Table 2. Optimization of the HPLC Method for the Bioanalysis of 5-FU Using CCD.
| factors
and their levels |
responses | goal | ||
|---|---|---|---|---|
| design run | A | B | ||
| 1 | 0.0 | –1.0 | peak retention time (min) | <5 |
| 2 | –1.0 | 1.0 | ||
| 3 | 0.0 | 0.0 | peak area (mAU) | >10,000 |
| 4 | 0.0 | 0.0 | ||
| 5 | 1.0 | 0.0 | ||
| 6 | –1.0 | –1.0 | ||
| 7 | 1.0 | –1.0 | ||
| 8 | –1.0 | 0.0 | ||
| 9 | 0.0 | 1.0 | ||
| 10 | 0.0 | 0.0 | ||
| 11 | 1.0 | 1.0 | ||
| 12 | 0.0 | 1.0 | ||
| 13 | 0.0 | 0.0 | ||
| statistical
analysis | ||||||
|---|---|---|---|---|---|---|
| evaluation parameter | low (−1) | middle (0) | high (+1) | model | p-value | R2 |
| A: mobile phase ratio | 92:8 | 94.6 | 96:4 | quadratic | 0.0023 | 0.9784** |
| B: column temperature (°C) | 30* | 35 | 40** | quadratic | 0.0019 | 0.9597* |
* and ** indicated the lower and the higher values, respectively.