The presented data illustrate how an LC–MS-compatible separation of four hydrophilic test solutes in 12 min can be accomplished
using ANP chromatography. Using a quick six-step process, the final method was devised that produced both good separation
and symmetrical peak shapes. Before each step, predictions and expectations were made on how the analytes would interact under
the method conditions studied based on the appropriate retention model. After each step, the results were scrutinized in terms
of their agreement with the model and the predictions made from it. Further predictions using a better understanding of the
retention mechanism could then be used to refine the method. The differences observed in the data are consistent with what
is known about ANP retention and how the ionization state of both the analytes and the stationary phase can affect it. Keeping
these factors in mind for prediction of chromatographic behavior leads to a much more streamlined method development process.
Table IV: Parameters investigated for robustness
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Joshua E. Young and Maria T. Matyska
are with MicroSolv Technology Corporation in South New Berlin, New York.
Hong Nhung Nguyen and Joseph J. Pesek
are with the Department of Chemistry at San José State University in San José, California. Direct correspondence to: firstname.lastname@example.org