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A: 0.1% TFA in Water B: 90% ACN 0.09% TFA in Water
Gradient Profile:
Step No.
Time(min)
%A
%B
1
0
80
20
2
25
45
55
Flow Rate:
1 mL/min
Temperature:
45°C
Detection:
UV-Vis Abs.-Variable Wave.(UV) @ 214 nm (22°C)
Sample Note:
Reversed phase runs of PEGylated ribonuclease A using different gradient conditions on a Jupiter 300 C4 column (gradient length 25 min). Note that if initial %B is increased much above 25% then retention and resolution of PEGylated forms is lost. Gradient slopes of between 1-2%/min appear to be a good compromise between resolution and run time.