Analysis of basic compounds by reversed-phase liquid chromatography-electrospray mass spectrometry in high-pH mobile
1Phenomenex, 411 Madrid Avenue, Torrance, CA, USA. limingp@phenomenex.com
Abstract:
The separation of basic compounds in high-pH mobile phases results in extended retention, excellent peak shapes and good chromatographic efficiency. A severe decrease in sensitivity with electrospray mass spectrometric detection in positive ion mode (ESI+-MS) is expected under conditions that suppress analyte ionization in solution. We compared the responses of a large number of various basic drugs covering a wide range of hydrophobic (log P 0.09 to 7.6) and basic character (pKa 6.8-10) in LC-ESI+-MS/MS in 0.1% formic acid in water and acetonitrile, to responses in 10mM ammonium hydrogencarbonate buffers of different pH (7.8-11), and acetonitrile. Contrary to common expectations, high-pH mobile phases do not affect negatively the responses of basic compounds in ESI+. Analyte responses and limits of detection are comparable, or most often better in high pH compared to acidic mobile phases.
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