HPLC
How to program a gradient
When using gradient with Shimadzu HPLC set-up a gradient method should be created.
The principle is easy. You state which concentration of solvent B/C/D you want at a given time point (A =100% -B - C- D) and the system will move there linearly from the previous time point. Hence for a plateau, you need to put the same concentration twice and for a steep step you need to choose two time points that are as close to each other as possible.
A=100%-B-C-D always (it is an equation), i.e. in Pump tab: B=C=D=0 and A=100% by default.
Ex. If B=20% A goes automatically to 80%.