Optimization Of Separation:

The basic aim of the separation of components of a complex mixture is the adequate resolution along with highest purity in the shortest possible time. Though it is a common practice to follow trial and error method for the optimization of chromatographic separation conditions but it is not always possible to do so. First the proper HPLC system must be selected and all the parameters of stationary and mobile phase are selected. The compound of interest should take five to ten times longer time to transit through than unretained peak, tM. The type and characteristics of the column packing (porosity, particle size range, good packing procedure, and high quality packing material) also influence column length and the particle size.