MGuidelines Recommended for Preparative Separation
Development of a preparative separation method usually starts from an analytical separation with an appropriate stationary phase. C18 is the most widely used packing media for preparative separation. The capacity factor, k, of a sample component is recommended in the range of 2.5 and 20. The preparative separation usually overloads the column to achieve highest yield with some sacrifices to resolution. The selectivity and resolution could be optimized by tuning the slope of the operating mobile phase gradient and the mobile phase composition. To maximize the yield and achieve highest purity, the following guidelines are commended:
Sample solubility in the starting mobile phase is critical (>0.1M or >50mg/mL).
Selectivity (?): ?<1.25 should be avoided; allows for minimal overloading (<1 mg/g packing sorbent).
Selectivity (?): ?>1.5 allows overloading up to 15 mg/g packing sorbent).