Procedure for the semi-quantitative analysis of chromatographic traces containing overlapping peaks

Abstract

A manual calculation procedure is described for estimating the amplitude of individual peaks in chromatographic traces where peak overlap occurs. The method is based on the assumptions that the peaks are Gaussian in shape and are reproducible, that small changes in elution volumes can be accommodated by a simple correction procedure and that individual protein elution curves can be summed to give the observed experimental curve. The reproducibility of peak height estimations for five consecutive repeat chromatographic separations ranged from 1.5% for major components to 15% for minor peaks.