Optimization of chromatographic conditions for reversed-phase (RP)-HPLC method
| Chromatographic parameters | Variations | Peak shape |
|---|---|---|
| Mobile phase composition (v/v) | ||
| ACN:Methanol:Water | 50:30:20 | No ideal peak |
| ACN:Water | 50:50 | Broad peak |
| 60:40 | Not symmetric | |
| 70:30 | Sharp resolved peak, tailing | |
| 80:20 | Well resolved peak | |
| pH variation | ||
| ACN:Orthophosphoric acid (pH 4.0); | 4.0 | Broad peaks |
| optimized ratio (80:20) | 3.0 | Resolved sharp peak, tailing |
| 3.5 | Sharp, well separated peaks |