Changing reversed-phase high performance liquid chromatography selectivity. Which variables should be tried first?

J Chromatogr B Biomed Sci Appl. 1997 Feb 7;689(1):105-15. doi: 10.1016/s0378-4347(96)00351-9.

Abstract

When carrying out HPLC method development, it is often necessary to vary the relative retention of the sample (values of alpha) by changing some experimental variable, e.g., solvent type, pH, etc. The choice of which variable will be most suitable for a change in selectivity depends on two conflicting goals: (a) the attainment of maximum changes in alpha for the better control of resolution and (b) the avoidance of practical problems associated with the use of a given variable to optimize selectivity. This study provides a quantitative evaluation of different variables for their effect on selectivity (alpha). Various practical problems which must be balanced against this ability of a variable to change value of alpha are also discussed. The selection of any two variables for their simultaneous use in controlling alpha is also examined.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Chromatography, High Pressure Liquid / instrumentation
  • Chromatography, High Pressure Liquid / methods*
  • Hydrogen-Ion Concentration
  • Osmolar Concentration
  • Sensitivity and Specificity
  • Solvents
  • Temperature

Substances

  • Solvents