Computerized Interpretation of H P L C Chromatogramms by Means of Absorbance Ratio Method and Derivative Spectroscopy
As in other chromatographic processes it is the aim of high pressure liquid chromatography (HPLC) to identify the components of a mixture, whereby there may be some information on the class of substances in question. The analytical procedure can shortly be desribed as follows. The mixture is injected in a mobile phase (solvent) and passes a column where the separation takes place. The separated components leave the column at different retention times. A following detection device generates a signal as a function of concentration (chromatographic peak). Usually the components are identified by relating retention times of external standards to the times of occurence of chromatographic peaks. A System for the computer aided evaluation of such chromatograms was developed by the authors (1). That system was designed in such a way that all components of the mixture to be analyzed are supposed to separate completely, e.g. only chromatograms with distinct peaks were allowed, while overlapping peaks are rejected by a checking algorithm. An inevitable shortcome of this algorithm is, that total overlapping of peaks — which result in a single peak (with undetectable shoulders) — cannot be recognized, so that it was the responsibility of the HPLC- operator to ensure the absence of such envents.
KeywordsHigh Pressure Liquid Chromatography Chromatographic Peak Pure Substance Relate Retention Time Absorbance Ratio
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