Journal
JOURNAL OF CHROMATOGRAPHY A
Volume 1181, Issue 1-2, Pages 51-59Publisher
ELSEVIER
DOI: 10.1016/j.chroma.2007.12.034
Keywords
HILIC; comprehensive two-dimensional liquid chromatography; orthogonality; quillaja saponins
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Hydrophilic interaction chromatography (HILIC) was used in two dimensions in a comprehensive two-dimensional HILIC hyphenated with a quadrupole time-of-flight mass spectrometry (HILIC x HILIC-Q-TOF-MS) system for the analysis of complex samples of hydrophilic compounds. A TSKgel Amide-80 column was employed as the first dimension, and a short PolyHydroxyethyl A column was as the second dimension. The column system showed moderate orthogonality at defined operational conditions. A high speed Q-TOF-MS detector as a third complementary dimension significantly improved the peak capacity. The separation capability of the developed HILIC x HILIC-Q-TOF-MS system was tested by separating an extract from Quillaja saponaria. The major components, quillaja saponins, in the extract were well identified by means of [M-H](-) ions, characteristic product ions, and their two-dimensional retention behaviors. Several pairs of isomers, which were often co-eluted on conventional LC-MS methods and had similar fragmentation characteristics in MS/MS spectra, were well separated on the two-dimensional system based on their different hydrophilicity. The developed comprehensive two-dimensional HILIC system demonstrates unique selectivity for hydrophilic compounds and satisfactory peak capacity and resolution for analogues by making sufficient use of two-dimensional separation plane. (c) 2007 Elsevier B.V. All rights reserved.
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