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Issue 7, 2010
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Peak picking and the assessment of separation performance in two-dimensional high performance liquid chromatography

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Abstract

An algorithm was developed for 2DHPLC that automated the process of peak recognition, measuring their retention times, and then subsequently plotting the information in a two-dimensional retention plane. Following the recognition of peaks, the software then performed a series of statistical assessments of the separation performance, measuring for example, correlation between dimensions, peak capacity and the percentage of usage of the separation space. Peak recognition was achieved by interpreting the first and second derivatives of each respective one-dimensional chromatogram to determine the 1D retention times of each solute and then compiling these retention times for each respective fraction ‘cut’. Due to the nature of comprehensive 2DHPLC adjacent cut fractions may contain peaks common to more than one cut fraction. The algorithm determined which components were common in adjacent cuts and subsequently calculated the peak maximum profile by interpolating the space between adjacent peaks. This algorithm was applied to the analysis of a two-dimensional separation of an apple flesh extract separated in a first dimension comprising a cyano stationary phase and an aqueous/THF mobile phase as the first dimension and a second dimension comprising C18-Hydro with an aqueous/MeOH mobile phase. A total of 187 peaks were detected.

Graphical abstract: Peak picking and the assessment of separation performance in two-dimensional high performance liquid chromatography

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Publication details

The article was received on 30 Oct 2009, accepted on 09 Apr 2010 and first published on 05 May 2010


Article type: Paper
DOI: 10.1039/B922759H
Analyst, 2010,135, 1541-1550

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    Peak picking and the assessment of separation performance in two-dimensional high performance liquid chromatography

    P. G. Stevenson, M. Mnatsakanyan, G. Guiochon and R. A. Shalliker, Analyst, 2010, 135, 1541
    DOI: 10.1039/B922759H

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