Chromatography
Chromatography was carried out at ambient temperature.
The mobile phase consisted of acetonitrile and water
(90:10). The flow-rate of the mobile phase was 1mL/min.
10 μL of each of the standard solutions of the four
compounds were injected in the HPLC to prepare a
calibration graph. Then ten micro liters of sample solution
was injected and concentration of each component
was determined through the calibration graphs of the
respective standards.