Table 2007.05D.
Chromatographic conditions for LC/MS triple quad
| HPLC column | Zorbax SB-C18, 5 μm, 2.1 mm id × 50 mm, Agilent | |||
|---|---|---|---|---|
| Mobile phase A | 5.0% methanol, 0.1% formic acid, and 10 mM ammonium acetate in purified water | |||
| Mobile phase B | Acetonitrile-methanol (1 + 1 ) containing 0.1% (v/v) formic acid and 10 mM ammonium acetate in purified water | |||
| Pump program | ||||
| Time, min |
%A |
%B |
Gradient curve |
|
| 0 | 70 | 30 | NAa | |
| 13 | 30 | 70 | 1b | |
| 15 | 0 | 100 | 1 | |
| 16 | 70 | 30 | 1 | |
| 20 | 70 | 30 | NA | |
| Column temperature, °C | 40 | |||
| Flow rate, mL/min | 0.2 | |||
| Injection volume, μL | 25 | |||
| Sample introduction |
Electrospray positive (ES+) |
|
|
|
| Mode | ||||
| Mass range | Daughter scan for specific ion, dwell = 0.25 s | |||
| Source temperature, °C | 150 | |||
| Desolvation temperature, °C | 350 | |||
| Desolvation gas flow, L/h | 600 | |||
| Cone gas flow, L/h |
60 |
|
|
|
| Ions to be monitored for LC/MS confirmation | ||||
| Compound |
Parent ion (m/z) |
Daughter ions (m/z) |
Cone, V |
Collision, eV |
| [Aristolochic acid I + NH4]+ | 359.1 | 265, 281, 296 | 18 | 40, 40, 30 |
a
NA = Not applicable.
b
A gradient curve of 1 indicates a linear transition.