Peak capacity estimation in isocratic elution

Peak capacity (i.e. maximal number of resolved peaks that fit in a chromatographic window) is a theoretical concept with growing interest, but based on a situation rarely met in practice. Real chromatograms tend to have uneven distributions, with overlapped peaks and large gaps. The number of resolved compounds should, therefore, be known from estimations. Several equations have been reported for this purpose based on three perspectives, namely, the intuitive approach (peak capacity as the size of the retention time window measured in peak width units), which assumes peaks with the same width, and the outlines of Giddings and Grushka, which consider changes in peak width with retention time. In this work, the peak capacity concept is discussed and three new approaches are derived based on realistic descriptions of peak shape. The first one is based on the Grushkaʹs approach and considers the contributions of column and extra-column peak variances. The second one relies on Giddings’ and assumes asymmetrical peaks where left and right peak half-widths depend linearly on retention time. The third equation, based on the intuitive approach, uses a mean peak width obtained by integration, instead of a mean value from several representative peaks. The accuracy of the classical Giddings’ approach for ideal peaks, a modification of the Grushkaʹs approach that considers variation of peak width at half-height, and the three new approaches were checked on combined chromatograms built by adding real peaks. The results demonstrate that the change in efficiency (and not in skewness) is the relevant factor, at least in the studied examples. Also, peak width should be measured at low peak height ratios (i.e. 10%) to better account peak deformation.