Manipulation of mobile phase parameters for the HPLC separation of endogenous monoamines in rat brain tissue.

Multi-component mobile phases are common in ion pair reverse phase high performance liquid chromatography for the analysis of endogenous monoamines in rat brain tissue. By examining the effects of each component of a mobile phase on the separation of a number of monoamines and their metabolites, we show how optimization of parameters can achieve and/or maintain a separation both within and between different octadecyl columns. The assay itself is rapid, sensitive, demands minimal sample preparation, and results in complete resolution of the amines. More importantly, the basis for the manipulation of mobile phase components is discussed with regard to practical utilization in selective amine separation. We conclude that knowledge of mobile phase parameters, their mechanism, and manipulation is as important as the assay itself.