New Advice on an Old Topic: Buffers in Reversed-Phase HPLC
Buffers are commonly used in reversed-phase liquid chromatography (LC) to control the ionization state of analytes. However, the addition of buffers is much more complex than simple pH control. Complex equilibria exist between these mobile-phase additives, the analytes, the silica surface, and even the stationary phase in certain circumstances. The addition of mass spectrometry (MS) as a primary detection technique makes decisions about mobile-phase additives even more crucial. In this column instalment, we use a model set of analytes and selected applications to demonstrate the effects that buffers can have not only on the selectivity of a separation, but also on the sensitivity of a reversed-phase analysis when using MS detection.