Development and validation of a HPLC method using a monolithic column for quantification of trans-resveratrol in
Ana Rute Neves1, Salette Reis1, Marcela A Segundo1
1UCIBIO, REQUIMTE, Department of Chemical Sciences, Faculty of Pharmacy, University of Porto, Rua de Jorge Viterbo Ferreira 228, 4050-313 Porto, Portugal.
Abstract:
The development of nanodelivery systems that protect trans-resveratrol is extremely important to preserve its bioactive properties in the development of further applications as nutraceuticals to supplement foods and beverages. In this work, a validated HPLC method was developed for the quantification of trans-resveratrol in lipid nanoparticles for application in studies of in vitro intestinal permeability. The chromatographic separation was achieved in a C18 monolithic column connected to a fluorometric detector (330/374 nm), by isocratic elution consisting of 2% acetic acid/acetonitrile (80:20). Two calibration ranges were established (0.020-0.200 and 0.200-2.00 μmol L(-1)), and low quantification limits (2-6 nmol L(-1), 23-69 pg) were achieved. Stability studies showed that trans-resveratrol is stable for 24 h at 4 °C, and storage at room temperature and freeze-thaw cycles are not recommended. The proposed method was applied to in vitro intestinal permeability studies, in which values between 0.05 ± 0.01 and 1.8 ± 0.3 μmol L(-1) were found.
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