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HPLC Question: Consider the following system: Column 2.1mm i.d., Flow Rate 0.3ml/min.?

Mobile phase gradient Time (min)
0Min A 98% 2%B
20min B 0% 100B

Just dial it into the HPLC controller.

The total rate is constant at 0.3ml/min, this is made up of flow A and flow B. At t=0 A is pumping 0.3*0.98 ml/min and B at 0.3*0.02ml/min. After 20 min A is pumping 0.3*0 ml/min and B 0.3*1 ml/min. To calculate the intermediate points plot these flows out, draw a straight line from A t=0 to A t=20 and read off the flow at the required time. At t=10min flow A = 0.3*0.49 ml/min and B= 0.3*0.51 ml/min

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