QUANTITATIVE RESOLUTION OF SEVERELY OVERLAPPING HIGH PERFORMANCE LIQUID-CHROMATOGRAPHIC PEAKS

Severely overlapping high performance liquid chromatographic peaks have been resolved with a mathematical technique to extract quantitative information from unresolved peaks. Peak shapes must differ if the retention volumes are identical, or the retention volumes must be slightly different if the peak shapes are identical. Chromatographic peak positions must be corrected for changes in the mobile phase flow rate. A mobile phase flow rate monitor and the associated computer routines were developed to make the necessary corrections. Overlapping synthetically generated peaks and experimentally measured chromatograms were resolved to demonstrate the applicability of this technique and the necessity for flow corrections. Minor changes in the retentive ability of the chromatographic column required a correction procedure to align the peaks for a calibration procedure. Peaks exhibiting a shoulder were resolved with less than 13% error and quantitative information was obtained from completely overlapped peaks with less than 18% error.