LC analysis was performed on an Agilent 6460 triple Quad LC/MS system with water (A) and acetonitrile (B), both with 0.1% formic acid as mobile phase (all chemicals from Burdick and Jackson (Honeywell)). Separation was on an Agilent Poroshell HPH-C8, 2.7 μm, 3.0 × 50 mm column at a flow rate of 0.5 mL/min with a 10 μL injection volume. Elution started at 20% B (organic) for 0–5 mins, 5–15 mins to 90% B and 15–20 mins down to 20% (B). Column eluent passed into an Agilent 6460 triple quad LC/MS with a jet stream electrospray ionization source operated in a positive mode. Source and acquisition parameters are presented in Table 1.
Agilent 6460 triple Quadrupole LC/MS parameters for the analysis of antiretroviral compounds with positive electrospray ionization mode
| Parameter . | Value . |
|---|---|
| Gas Temperature | 300 °C |
| Drying Gas | 5.1 L/min |
| Nebuliser | 45 psi |
| Sheath Gas Temperature | 250 °C |
| Sheath Gas Flow | 11 L/min |
| Nozzle Voltage | 300 V |
| Fragmentor | 135 V |
| Acquisition range | 50–1500 m/z |
| Acquisition rate | 0.33 cycles/s |
| Parameter . | Value . |
|---|---|
| Gas Temperature | 300 °C |
| Drying Gas | 5.1 L/min |
| Nebuliser | 45 psi |
| Sheath Gas Temperature | 250 °C |
| Sheath Gas Flow | 11 L/min |
| Nozzle Voltage | 300 V |
| Fragmentor | 135 V |
| Acquisition range | 50–1500 m/z |
| Acquisition rate | 0.33 cycles/s |