Table 1.
HPLC / MS parameters for marker substrate and marker metabolite separation and detection
| CYP Iso-enzyme |
Marker substrate | Marker metabolite | Gradient/isocratic | Injection volume [μl] |
Flow rate [ml/min] |
Column temp. [°C] |
Stop time [min] |
Detection |
|---|---|---|---|---|---|---|---|---|
| 2A6 | coumarin | umbelliferone | 0–6 min linear | 10 | 0.4 | 30 | 8 | Transition (m/z) 163.0 > 107.0 |
| 2B6 | S-mephenytoin | nirvanol | 0–4 min linear | 10 | 0.4 | 30 | 6 | Transition (m/z) 205.0 > 134.0 |
| 2C8 | paclitaxel | 6α-hydroxypaclitaxel | 0–6 min linear | 10 | 0.4 | 30 | 7.5 | Transition (m/z) 870.2 > 524.9 |
| 2C9 | diclofenac | 4′-hydroxydiclofenac | 0–6 min linear | 10 | 0.4 | 30 | 7 | SIR (m/z) 265.9 / 249.9 |
| 2C19 | S-mephenytoin | 4′-hydroxymephenytoin | 0–6 min linear | 10 | 0.4 | 30 | 6 | Transition (m/z) 235.0 > 150.2 |
| 2D6 | bufuralol | hydroxybufuralol | 0–6 min linear | 10 | 0.4 | 30 | 8 | MRM (m/z) 278.2 > 186.1 |
| 2E1 | chlorzoxazone | 6-hydroxychlorzoxazone | Isocratic | 20 | 0.5 | 30 | 8 | Absorption at 298 nm |
| 3A4 | testosterone | 6β-hydroxytestosterone | 0–6 min linear | 10 | 0.4 | 30 | 7.5 | Transition (m/z) 305.0 > 287.0 |
m/z mass-to-charge ratio, SIR selected ion recording, MRM multiple reaction monitoring