A NEW SEQUENTIAL PROCEDURE FOR THE EFFICIENT AND AUTOMATED LOCATION OF OPTIMUM CONDITIONS IN HIGH PERFORMANCE LIQUID CHROMATOGRAPHY (HPLC)

摘要

A new sequential Optimization Procedure by Search Point (OPSP) based on Hooke-Jeeves algorithm is developed. The procedure is an automated multifactor optimization of conditions for an HPLC separation. Its usefulness in computer-assisted method development is shown by the experimental mobile phase optimization of an isocratic reverse phase liquid chromatography separation of a mixture of six selected pesticides. The relative composition of a ternary mobile phase (Acetonitrile, Methanol, Water) was varied during the optimization process. An objective function (OF) which was used as the criterion of quality of the chromatographic separation is described. The performance of this new chromatographic method is evaluated either by plotting the map of the separation quality using a Grid Search method or by comparing the results with the ones obtained by the application of a Modified Simplex method, in both cases over the same triangular (Acetonitrile, Methanol, Water) parameter space. The optimum solvent composition for the satisfactory separation of pesticides by application of the OPSP was determined from only eleven chromatographic experiments while by applying the Modified Simplex or the Grid Search procedure the number of experiments were 15 and 36 respectively.