| LC | Mobile phase | A: Methanol + 0.1% AA* and B: Water + 0.1% AA |
| Flow rate | 0.70 mL/min |
| Injection volume | 20 µL |
| Gradient elution | 5 min – 95% B, 20 min – 50% B, 25 min – 50% B, 30 min – 10% B, 37 min– 10% B, 39 min – 95% B |
| Column temp | 35°C |
| Running time | 39 min |
| Post time | 5 min |
| Detection (λ) | 278 nm (caffeine), 325 nm (CGA) and 268 nm (trigonelline) |
| Spectra range | 190-400 nm |
|
| MS | Ionization mode | Atmospheric pressure ionization- electrospray (API – ES) |
| Data recording | Positive ion mode |
| Screening (m/z) | 100.00-1500.00 |
| Spray chamber | Gas temp: 350oC, neb press: 40 psi, volt: 4 kv |