Phytochemical Screening of Ultrasonic Extracts of Salix Species and Molecular Docking Study of Salix-Derived Bioactive Compounds Targeting Pro-Inflammatory Cytokines.

Willow bark (Salix spp., Salicaceae) is a traditional analgesic and antirheumatic herbal medicine. The aim of this study was to evaluate and compare the phytochemical and antioxidant profiles of leaf and bark extracts of six species of the genus Salix obtained by ultrasound-assisted extraction (UAE) and to examine the inhibitory potential of target bioactive compounds against two inflammatory mediators, tumor necrosis factor alpha (TNF-α) and interleukin 6 (IL-6), through in silico molecular docking. The total phenolic and flavonoid content of the extracts was estimated using spectrophotometric methods and the antioxidant activity using 2,2-diphenyl-1-picrylhydrazyl (DPPH•) and hydroxyl radical (•OH) scavenging assays. Chemical profiling of extracts was carried out using high-performance liquid chromatography coupled with a diode array detector (HPLC-DAD). Principal component analysis (PCA) was performed to differentiate the sample extracts based on their phytochemical profiles and amounts of target bioactive compounds. Chemical composition varied among the analyzed willow species and also among the plant organs of the same species. The major bioactive compounds of the extracts were salicin, chlorogenic acid, rutin and epicatechin. The extracts exhibited significant DPPH● and ●OH scavenging activities. Results of molecular docking revealed that chlorogenic acid had the highest binding affinity toward TNF-α and IL-6. UAE extracts represent valuable sources of antioxidant and anti-inflammatory compounds.

Ibuprofen as an Organic Pollutant in the Danube and Effects on Aquatic Organisms.

The presence of emerging substances in surface water is of a great concern knowing they are the main source for community water supply needs. This study describes the development, optimization and application of an analytical method for the determination of ibuprofen in the Danube samples. Caffeine concentrations, as an indicator of human waste, were determined and maximum risk indexes for aquatic organisms were calculated. The Danube samples were collected from ten representative locations. A Solid-phase extraction was used for ibuprofen and caffeine separation and the analysis was performed by High-performance liquid chromatography method. Ibuprofen concentrations ranged (30.62-111.40) ng/L and caffeine (305.94-375.97) ng/L. Low risk on aquatic organisms was determined for ibuprofen and potential sublethal effect for caffeine was obtained. The results indicated that ibuprofen was effectively separated from other substances in the samples under defined chromatographic conditions for short period of time (4 minutes). Applied HPLC method showed good repeatability, accuracy, selectivity and robustness. Further studies including continuous monitoring of caffeine in the Danube are necessary in order to assess the real risks and possible prevention.

Synthesis and evolution of physicochemical properties of new pharmaceutically active ionic liquids - tetracainium salicylate and tetracainium ibuprofenate.

Tetracainium salicylate and tetracainium ibuprofenate were synthesized as active pharmaceutical ingredient ionic liquids (API-ILs). These ILs represent a combination of a drug for local anaesthesia (tetracaine) and nonsteroidal anti-inflammatory drugs (salicylic acid and ibuprofen). After IL synthesis, spectroscopic investigations were performed using infrared and nuclear magnetic resonance spectroscopy to confirm their structures. Differential scanning calorimetry and thermogravimetric analysis determined the obtained thermal behaviour of the ionic liquids. Experimental density, viscosity, and electrical conductivity measurements were performed in a wide temperature range to understand the interactions occurring in the obtained pharmaceutically active ionic liquids. All experimental values were well-fitted by the empirical equations. According to the theoretical calculations, weaker interactions of tetracaine with ibuprofenate than with salicylate are found, ascribed to the decreasing molecular symmetry, weakened hydrogen bonding, and increasing steric hindrance of ibuprofenate's alkyl chain.

Pharmacological effects of novel microvesicles of basil, on blood glucose and the lipid profile: a preclinical study.

Microencapsulation represents a process that can create targeted, controlled release kinetics of drugs, thus optimizing therapeutic efficacy. Our group has investigated the impact of this technology on Wistar rats to determine pharmacological efficacy of basil extracts. Animals were treated with water extract of Ocimum basilicum in microvesicles and with combination of basil extracts and 3α,7α-dihydroxy-12-keto-5-cholanate, also known as 12-monoketocholic acid (MKC) acid in microvesicles for 7 days. Alloxan was used to induce hyperglycemia. Pharmacological effects on glycemia were evaluated by measuring blood glucose levels in alloxan-induced diabetic rats. Microvesicles were prepared using the Büchi-based microencapsulating system developed in our lab. The dose of basil extract that was orally administered in rats was 200 mg/kg and the dose of MKC acid was 4 mg/kg as per established protocols. A seven-day treatment with basil aqueous extract, as well as a combination of basil and MKC acid extract in the pharmaceutical formulation, led to a statistically significant reduction in the blood glucose concentration of animals with alloxan-induced hyperglycemia compared to pre-treatment values (p < 0.05 and p < 0.01), which indicates that basil has hypoglycemic and antihyperglycemic effects. Microvesicles, as a pharmaceutical-technological formulation, substantially enhance the hypolipidemic action of basil extract with MKC acid.

Experimental and Computational Evaluation of Extraction Procedure and Scavenging Capacity of Sweet Basil Extracts (Ocimum basilicum L.).

The possibility to prevent nutrition-related diseases that include scavenge of free radicals and to block chain reactions is very important and significant for human well-being. The aim of this study was to analyse different basil extracts, determine the relationship between total phenolic/flavonoid content and antioxidant activity in order to optimize its application in industry. The extraction involved different solvents (ethanol, methanol and water), extraction time (10 and 30 min and 24, 48 and 72 h), plant fragmentation level (0.3 and 2 mm) and the presence or absence of light. Antioxidant activity was investigated by applying spectrophotometric method and measuring the total phenolic and flavonoid content and DPPH radical scavenging activity. The content of total phenolics varied from 5.2 to 185.6 mg of gallic acid equivalents per gram of a dry extract and flavonoids ranged from 0.2 to 35.0 mg of quercetin per gram of a dry extract. All extracts presented a scavenging capacity and IC50 values of DPPH radical inhibition ranged from 0.04 to 12.99 µg/ml. The evaluation of experimental data for eighty basil extracts was performed by chemometric analysis showing good correlation between yield and total phenolic compounds, as well as flavonoid content and inhibition of the DPPH radical.

Methodological Aspects of Extraction, Phytochemical Characterization and Molecular Docking Studies of Salix caprea L. Bark and Leaves.

Contents of twelve selected bioactive substances and antioxidant potential of Salix caprea L. extracts were compared in its two vegetative organs (bark and leaves) and in terms of different ethanol/water mixtures used for extraction (30-70% aq, ethanol) and extraction time (30 min; 24, 48 and 72 h). The extracts were characterized by High Pressure Liquid Chromatography (HPLC), and total phenolics and flavonoids were determined spectrophotometrically. All secondary metabolites identified in Salix caprea L. extracts (gallic, chlorogenic and vanillic acid, epicatechin, rutin, quercetin and naringenin) were found more accumulated in bark. Salicin and p-hydroxybenzoic acid were detected in bark and ferulic, trans-cinnamic and p-coumaric acid in leaves extracts only. Rutin was most abundant bioactive compound both in bark (1.71 g/100 g of de) and leaves extracts (0.434 g/100 g of de). Bark extract with highest bioactive substances contents was obtained with 70% aq. ethanol as most suitable solvent during extraction time of 48 h. Molecular docking showed salicin to have similar affinity toward COX-2 as acetylsalicylic acid, but lower toward COX-1.

The occurrence of selected xenobiotics in the Danube river via LC-MS/MS.

Having in mind that there is a general lack of monitoring plans and precaution measures in the developing countries and that the Danube is the second longest river in Europe, the estimation of the relevant concentration levels of unregulated xenobiotics is a topic of interest both on local and international level. The selected pharmaceuticals, caffeine, and benzotriazole presented in the collected water samples from seven representative locations around the territory of Novi Sad, Serbia, during 1-year period, were analyzed with the use of solid-phase extraction followed by the liquid chromatography coupled with triple quad tandem mass spectrometry. The most frequently detected compounds were caffeine and carbamazepine in the concentrations up to 621 and 22.2 ng/L, respectively, while the maximum concentration of the analyzed pharmaceuticals was obtained for ibuprofen (60.1 ng/L). The presence of benzotriazole along the analyzed section of the river was confirmed in the concentration levels up to 26.7 ng/L. Although sulfamethoxazole and desmethyldiazepam were detected at trace levels (0.22 and 3.41 ng/L, respectively); the presence of these pharmaceuticals in complex mixtures should not be neglected. Due to the frequent detection caffeine, carbamazepine, ibuprofen, and benzotriazole could be proper candidate for hydrophilic anthropogenic markers for quantification of wastewater contamination in surface water in the analyzed Danube section.