A fast method using a new hydrophilic–lipophilic balanced sorbent in combination with ultra-high performance liquid chromatography for quantification of significant bioactive metabolites in wines

Abstract

a b s t r a c tThis manuscript describes the development and validation of an ultra-fast, efficient, and high throughput analytical method based on ultra-high performance liquid chromatography (UHPLC) equipped with a photodiode array (PDA) detection system, for the simultaneous analysis of fifteen bioactive metabolites: gallic acid, protocatechuic acid, (−)-catechin, gentisic acid, (−)-epicatechin, syringic acid, p-coumaric acid, ferulic acid, m-coumaric acid, rutin, trans-resveratrol, myricetin, quercetin, cinnamic acid and kaempferol, in wines. A 50-mm column packed with 1.7-m particles operating at elevated pressure (UHPLC strategy) was selected to attain ultra-fast analysis and highly efficient separations. In order to reduce the complexity of wine extract and improve the recovery efficiency, a reverse-phase solid-phase extraction (SPE) procedure using as sorbent a new macroporous copolymer made from a balanced ratio of two monomers, the lipophilic divinylbenzene and the hydrophilic N-vinylpyrrolidone (Oasis TM HLB), was performed prior to UHPLC-PDA analysis. The calibration curves of bioactive metabolites showed good linearity within the established range. Limits of detection (LOD) and quantification (LOQ) ranged from 0.006 g mL −1 to 0.58 g mL −1 , and from 0.019 g mL −1 to 1.94 g mL −1 , for gallic and gentisic acids, respectively. The average recoveries ± SD for the three levels of concentration tested (n = 9) in red and white wines were, respectively, 89 ± 3% and 90 ± 2%. The repeatability expressed as relative standard deviation (RSD) was below 10% for all the metabolites assayed. The validated method was then applied to red and white wines from different geographical origins (Azores, Canary and Madeira Islands). The most abundant component in the analysed red wines was (−)-epicatechin followed by (−)-catechin and rutin, whereas in white wines syringic and p-coumaric acids were found the major phenolic metabolites.The method was completely validated, providing a sensitive analysis for bioactive phenolic metabolites detection and showing satisfactory data for all the parameters tested. Moreover, was revealed as an ultra-fast approach allowing the separation of the fifteen bioactive metabolites investigated with high resolution power within 5 min.

A new and improved strategy combining a dispersive-solid phase extraction-based multiclass method with ultra high pressure liquid chromatography for analysis of low molecular weight polyphenols in vegetables

“…In comparison with other methods to quantify the LMW-PPs, the QuEChERS method offers better selectivity than the SPE Oasis HLB technique [22] and better sensitivity to some analytes than obtained by MEPS C8 [24]. However, regarding the figures of merit of the different techniques, similar results were obtained for LODs, LOQs, trueness and precision.…”

“…This physiological benefits have been attributed to their potential role on the prevention of low-and very-low density lipoprotein (LDL and vLDL) oxidation (crucial steps in atherosclerotic lesion formation) and DNA bases (relevant to the induction of methods for their identification and quantification [14][15][16][17][18]. In recent years, the ability of several extraction techniques, such as solid-liquid extraction (SLE) [19], enzyme-assisted extraction [20], heat extraction [21], solid-phase extraction (SPE) [22], and solid-phase microextraction (SPME) [23], for the isolation free low molecular-weight polyphenols (LMW-PPs) from vegetables and other food matrices, has been proposed and evaluated as reliable alternatives to classic liquid-liquid extraction (LLE) technique. More recently, a novel analytical approach, based on miniaturized microextraction by packed sorbent (MEPS), followed by ultrahigh pressure liquid chromatography (UHPLC) separation, has been proposed by Gonç alves et al [24] for quantitative determination of wine biologically active phenolic constituents and trans-resveratrol [25].…”

A sensitive microextraction by packed sorbent-based methodology combined with ultra-high pressure liquid chromatography as a powerful technique for analysis of biologically active flavonols in wines

“…SPE. The MEPS procedure was compared to SPE reversephase using Oasis HLB extraction cartridges, developed by Silva et al [43]. Briefly, 1 cc (30 mg) Oasis HLB extraction cartridges were conditioned with 1 mL of ethyl acetate and methanol and equilibrated with 1 mL of water.…”

“…Their performance was compared with a previously SPE Oais HLB validated method in our laboratory by Silva et al [43]. The validation parameters are shown in Table 1.…”

“…Many analytical methods have been described for determination of flavonols in wines. The most selective methods involve often liquid or gas chromatography coupled to detection by mass or tandem mass spectrometry [35][36][37][38][39][40][41][42][43][44][45][46][47][48].…”