Table 4.
Application of method to quantification and method validation parameters for quantification of 2-O-methyl-anziaic acid and lecanoric acid from the lichen Melanelia subaurifera and Melanelia fuliginosa
| Application of method to quantification of 2-O-methyl-anziaic acid and lecanoric acid | ||
|---|---|---|
| Sample | Concentration (mg/gm)a | % CV |
| 2-O-methyl-anziaic acid | ||
| Melanelia fuliginosa | 4.24 ± 0.04 | 4.39 |
| Lecanoric acid | ||
| Melanelia subaurifera | 8.71 ± 0.01 | 3.24 |
| Method validation parameters | ANZ | LEC |
|---|---|---|
| LOD (µg/ml) | 0.71 | 0.26 |
| LOQ (µg/ml) | 2.37 | 0.87 |
| Linear range (µg/ml) | 15–1000 | 1.5–600 |
| Mean correlation coefficient (r2) | 1 | 1 |
| System suitability (% CV, n = 6) | 0.89 | 0.92 |
| Retention time (min) | 7.44 | 2.47 |
| Accuracy and precision (% CV, n = 3) | ||
| Intraday | 0.64–1.11 | 0.89–1.11 |
| Interday | 0.96–1.42 | 1.00–1.56 |
| Stability | 24 h | |
| Melanelia subaurifera | 97.18 % | |
| Melanelia fuliginosa | 94.53 % | |
| Stability | 20 days | |
| Melanelia subaurifera | 95.52 % | |
| Melanelia fuliginosa | 92.87 % | |
| Mobile phase MeOH: d/w, v/v | Retention time (min) | |
|---|---|---|
| ANZ | LEC | |
| Ruggedness | ||
| 81:19 | 7.32 | 2.38 |
| 80:20 | 7.44 | 2.47 |
| 79:21 | 7.69 | 2.54 |
| Flow rate change (ml/min) | ||
| 0.9 | 7.31 | 2.35 |
| 1.0 | 7.44 | 2.47 |
| 1.1 | 6.80 | 2.10 |
aMean ± SD, n = 3 (batches)