 nm, 10 Hz |
| Fluorescence detector | Multisignal acquisition, set at λex = 260 nm and λem = 350 nm (FLD A), 330 nm (FLD B), 440 nm (FLD C), 500 nm (FLD D), 18.51 Hz |