Analytical conditions for substrates in fresh and cryopreserved rat hepatocyte suspensions
Substrate | Column | Mobile Phase | Detection | Flow Rate | Internal Standard | Standard Curve Range |
|---|---|---|---|---|---|---|
| ml · min -1 | nmol | |||||
| TOL | Beckman Ultrasphere 5ODS 150 × 4.6 mm (5 μm) | 40% methanol: 60% 0.05% orthophosphoric acid (v/v) | UV 230 nm | 1.0 | Chlorpropamide | 0.125-12.5 |
| 7-EC | Fluorescence excitation 375 nm, emission 452 nm | 0.00125-0.25 | ||||
| PHE | Hypersil BDS C18 150 × 4.6 mm (5 μm) | 26% acetonitrile: 74% 0.05% orthophosphoric acid (v/v) | UV 230 nm | 1.0 | 0.0025-1.25 | |
| NDZ | Beckman Ultrasphere 5ODS 150 × 4.6 mm (5 μm) | 50% methanol: 50% water: 0.02% TEA (v/v/v) pH 7 | UV 236 nm | 1.0 | Diazepam | 0.005-1.25 |
| DZ | Beckman Ultrasphere 5ODS 150 × 4.6 mm (5 μm) | 57% methanol: 43% water: 0.02% TEA (v/v/v) pH 7 | UV 236 nm | 1.0 | Prazepam | 0.125-2.5 |
| DEX | Ultra Techsphere C8 150 × 4.6 mm (5 μm) | 35% acetonitrile: 65% water: 0.1% TEA (v/v), pH 6 | Fluorescence excitation 280 nm, emission 310 nm | 0.6 | Codeine | 0.05-1.25 |
| WAR | Hichrom C18 250 × 4.6 mm (5 μm) | 38% acetonitrile: 74% 0.5% orthophosphoric acid (v/v) | UV 313 nm | 1.0 | 7-Hydroxycoumarin | 0.0125-1.25 |
| PROP | Hichrom C18 250 × 4.6 mm (5 μm) | 22% acetonitrile: 22% methanol: 56% water: 1% acetic acid (v/v) | Fluorescence excitation 295 nm, emission 340 nm | 1.0 | Codeine | 0.0625-1.25 |