A Powerful Analytical Strategy Based on QuEChERS-Dispersive Solid-Phase Extraction Combined with Ultrahigh Pressure Liquid Chromatography for Evaluating the Effect of Elicitors on Biosynthesis of trans-Resveratrol in Grapes

Abstract

A powerful methodological approach based on modified quick, easy, cheap effective, rugged, and safe (QuEChERS) extraction technique, followed by cleanup dispersive solid-phase extraction (dSPE) and combined with ultrahigh pressure liquid chromatography (UHPLC), is presented in this paper, for the rapid determination of the healthpromoting phytoalexin, trans-resveratrol, in grapes. This is the first time, to our knowledge, that the combination QuEChERS-dSPE/UHPLC-PDA has been used for trans-resveratrol quantification in grapes. The method has been validated according to European Union Decision 2002/657/EC, in terms of selectivity, linearity, sensitivity, instrument LOD and method LOQ, accuracy, precision, extraction efficiency, and interference assessment. Validation experiments revealed sufficient linearity (R 2 =0.9931) within the established concentration range confirmed by Mandel's fitting test. The sensitivity was good with method detection limit (LOD) of 0.003 μg/mL and quantification limit (LOQ) of 0.010 μg/mL. Optimum QuEChERS-dSPE/UHPLC-PDA conditions led to recoveries of the target analyte in grape samples ranging between 75.1 and 99.7 % and precisions in the 0.9-4 % range (RSD, n=18). The method also afforded satisfactory results in terms of extraction efficiency and interference assessment and provides a valuable and promising tool for determination of trans-resveratrol in grapes with a high resolution within 8 min and a total analysis time of 11 min. The validated methodology was applied to evaluate the effect of the use of elicitors, namely benzothiadiazole and methyl jasmonate, in trans-resveratrol biosynthesis on Vitis vinifera Monastrell grapes. The results obtained revealed an increase of trans-resveratrol level of 2-fold for grapes treated with benzothiadiazole and 3.5-fold for grapes treated with methyl jasmonate.

An improved analytical method for determination of trans‐resveratrol and related stilbenes in grape skin by QuEChERS coupled with HPLC‐PDA‐MS

“…In a later study, Ruiz‐García et al . (2016) have shown a resveratrol recovery of 93.7–102.5%, LOD of 0.003 μg mL −1 and LOQ of 0.010 μg mL −1 for a QuEChERS method coupled with UHPLC‐PDA by employing acetonitrile:ethyl acetate (50:50, v/v) as extraction solvent, 1 g of trisodium citrate dihydrate, 0.5 g of disodium hydrogen citrate sesquihydrate, 1 g of NaCl and 4 g of MgSO 4 as extraction powder as well as 300 mg of MgSO 4 plus 300 mg of PSA as purification powder.…”

“…Two QuEChERS methods based on Fontana & Bottini (2014) for the determination of polyphenols in wine and Ruiz‐García et al . (2016) for the determination of trans‐resveratrol in grapes were modified for evaluation of extraction and purification efficiency of trans‐resveratrol and related stilbenes in grape skin. The QuEChERS extraction kit (Yo‐Ho Trade Co., New Taipei City, Taiwan) containing extraction powder (4‐g anhydrous magnesium sulphate plus 1‐g anhydrous sodium acetate), purification powder (900‐mg magnesium sulphate plus 10‐, 25‐, 50‐, 100‐ or 300‐mg primary secondary amine [PSA]) and a ceramic homogeniser was used.…”

“…Resveratrol (3,5,6′‐trihydroxy‐trans‐stilbene), an abundant polyphenol in grape skin, is produced as a phytoalexin naturally in response to biotic and abiotic factors. It is biosynthesised from phenylalanine by the shikimic acid pathway involving enzymes such as phenylalanine ammonia‐lyase and stilbene synthetase (Ruiz‐García et al ., 2016). Ever since its first isolation in 1939 by a Japanese scientist Takaoka, numerous studies have demonstrated its vital biological functions including antioxidation, anti‐inflammation, anti‐cancer, anti‐diabetes, anti‐obesity, anti‐microbial, anti‐ageing, cardioprotection and neuroprotection (Lu et al ., 2018; Fiod Riccio et al ., 2020).…”

“…The versatile QuEChERS method is employed in a wide variety of sample matrices and possesses several advantages such as the environment‐ and user‐friendly green approach, low solvent consumption, less lab space and washing requirement, low‐sorbent usage, efficient dispersive sorbent interaction with matrix components for enhancement of signal‐to‐noise ratio, less sample pretreatment time with low‐cost and high‐sample throughput (Kim et al ., 2019). Although the QuEChERS method was used for a wide variety of chemical compounds, their application to bioactives like trans‐resveratrol remains to be less explored (Fontana & Bottini, 2014; Ruiz‐García et al ., 2016; Lu et al ., 2018). Thus, an effective extraction and purification method combined with a sensitive detection technique is essential to explore the application of value‐added byproducts in grape skin.…”

Electrochemical determination of resveratrol in dietary supplements at a boron-doped diamond electrode in the presence of hexadecyltrimethylammonium bromide using square-wave adsorptive stripping voltammetry

“…Due to the above-mentioned interest in RES, several analytical methods have been reported in the literature for the determination of this compound in different matrices. These particularly include high-performance liquid chromategraphy (HPLC), [9][10] ultrahigh pressure liquid chromatography (UHPLC), 11 liquid chromatography coupled to electrospray ionization-mass spectrometry (LC-ESI--MS), 12 gas chromatography-mass spectrometry (GC-MS) 13 and capillary electrophoresis (CE). 14 Electrochemical techniques, such as the voltammetric ones, are the area promising alternatives to classical approaches due to their simplicity, good stability, high sensitivity, speedy procedure and low cost.…”