Journal
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
Volume 405, Issue 13, Pages 4627-4638Publisher
SPRINGER HEIDELBERG
DOI: 10.1007/s00216-012-6567-5
Keywords
Flavan-3-ols; Grapeseed procyanidins; HILIC; LCxLC; Mass spectrometry; Procyanidins; Two-dimensional comprehensive LC
Funding
- Ministerio de Educacion y Ciencia [AGL2011-29857-C03-01, CONSOLIDER INGENIO 2010 CSD2007-00063 FUN-C-FOOD]
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In this work, the development and optimization of a new methodology to analyze grape seed procyanidins based on the application of two-dimensional comprehensive LC is presented. This two-dimensional method involves the use of a microbore column containing a diol stationary phase in the first dimension coupled to either a C-18 partially porous short column or a C-18 monolithic column in the second dimension. The orthogonal hydrophilic interaction x reversed phase liquid chromatography (HILICxRP-LC) system is interfaced through a ten-port two-position switching valve. The optimized HILICxRP-LC separation followed by diode array and tandem mass spectrometry detection (HILICxRP-LC-DAD-MS/MS) made possible the direct analysis of a complex grape seed extract and allowed the tentative identification of 43 flavan-3-ols, including monomers and procyanidin oligomers till a polymerization degree of 7 units with different galloylation degrees. To the best of our knowledge, this is the first time that this powerful analytical technique is employed to characterize complex procyanidin samples. This work successfully demonstrates the great capabilities of the HILICxRP-LC-DAD-MS/MS coupling for the direct analysis of very complex natural samples like grape seeds.
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