Development and application of a quantitative method based on LC-QqQ MS/MS for determination of steviol glycosides in Stevia leaves.

Stevia is a currently well-known plant thanks to the presence of steviol glycosides, which are considered as sweeteners obtained from a natural source. In this research, a method based on LC-MS/MS by using a triple quadrupole detector was developed for quantitation of 8 steviol glycosides in extracts from Stevia leaves. The ionization and fragmentation parameters for selected reaction monitoring were optimized. Detection and quantitation limits ranging from 0.1 to 0.5ng/mL and from 0.5 to 1ng/mL, respectively, were achieved: the lowest attained so far. The steviol glycosides were quantified in extracts from leaves of seven varieties of Stevia cultivated in laboratory, greenhouse and field. Plants cultivated in field presented higher concentration of steviol glycosides than those cultivated in greenhouse. Thus, the way of cultivation clearly influences the concentration of these compounds. The inclusion of branches together with leaves as raw material was also evaluated, showing that this inclusion modifies, either positively or negatively, the concentration of steviol glycosides.

Tentative identification of polar and mid-polar compounds in extracts from wine lees by liquid chromatography-tandem mass spectrometry in high-resolution mode.

Sustainable agriculture has a pending goal in the revalorization of agrofood residues. Wine lees are an abundant residue in the oenological industry. This residue, so far, has been used to obtain tartaric acid or pigments but not for being qualitatively characterized as a source of polar and mid-polar compounds such as flavonoids, phenols and essential amino acids. Lees extracts from 11 Spanish wineries have been analyzed by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) in high resolution mode. The high-resolution power of LC-MS/MS has led to the tentative identification of the most representative compounds present in wine lees, comprising primary amino acids, anthocyans, flavanols, flavonols, flavones and non-flavonoid phenolic compounds, among others. Attending to the profile and content of polar and mid-polar compounds in wine lees, this study underlines the potential of wine lees as an exploitable source to isolate interesting compounds.

Characterization and Comparison of Wine Lees by Liquid Chromatography-Mass Spectrometry in High-Resolution Mode.

Wine lees from 11 different wineries pertaining to two denominations of origin in Spain (La Rioja and Ribera del Duero) have been characterized in this research by LC-MS/MS in high-resolution mode. For this purpose, the wine lees were separated into the liquid phase (imbibed wine from lees) and the solid residue, which was dried and subjected to solid-liquid extraction assisted by microwaves (dried lees). Both fractions were separately analyzed and the fractions from the 11 wineries compared to find similarity in their patterns. The statistical analysis enabled both differences and common aspects in the composition of imbibed wine from lees and dried lees from all wineries to be found. MS/MS tentative identification of representative compounds in each fraction revealed the varied composition of wine lees with special emphasis on flavonoids such as quercetin, myricetin, and malvidin 3-galactoside, identified in extracts of dried lees, or other compounds such as kaempferol 3-(2',3'-diacetylrhamnoside)-7″-rhamnoside, aminocaproic acid, and citric acid, exclusively identified in imbibed wine from lees. The adsorbent capacity of the solid residue justified the high concentration of phenolic compounds in the extracts from solid lees. The differences found in the composition of the two phases support the separated exploitation of them.

Anthocyanidins, proanthocyanidins, and anthocyanins profiling in wine lees by solid-phase extraction-liquid chromatography coupled to electrospray ionization tandem mass spectrometry with data-dependent methods.

A method has been developed to study the content of anthocyanidins, proanthocyanidicins, and anthocyanins in wine lees, an abundant byproduct from wineries. Detection/quantitation of the target compounds was carried out by a hyphenated system consisting of a solid-phase extraction workstation (Prospekt-2 unit) online coupled to a liquid chromatograph-triple-quadrupole tandem mass spectrometer (LC-MS/MS), where standards were used for identification/quantitation of both anthocyanidins and proanthocyanidins. Owing to the lack of anthocyanins standards, advantages from the use of data-dependent methods were taken for their identification and confirmatory analysis. Combination of the scanning methods (viz. product-ion, precursor-ion, and neutral-loss scanning) allowed identifying five different anthocyanins present in wine residues. The results thus obtained have been validated by complementary analysis of the extracts using LC-TOF/MS in high-resolution mode. Quantitation of the monitored compounds was supported on selected reaction monitoring (SRM) and calibration curves run with standards of anthocyanidins and proanthocyanidins.