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  • Title: "Orthogonal" separations for reversed-phase liquid chromatography.
    Author: Pellett J, Lukulay P, Mao Y, Bowen W, Reed R, Ma M, Munger RC, Dolan JW, Wrisley L, Medwid K, Toltl NP, Chan CC, Skibic M, Biswas K, Wells KA, Snyder LR.
    Journal: J Chromatogr A; 2006 Jan 06; 1101(1-2):122-35. PubMed ID: 16236292.
    Abstract:
    A general procedure is proposed for the rapid development of a reversed-phase liquid chromatographic (RP-LC) separation that is "orthogonal" to a pre-existing ("primary") method for the RP-LC separation of a given sample. The procedure involves a change of the mobile-phase organic solvent (B-solvent), the replacement of the primary column by one of very different selectivity, and (only if necessary) a change in mobile phase pH or the use of a third column. Following the selection of the "orthogonal" B-solvent, column and mobile phase pH, further optimization of peak spacing and resolution can be achieved by varying separation temperature and either isocratic %B or gradient time. The relative "orthogonality" of the primary and "orthogonal" RP-LC methods is then evaluated from plots of retention for one method versus the other. The present procedure was used to develop "orthogonal" methods for nine routine RP-LC methods from six pharmaceutical analysis laboratories. The relative success of this approach can be judged from the results reported here.
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