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Open AccessArticle

Separation Optimization of a Mixture of Ionized and Non-Ionized Solutes under Isocratic and Gradient Conditions in Reversed-Phase HPLC by Means of Microsoft Excel Spreadsheets

Department of Chemistry, Aristotle University of Thessaloniki, 54124 Thessaloniki, Greece
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Separations 2018, 5(1), 19; https://doi.org/10.3390/separations5010019
Received: 2 February 2018 / Revised: 9 March 2018 / Accepted: 12 March 2018 / Published: 18 March 2018
(This article belongs to the Special Issue Research as Development Perspective)
Τhe crucial role of mobile phase pH for optimizing the separation of a mixture of ionized and non-ionized compounds on a Phenomenex extended pH-range reversed-phase column (Kinetex 5 µm EVO C18) was examined. A previously developed Excel-spreadsheet-based software was used for the whole separation optimization procedure of the sample of interest under isocratic conditions as well as under single linear organic modifier-gradients in different eluent pHs. The importance and the advantages of performing a computer-aided separation optimization compared with a trial-and-error optimization method were realized. Additionally, this study showed that the optimized separation conditions for a given stationary phase may be used to achieve successful separations on new columns of the same type and size. In general, the results of this work could give chromatographers a feel of confidence to establish desired separations of a mixture of ionizable and neutral compounds in reversed-phase columns. View Full-Text
Keywords: reversed-phase liquid chromatography; ionizable and non-ionizable analytes; isocratic and gradient elution in different eluent pHs; computer-assisted separation optimization; visualization of predicted chromatograms reversed-phase liquid chromatography; ionizable and non-ionizable analytes; isocratic and gradient elution in different eluent pHs; computer-assisted separation optimization; visualization of predicted chromatograms
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Zisi, C.; Mangipa, A.M.; Boutou, E.; Pappa-Louisi, A. Separation Optimization of a Mixture of Ionized and Non-Ionized Solutes under Isocratic and Gradient Conditions in Reversed-Phase HPLC by Means of Microsoft Excel Spreadsheets. Separations 2018, 5, 19.

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