A systematic way to develop HPLC methods consists of building up and running a useful set of experiments (DOE, design of experiments) to evaluate the influence of several factors or method conditions on the selectivity of a chromatogram. The crucial part here is the peak tracking between the chromatograms. It has been shown that under most conditions peak areas can be used to identify peaks of the same compound in different chromatograms. However, this technique also has some limitations that will be discussed in this article.
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