Optimizing Mobile Phase Composition, its Flow Rate and Column Temperature in HPLC Using an Experimental Design Assisted with A Simplex Method

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Abstract
In this work, a rapid procedure to separate ten compounds in high performance liquid chromatography is presented. The use of at experimental design assisted with a simplex method is proposed to separate these compounds with only thirteen chromatographic analyses to select the mobile phase composition, its flow rate and the column temperature. A flow rate of 0.77 ml/min with a percentage of methanol of 49, 95% in the mixture methanol-water and a column temperature of 51.62°C gave the most efficient separation conditions.