Column

Inertsil ODS-3, 150 × 4.6 mm 3 µ

Column temperature

35˚C

Wave length

270 nm for Imp-D and 340 nm for remaining impurities

Injection volume

100 µL

Run time

70 minutes

Gradient

90% A upto 8 min (isocratic) with 1.0 mL

Linear gradient to 75% A at 20 min with 1.0 mL

Linear gradient to 70% A at 45 min with 1.0 mL

Linear gradient to 20% A at 50 min with 1.2 mL

Linear gradient to 0% A at 52 min with 1.2 mL

0% A from 52 min to 60 min with 1.2 mL

Linear gradient to 90% A at 62 min with 1.0 mL

90% A from 62 min to 70 min with 1.0 mL