Macular carotenoids in lipid food matrices: DOE-based high energy extraction of egg yolk xanthophylls and quantification through a validated APCI(+) LC-MS/MS method.

Lutein and zeaxanthin exhibit significant biological activities therefore their dietary intake through carotenoid-rich foods and supplements is strongly recommended as preventive approach. Hence their extraction from natural substrates targets to their commercial exploitation as nutraceuticals and ocular pharmaceuticals. Since carotenoids' bioavailability is higher in fat-containing substrates, egg yolk is considered an ideal food matrix. DOE-based optimization of novel high energy extraction practices achieves efficient recovery of xanthophylls from lipid sources. In this research, 23 full factorial and Box-Behnken designs (BBD) were applied for optimizing ultrasound- (UAE) and microwave-assisted extraction (MAE) variables (i.e. extraction solvent, temperature, time, US or MW power and solvent/material ratio). LC-MS/MS results pointed out the precedence of UAE in lutein and zeaxanthin extraction, where higher yields were obtained with 1:1 n-hexane-acetone as solvent mixture at 19 min, 600 W and 35 mL g-1. UAE carotenoid content was higher than MAE due to the different mechanisms laying behind the two processes and due to more complete granule rupture caused by higher US power. Evaluating the current results, DOE-based UAE analytical methodology stands out as an auspicious and sustainable alternative for commercial-based extraction of lipidic bioactive compounds for food and drug industrial applications.

Binding and stabilisation effects of glycodendritic compounds with peanut agglutinin.

A number of new polyhydroxy-dendritic structures have been constructed from a few basic modules. The cores were derived from N-tert(butyloxycarbonyl)tris[(propargyloxy)methyl]aminomethane, N,N'-bis-1,3-(tris-(propargyloxymethyl)methyl)-5-(hydroxymethyl)isophthalamide, and N,N',N″-tris-1,3,5-(tris-(propargyloxymethyl)methyl)-1,3,5-benzene tricarboxamide while the terminal groups were derived from ß-azido-galactose and ß-azido-lactose leading to six new glycodendrimeric compounds with up to 63 hydroxyl groups on the periphery. The binding ability of the new compounds to peanut agglutinin (PNA), a galactose recognizing lectin from Arachis hypogaea, was investigated by nano-Isothermal Titration Calorimetry and nano-Differential Scanning Calorimetry. We found that the compounds had stronger stabilising effect on the macromolecules compared to the corresponding sugars. The interaction between lectin and the glycodendrimeric unit is entropically driven with only a low enthalpic contribution. A trend was found with increasing number of carbohydrates that is strongly influenced by the steric constraints of the ligands. Our results indicate the significance of multivalency and size control in the successful design of lectin inhibitors.

Response surface methodology toward the optimization of high-energy carotenoid extraction from Aristeus antennatus shrimp.

High-energy assisted extraction techniques, like ultrasound assisted extraction (UAE) and microwave assisted extraction (MAE), are widely applied over the last years for the recovery of bioactive compounds such as carotenoids, antioxidants and phenols from foods, animals and herbal natural sources. Especially for the case of xanthophylls, the main carotenoid group of crustaceans, they can be extracted in a rapid and quantitative way with the use of UAE and MAE. Response surface methodology (RSM) is used for the optimization of extraction methodologies, also being applied to optimize high energy techniques. Three independent variables, namely extraction time, ultrasound or microwave power and solvent/material ratio, were investigated for both methods by employing a 16-run three-level Box-Behnken design (BBD). Considering the extraction efficiency for carotenoids from Aristeus antennatus shrimp, the selected conditions for UAE were 5 min, 600 W and 10:1 mL g(-1). Acetone was the solvent of choice for the extraction procedure. For MAE, the best experimental values were 7 min, 30 W and 20:1 mL g(-1) using n-hexane:acetone:ethanol 2:1:1 (v/v/v) as extraction solvent. The determination of total carotenoid yield was carried out using the spectophotometric calibration curve (A=0.1646(±0.0061)C-0.005(±0.022), R(2)=0.996, n=3) of a standard mix solution of canthaxanthin, zeaxanthin and lutein at 452.1 nm. Under the selected conditions, the yield of total carotenoids for UAE was 23.4(±2.3) and 6.73(±0.56) mg of carotenoids per 100 g dry sample for the head and the body of shrimp, while for MAE was 23.92(±0.63) and 13.3(±1.1) mg of carotenoids per 100g dry sample, respectively. The recovery of both methods was calculated between 60 and 105%. The results indicate that high-energy extraction techniques are faster, less laborious, more repeatable and reproducible methods than the conventional approaches for the quantitative recovery of sensitive bioactive compounds. Moreover, the recovery of a high-added value group of bioactive molecules from natural sources, such as carotenoids, can constitute a profitable and valuable commercial alternative, as these compounds can be used as dietary supplements, food color enhancers and additives in animal feeds, functional foods, preservatives, pharmaceuticals and cosmetics.

Microwave-assisted synthesis of 3,5-disubstituted isoxazoles and evaluation of their anti-ageing activity.

One-pot uncatalysed microwave-assisted 1,3-dipolar cycloaddition reactions between in situ generated nitrile oxides and alkynes bearing protected antioxidant substituents, were regioselectively afforded 3,5-disubstituted isoxazoles. The yields were moderate, based on the starting aldehydes, while the reaction times were in general shorter than those reported in the literature. The cytoprotective and anti-ageing effect of the final deprotected compounds was evaluated in vitro, on cellular survival following oxidative challenge and in vivo, on organismal longevity using the nematode Caenorhabditis elegans. The activity of the isoxazole analogues depends on the nature and the number of the antioxidant substituents. Analogue 17 bearing a phenolic group and a 6-OH-chroman group is a promising anti-ageing agent, since it increased survival of human primary fibroblasts following treatment with H2O2 and extended C. elegans lifespan.

FeCl3-catalysed ultrasonic-assisted, solvent-free synthesis of 4-substituted coumarins. A useful complement to the Pechmann reaction.

The catalytic activity of FeCl3 for the synthesis of a variety of 4-substituted coumarins using high energy techniques has been investigated. The ultrasonic-assisted conditions provide a useful complement to the Pechmann reaction, affording the coumarin derivatives in excellent yields, under solvent-free conditions, in short reaction times using an inexpensive, mild and benign Lewis acid catalyst.

Synergistic effect of dual-frequency ultrasound irradiation in the one-pot synthesis of 3,5-disubstituted isoxazoles.

Herein is reported a one-pot three-step process for the regioselective synthesis of 3,5-disubstituted isoxazoles based on copper(I)-catalyzed cycloaddition reaction between in situ generated nitrile oxides (from the corresponding aldehydes) and alkynes, using ultrasound irradiation, avoiding toxic reagents and solvents and isolation/purification of intermediates. The combined use of 40 kHz ultrasonic bath and 20 kHz probe in the presence of copper turnings reduced reaction time to 1h and resulted in only one final purification step with increased yields, clearly indicating that there is a dual-frequency synergistic effect. In addition, under metal free conditions, the 1,3-dipolar cycloaddition was regioselective giving low to modest yields.

Aryllithiums with increasing steric crowding and lipophilicity prepared from chlorides in diethyl ether. The first directly prepared room-temperature-stable dilithioarenes.

A convenient procedure has been developed for the preparation of synthetically useful, room-temperature-stable aryllithiums starting from aryl chlorides and lithium metal. The method provides a route to aryllithiums which have previously not been accessible cleanly or could only be prepared by using more expensive starting materials.