| Validation Parameters |
MP |
PP |
LRD |
| Specificity |
Specific |
Specific |
Specific |
| System Suitability |
|
|
|
| Retention time (min.) |
2.4 |
3.6 |
8.7 |
| Resolution factor (Rs) |
--- |
7.665 |
34.13 |
| No. of theoretical plates (N) |
5075 |
7067 |
37695 |
| Tailing factor (Asymmetry) |
1.0693 |
0.9955 |
0.9298 |
| Precision (Repeatability) %RSD |
|
|
|
| System (n=6) |
0.479 |
0.488 |
0.490 |
| Method (n=6) |
0.763 |
0.718 |
0.756 |
| Reproducibility |
|
|
|
| Intra-day (n=6) |
1.304 |
1.204 |
1.258 |
| Inter-day (n=6) |
1.221 |
1.324 |
1.098 |
| Accuracy (% Recovery mean ± SD) |
|
|
|
| Spike level I (n=3) |
101.87 ± 0.097 |
101.64 ± 0.19 |
101.36 ± 0.309 |
| Spike level II (n=3) |
101.57 ± 0.217 |
101.44 ± 0.269 |
101.70 ± 0.256 |
| Spike level III (n=3) |
101.93 ± 0.049 |
101.60 ± 0.310 |
101.31 ± 0.471 |
| Linearity and Range (µg/mL) |
|
|
|
| Response range |
120-180 |
12-18 |
80-120 |
| r2 |
0.9998 |
0.9999 |
0.9999 |
| Y-Intercept |
-257647 |
-8780.7 |
-25617 |
| Slope |
102422 |
91165 |
49078 |
| Sum of residual squares |
3.6E+009 |
9004199 |
1.5E+008 |
| Limit of detection/quantitation |
|
|
|
| LOD (µg/mL) |
7.15 |
0.06 |
0.46 |
| LOQ (µg/mL) |
21.68 |
0.19 |
1.40 |
| Robustness (% assay mean ± SD) |
|
|
|
| Change in flow rate |
|
|
|
| 0.8 mL/min (n=3) |
100.72 ± 1.147 |
100.50 ± 1.003 |
100.43 ± 0.806 |
| 1.2 mL/min (n=3) |
100.98 ± 0.911 |
100.61 ± 0.857 |
100.29 ± 0.734 |
| Change in detection wavelength |
|
|
|
| 261 nm (n=3) |
100.69 ± 1.195 |
100.44 ± 1.061 |
100.04 ± 0.994 |
| 265 nm (n=3) |
100.81 ± 1.047 |
100.55 ± 0.935 |
100.35 ± 0.677 |
| Change of column |
|
|
|
| Merck (n=3) |
100.83 ± 1.032 |
100.58 ± 0.900 |
100.59 ± 0.694 |
| Teknokroma (n=3) |
100.72 ± 1.177 |
100.43 ± 1.085 |
100.66 ± 0.625 |
| Change in column temperature |
|
|
|
| 23°C (n=3) |
100.87 ± 0.972 |
100.63 ± 0.846 |
100.24 ± 0.776 |
| 27°C (n=3) |
100.85 ± 0.989 |
100.51 ± 0.991 |
100.20 ± 0.814 |
