Exploration of the anthocyanin and proanthocyanidin profile of Greek red grape skins belonging to Vradiano, Limnio, and Kotsifali cultivars, analyzed by a novel LC-QTOF-MS/MS method.

INTRODUCTION: Winegrape varieties Kotsifali, Limnio, and Vradiano OBJECTIVE: The aim of this study was to develop a liquid chromatographic quadrupole time-of-flight tandem mass spectrometric (LC-QTOF-MS/MS) method for the investigation of the anthocyanin and proanthocyanidin content of Greek grape varieties employing target and suspect screening strategies. METHODOLOGY: A novel LC-QTOF-MS/MS method was developed and validated to assess the anthocyanin content of Kotsifali, Limnio, and Vradiano grape varieties. Sixteen grape samples were collected from the main growing areas of each variety in Greece. The influence of the grape variety on the anthocyanin and proanthocyanidin composition of three Greek winegrapes was investigated using chemometrics. RESULTS: Excellent linearity (R2 > 0.99) was achieved for all the target analytes, and recoveries ranged between 90.1% and 119.1%. The limits of quantification (LOQs) and limits of detection (LODs) were calculated over the range of 0.020-0.40 mg/g and 0.010-0.13 mg/g, respectively. The RSD% was lower than 9.1% and 7.3% for intra-day and inter-day studies, respectively, indicating satisfactory trueness and precision. Target and suspect screening resulted in the identification of 5 and 26 anthocyanins, respectively. CONCLUSIONS: Kotsifali variety exhibited a higher concentration of anthocyanins compared with Vradiano and Limnio. Higher levels of mean degree of polymerization (mDp) and different percentage levels of prodelphinidins (%P) were established among the varieties.

An Optimized HPLC-DAD Methodology for the Determination of Anthocyanins in Grape Skins of Red Greek Winegrape Cultivars (Vitis vinifera L.).

A rapid and simple HPLC-DAD analytical method was developed and optimized for the determination of anthocynanins in three red Greek winegrape varieties (Kotsifali, Limnio, and Vradiano). The critical parameters, such as the acidifying solvent and the extraction temperature, which affect the extraction of anthocyanins from the grapes, were studied to find the optimum values. The developed methodology was validated in terms of selectivity, linearity, accuracy, and precision and presented satisfactory results. The limits of quantification (LOQs) ranged between 0.20 mg/kg to 0.60 mg/kg, and the limits of detection (LODs) ranged between 0.06 mg/kg and 0.12 mg/kg. The RSD% of the within-day and between-day assays were lower than 6.2% and 8.5%, respectively, showing adequate precision. The accuracy ranged between 91.6 and 119% for within-day assay and between 89.9 and 123% for between-day assay. Sixteen samples from the main regions of each variety as well as from the official ampelographic collections of Greece were collected during the 2020 growing season and were further analyzed by HPLC-DAD. Notable differences in the anthocyanin content were detected among the cultivars using hierarchical cluster analysis (HCA).