Step Time Flow rate (ml/min) Mobile Phase A Mobile Phase B
0 0.00 0.300 50 50
1 1.00 0.300 90 10
2 2.00 0.300 90 10
3 4.00 0.300 50 50
4 5.00 0.300 50 50
Table 1a: Gradient elution program of chromatographic separation.