Abstract
A rapid chromatographic method for the determination of six phenolic acids (chlorogenic, ferulic, gallic, p-coumaric, protocatechuic and vanillic acids) by ultra performance liquid chromatography (UPLC), was developed and applied for Brazilian tropical fruits mangaba (Hancornia speciosa Gomes) and umbu (Spondias tuberosa Arruda Camara). A multivariate statistical experimental design was employed to optimize analytical conditions (solvent A, solvent B concentrations and flow rates). Samples were cleaned-up by solid-phase extraction (SPE) with different solvents (methanol and acetone) employing SPE cartridges (amine and octadecyl-silane). The method using dihydrogen potassium phosphate 5 mmoL as solvent A and 8% acetonitrile as solvent B presented limits of detection varying from 14 to 94 ng.mL, limits of quantification from 39 to 277 ng.mL-1 with 2 μL of injection volume while total run time for all six compounds was only 9.6 minutes. Higher recovery was obtained by extraction with methanol-acetone of 69.51% to 72.59% for protocatechuic acid and 69.58% to 126.31% for the chlorogenic acid. The concentrations of chlorogenic, p-coumaric and ferulic acids in mangaba extracts were 113.4, 32.1 and 1.5 μg.g-1, respectively while concentrations of chlorogenic, protocatechuic, gallic, vanillic, ferulic and p-coumaric acids present in umbu fruit were 118.9, 141.3, 3.5, 2.5, 2.2 and 1.8 μg.g-1, respectively.
Highlights
Evidences suggest that a diet rich in fruits and vegetables may decrease the risk of chronic diseases, due to the presence of high concentrations of antioxidant compounds, such as phenolic acids [1]
In this paper our objective was to develop an isocratic method for rapid determination of phenolic acids such as chlorogenic, ferulic, gallic, p-coumaric, protocatechuic and vanillic acids using RP-ultra performance liquid chromatography (UPLC) system and to analyze their concentrations in Brazilian tropical fruits of mangaba and umbu
The variables such as flow rate (X1) and the usage of solvent B (X2) were selected based on preliminary studies, which indicated that flow rates higher than 0.6 mL·min−1 implied in lower resolution between all contiguous pairs of peaks (Rs < 1.3)
Summary
Evidences suggest that a diet rich in fruits and vegetables may decrease the risk of chronic diseases, due to the presence of high concentrations of antioxidant compounds, such as phenolic acids [1]. These compounds are widely spread throughout the plant kingdom, occurring in the forms of esters, ethers or free form, and are wellknown for their bioactive properties such as antimutagenic, anticarcinogenic, anti-inflammatory and antimicrobial. The last, carried out in reversedphase mode in conventional high pressure liquid chromatographic system (RP-HPLC) is the most common technique employed and in which the octadecyl-silane (C18) is used as the stationary phase, and diode array as the detector [8].
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