A complete package of functions in the R-language has been written for professional separation optimization of complex mixtures of ionized and/or non-ionized solutes. The package includes functions for (a) base-line correction of experimentally recorded chromatograms, (b) modeling of chromatographic peak shapes and retention data, (c) prediction of the retention time of the test analytes and/or their chromatograms, and (d) separation optimization under either isocratic or single and/or double gradient elution conditions by changing the organic modifier(s) content and/or eluent pH. The optimization functions presented in this study offer two different modes for selection of optimal separation conditions: automatic and manual mode. In the automatic mode, the optimal separation conditions are determined by maximizing the resolution within separation time preset by the analyst. In the manual mode, the optimal separation conditions are selected via scatter or contour plots. The foreknowledge of the precise dependence of resolution and separation time upon one or two retention parameters of interest, provided by the proposed computer-assisted separation optimization method, gives chromatographers a feel of confidence for the selection of the optimal conditions for a desired separation. An illustrative video given in the Supplementary material may encourage a novice practitioner in R (software) programming language to follow the proposed separation optimization procedure in a real HPLC analysis.
Keywords: Computer assisted separation optimization and simulation; HPLC analysis; Isocratic and gradient elution; R programming language.
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