Journal
JOURNAL OF SEPARATION SCIENCE
Volume 33, Issue 17-18, Pages 2841-2853Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.1002/jssc.201000274
Keywords
Anthocyanins; Micro-SPE; Plasma; Procyanidins; Ultra-HPLC
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Funding
- Spanish Minister of Industry through Shirota Functional Foods, S.L.
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In the analysis of biological samples, such as plasma or serum, the quantity of sample available is a critical parameter in most cases. A good approach is the use of the microelution SPE (mu SPE) plates as sample pre-treatment technique in which the loaded sample volume is low. An off-line mu SPE and ultra-performance LC-ESI-MS/MS (UPLC-ESI-MS/MS) method was developed and validated to determine procyanidins and anthocyanins in spiked plasma samples. The sample pre-treatment mu SPE allowed the simultaneous determination of procyanidins and anthocyanins from plasma by using a small sample volume (350 mu L) and without an evaporation step previous to the chromatographic analysis. Moreover, the use of UPLC technique allowed to determine the studied compounds at low concentration levels in a short analysis time (12.5 min approximately). Then, the developed method was applied to determine the studied compounds, procyanidins and anthocyanins, and their metabolites in rat plasma samples. Previously, the rats had consumed 5000 mg/kg of a grape pomace extract and the plasma was extracted 4 h after administration. The procyanidins catechin and epicatechin glucuronide, methyl catechin and epicatechin glucuronide, and methyl catechin and epicatechin sulphate were detected at mu M concentration level, and the parent anthocyanins at nM.
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