The gradient programme was initiated with 5% vol. of eluent A and 95% vol. of eluent B (gradient 1) and it was allowed to travel up to 30 min.
The 50% vol.:50% vol. of eluent A and eluent B (gradient 5) was achieved with a linear ramp in both
solvents and flow rate within 15 min. The gradient was reverted to original conditions within next 5 min.
The baseline was allowed to stabilize for 25 min before injecting the actual samples.
Separation of active components was achieved successfully within a total run time of 90 min