Optimization of the separation of (6R)- and (6S)- leucovorin and evaluation of the robustness of the optimum

Abstract The variable-size simplex approach was used to optimize the separation of (6 R )- and (6 S )-leucovorin diastereoisomers. The experimental parameters optimized were pH, ionic strength and percentage of propanol in the mobile phase. The optimization criterion was the valley-to-peak ratio. Fourteen experiments were performed to obtain the optimum. The ruggedness of the optimum was evaluated by means of a partial factorial design for three factors.