Determination of Peak Purity in HPLC by Coupling Coulometric Array Detection and Two-Dimensional Correlation Analysis.

This work aims to evaluate the purity of chromatographic peaks by a two-dimensional correlation (2D-corr) analysis. Such an analysis leads to two contour plots: synchronous and asynchronous. The synchronous contour plot provides information on the number of peaks present in the chromatogram. The asynchronous contour plot reveals the presence of overlapping species on each peak. The utility of 2D-corr analysis was demonstrated by the chromatographic analysis of Capsicum chili extracts obtained by HPLC coupled with a coulometric array of sixteen detectors. Thanks to 16 electrochemical sensors, each poised at increasing potentials, the resulting 2D-corr analysis revealed the presence of at least three species on the peak located at a retention time of 0.93 min. Mass spectrometry (MS) analysis was used to analyze the coeluting species, which were identified as: quinic acid (3.593 min), ascorbic acid (3.943 min), and phenylalanine (4.229 min). Overall, this work supports the use of 2D-corr analysis to reveal the presence of overlapping compounds and, thus, verify the signal purity of chromatographic peaks.