Time, min

Pump A (%), aqueous buffer

Pump B (%) acetonitrile

Flow rate, mLmin⁻¹

0

90

10

0.5

0.5

90

10

0.5

1

80

20

0.5

6

20

80

0.5

8.5

20

80

0.5

8.55

90

10

0.75

10.55

90

10

0.75

10.6

90

10

0.5