Desenvolvimento de uma metodologia analítica para determinação de ácidos fenólicos em amostras de azeite de dendê (Elaeis guineensis) por HPLC com detecção simultânea no UV e eletroquímica (homemade)

Abstract

Phenolic substances have become the subject of investigation due to the numerous benefits to human health and the influence in the food quality. In this work, the phenolic acids in homemade and industrialized palm oil were determined. For these analyzes, it was used a high performance liquid chromatograph (HPLC Agilent 1100 series) with a manual 20μL Rheodyne valve, a reverse phase column C18 Inertsil ODS-3, a guard column Inertsil ODS and a UV-VIS detector for multiple wavelengths (UV-VIS MWD). At the end of HPLC waste channel was adapted an electrochemical cell with a capacity of 1.00 mL, with a standard three-electrode configuration, consisting of glassy carbon working electrode, a Ag/AgCl (KCl, sat) reference electrode and stainless steel needle as counter electrode. The electrochemical measurements were made using a μAutolab type III potentiostat, with its data acquisition software (GPES 4.9 version). The detections were performed sequentially, in the same injection. The optimal mobile phase was able to completely separate the analytes by isocratic elution, being composed of an aqueous solution of H3PO4 pH 2.40, containing 7mmol.L-1 of NaClO4 /isopropanol (87:13 v/v). The wavelength used for the UV detection was 225 nm. The amperometric measurements were performed at +1.1 V for all analytes. The recovery of the samples was performed using internal standardization, using o-coumaric acid as internal standard (PI) and m-coumaric acid as standard surrogate (PS). The recovery of PS was over 75% on both detectors for all samples. The instrumental limits for UV detection ranged from 0.019 (syringic acid) to 0.029 mg L-1 (ferulic acid) and quantification limits from 0.063 (syringic acid) to 0.095 mg L-1 (sinapic acid). In the same way, for electrochemical detection (ED) the instrumental limits was found in the range of 0.015 (p-coumaric acid) at 0.079 mg L-1 (ferulic acid), and the quantification limits were between 0.050 (p-coumaric acid) and 0.27 mg L-1 (ferulic acid). The concentration of phenolic acids in homemade palm oil samples ranged from 0.12 to 10.3 mg L-1 (UV detection) and from 1.0 to 11.0 mg L-1 (ED). The concentrations obtained in the industrialized palm oil samples varied from 0.21 to 14.7 mg L-1 (UV detection) and from 3.0 to 11.9 mg L-1 (ED). The HPLC-UV-ED system has shown promise in the analysis of these phenolic acids in this and other complex matrices.