Journal
ELECTROPHORESIS
Volume 39, Issue 15, Pages 1993-2000Publisher
WILEY
DOI: 10.1002/elps.201700469
Keywords
Flavanones; LCxLC-PDA-MS; Licorice; Metabolic coverage; Orthogonality; Plant extracts
Funding
- University of Messina through Research and Mobility collaborative project
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Metabolic profiling of Glycyrrhiza glabra using comprehensive two-dimensional liquid chromatography (LCxLC) coupled with photodiode array (PDA) and mass spectrometry (MS) detection is described. The separation was conducted under reversed-phase conditions, using a combination of first dimension (D-1) 150mm microbore cyano column utilising 2.7 m diameter (d(p)) particles, and second dimension (D-2) 50mm superficially porous octadecylsilica column with 2.7m d(p) particles. A multi-segmented shift gradient (MSG) for the D-2 separation was developed, and the orthogonality achieved was compared with other modes of gradients, such as full in-fraction, and shift gradient systems. Results demonstrated a significant expansion of metabolic coverage using MSG in D-2, providing the highest measure of orthogonality compared to other gradient modes. Compound identifications were performed by employing complementary data from PDA and MS detection, with reference to structural group-type distribution in 2D space. A total of ca. 120 compounds were detected, and among them 37 were tentatively identified, distributed over the chemical families of glycosylated flavanones, triterpene saponins, and others. In comparison with one-dimensional LC, the total number of compounds detected was ca. 2-fold greater when LCxLC was employed. To the best of our knowledge, this is the first demonstration of the MSG mode in LCxLC, representing a powerful strategy to expand the metabolic coverage for analysis of plant-derived extracts, containing a multitude of different phytochemical classes.
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