Table 1 Optimization mobile phase and column in method development
Trail no. | Mobile phase composition | Column | Observation | Inference |
|---|---|---|---|---|
1 | Monobasic sodium phosphate: acetonitrile (60:40 v/v) | ACQUITY UPLC HSST3 100 mm × 1 mm 1.8 μm | Delta (6,7) fulvestrant impurity not separated from fulvestrant | Method rejected |
2 | 0.1% v/v TEA: acetonitrile (60:40 v/v) | ACQUITY UPLC HSST3 100 mm × 1 mm 1.8 μm | Impurities are not separated from fulvestrant peak | Method rejected |
3 | 0.1% v/v OPA: methanol (600:400 v/v/v) | ACQUITY UPLC Phenyl 150 mm × 2.1 mm 1.7 μm | Impurities are separated from fulvestrant. But sulfone impurity was eluted very late | Method rejected |
4 | 0.1% v/v OPA: acetonitrile: methanol (300:400:300 v/v/v) | ACQUITY UPLC Phenyl 150 mm × 2.1 mm 1.7 μm | Impurities are not well separated from fulvestrant | Method rejected |
5 | 0.1% v/v OPA: acetonitrile and methanol (300;400;300 v/v/v) | BEH shield RP18, 50 mm × 2.1 mm 1.7 μm | Impurities are well separated from fulvestrant with good peak shape and less run time | Approved |