Table 1 Mobile phase optimization using different solvents and their mixtures
Chromatographic parameters | Acceptance criteria | Resultsa | |||||
|---|---|---|---|---|---|---|---|
Mobile phase | A | B | C | D | E | F | |
Methanol: buffer | – | 65:44 | 70:29 | 75:24 | 80:19 | 85:14 | 90:9 |
Methanol: chloroform | – | 65:44 | 70:29 | 75:24 | 80:19 | 85:14 | 90:9 |
Methanol : acetonitrile | – | 65:44 | 70:29 | 75:24 | 80:19 | 85:14 | 90:9 |
2% phosphoric acid (%w/v) | – | 1.0 | 1.0 | 1.0 | 1.0 | 1.0 | 1.0 |
Average retention time | – | 7.8 | 7.2 | 6.7 | 6.2 | 5.9 | 5.5 |
Average tailing factor | 0.8–1.5 | 1.6 | 1.36 | 1.24 | 1.13 | 1.02 | 1.23 |
Average number of theoretical plates | More than 2000 | 13,624 | 12,598 | 11,234 | 14,321 | 13,925 | 13,268 |