A simplified description of HPLC separation under overload conditions. A synthesis and extension of two recent approaches

Preparative HPLC resolution of theCIS cyclohexane analogs of phenylalanine

“…UV detection was at 265 nm. Both the column loading capacity, W s (defined as the maximum sample mass that the column can hold), 22 and the optimum sample concentration were calculated for the analytical 150 × 4.6 mm ID column by injecting increasing amounts of sample at different concentrations. In this way, the capacity of the semipreparative column was established as 45 mg and the optimum concentration of the sample as 300 mg/ml.…”

Trends in Preparative HPLC

“…This optimization approach has the advantage of being fast and simple, but it often overlooks specific effects that happen at larger loads. These effects concern the displacement of one product by another and have been described by Guiochon and co-workers [25][26][27][28] and Cox and co-workers [29][30][31]. The interplay of adsorption isotherm, peak form, and capacity factor k during overloading of a column is depicted in Figure 21-3 [32].…”

First synthesis of the two enantiomers of α‐methyldiphenylalanine [(αMe)Dip] by HPLC resolution

“…UV detection was performed at 260 nm (monitoring at lower wavelengths led to saturation of the detector). Both the column loading capacity (Ws, defined as the maximum sample mass that the column can hold) 28 and the optimum sample concentration had been previously determined on the analytical 150´4.6 mm ID column by injecting increasing amounts of sample at different concentrations.…”