Parameters adjustments, tested values and results obtained for robustness
| Parameters (Rs>3.5) | Authorised adjustments | Tested values | System suitability (R>3.5) | Nystatin A1 retention time (min) |
|---|---|---|---|---|
| Ammonium acetate buffer 0.05 M pH = 6.0, methanol 35/65 (v/v) | Proportion of ammonium acetate buffer: ±30% relative | 25:75 (v/v) | 1.8 | No separation |
| 30:70 (v/v) | 1.9 | Incomplete separation | ||
| 35:65 (v/v) | 5.4 | 8.1 | ||
| 40:60 (v/v) | 7.3 | 15.7 | ||
| 45:55 (v/v) | 8.3 | 37.5 | ||
| pH buffer: 6.0 | ±0.2 | 5.8 | 6.3 | 8.0 |
| 6.0 | 5.4 | 8.1 | ||
| 6.2 | 5.9 | 8.1 | ||
| Buffer molarity: 0.05 M | ±10% | 0.045 M | 5.8 | 8.1 |
| 0.05 M | 5.4 | 8.1 | ||
| 0.055 M | 6.2 | 8.1 | ||
| Flow rate: 1.0 mL/min | ±50% | 0.5 mL | 7.4 | 16.9 |
| 1.0 mL | 5.4 | 8.1 | ||
| 1.5 mL | 4.9 | 5.5 | ||
| Column temperature: 30°C | ±10°C | 20°C | 6.4 | 10.2 |
| 30°C | 5.4 | 8.1 | ||
| 40°C | 6.0 | 6.8 |
The values corresponding to the described method are in bold.
The values out of specifications are in italic.