Different Extraction Approaches for the Analysis of Melatonin from Cabernet Sauvignon and Feteasca Neagra Wines Using a Validated HPLC-FL Method

Abstract

In recent years, the wine industry has shown a considerable degree of interest in the occurrence of melatonin in wines. Sample pretreatment may be the most important step in trace analysis. Since wine is a complex matrix and melatonin is present in low amounts (ppb), an adequate extraction technique is required. In this study, the effect of several extraction methods, such as solid phase extraction (SPE), Quick, Easy, Cheap, Effective, Rugged, and Safe extraction (QuEChERS), and dispersive liquid–liquid micro-extraction (DLLME) was studied and the variable parameters that can arise throughout the extraction process were optimized to obtain the best results. A high-performance liquid chromatography with fluorescence detector (HPLC-FL) method was adapted and validated, including measurement uncertainty, for the analysis of melatonin in wines and to assess the efficiency of the extraction yield. After comparing the acquired results, the DLLME method was optimized. Extraction recoveries values ranging from 95 to 104% demonstrated that the approach may be successfully applied for the extraction and concentration (enrichment factor of almost eight) of melatonin in wine samples prior to HPLC-FL analysis. The first report of melatonin levels in Feteasca Neagra wines has been made. The data obtained for Cabernet Sauvignon revealed that the final levels of melatonin in the wines are dependent on the winemaking process.

Winemaking Technologies for the Production of Cabernet Sauvignon and Feteasca Neagra Wines Enriched with Antioxidant Active Principles Due to the Addition of Melatonin

“…After vortexing the samples for 10 min at 1200 rpm, 1 g of NaCl salt was added and then mixed again for 5 min. The next step was centrifugation at 5000 rpm for 5 min, after which the analyte of interest was found in the lower phase [ 35 ]. All wine samples were filtered into HPLC vials with a 0.2 µm Syringe Filter Unit (PTFE, Agilent, Beijing, China) and injected into the HPLC system.…”

“…The identification and quantification of melatonin by HPLC-FL were performed using a previously developed method [ 35 ], a Kromasil 100-5-C18 2.1 × 100 mm column, an elution gradient of mobile phase (methanol, solvent A, and 1% acetic acid, solvent B) and flow rate of 1 mL min −1 . The analysis time was 30 min and the column temperature was 20 °C.…”