Saša Savić

Enzymatic oxidation of rutin by horseradish peroxidase: Kinetic mechanism and identification of a dimeric product by LC-Orbitrap mass spectrometry

Flavonoid oxidation is important issue in food processing and quality. The kinetic mechanism of enzymatic oxidation of rutin by horseradish peroxidase (HRP) was studied. Rutin oxidation reaction was followed by recording of spectral changes over the time at 360 nm. The studied oxidation is mostly enzymatic and less part non-enzymatic. The reaction with HRP has a higher rate compared with the reaction without of HRP, whereby is part of non-enzymatic reaction about 10% of the total reaction. Kinetic parameters were determined from graphics of linear Michaelis-Menten equation, and it was found that investigated reactions of rutin oxidation by HRP take place in a ping-pong kinetic mechanism. High resolution HPLC-MS analysis of the mixture of oxidized products of rutin revealed the presence of rutin dimer. Because of widely distribution of rutin as well as presence of peroxidases and hydrogen peroxide in fresh foods identification of this enzymatic modification product can be beneficial for foods quality and safety.

Comparative analysis of antioxidant, antimicrobiological and cytotoxic activities of native and fermented chamomile ligulate flower extracts

AbstractMain conclusionThe work investigated differences in apigenin content, as well as in other compounds, and examined the chemical profiles, antioxidant, antimicrobial and cytotoxic effects of extracts obtained from native and fermented chamomile ligulate flowers. Chamomile (Chamomilla recutita L.) has a long history of being used as a medicinal plant due to many health benefits, including antiinflammatory, anticancer, antispasmodic, radical-scavenging effects and others. Apigenin is recognized as one of the most bioactive phenolic compounds in chamomile. In comparison to its bound forms, which include mostly apigenin-7-O-β-glucoside and various acylated forms, the aglycone is attributed with much higher bioactivity. Due to this fact, in this work ligulate florets of chamomile anthodium were subjected to a fermentation process using native chamomile enzymes to hydrolyze bound forms of apigenin to free aglycone. The contents of apigenin and apigenin-7-O-β-glucoside were determined in both fermented and nonfermented samples by UHPLC-MS–MS analysis to define the efficiency of conversion. After defining their chemical profiles, the extracts of fermented and nonfermented chamomile samples were also compared with respect to their antioxidant, antimicrobial and cytotoxic effects. The antioxidant effects of the obtained extracts were defined by electron spin resonance analysis for hydroxyl and superoxide radicals. The antimicrobial activity was defined for eight microbial strains, whereas cytotoxic activity was evaluated using two human cell lines (human cervix carcinoma and human rhabdomyosarcoma) and murine fibroblasts.

Parenteral nanoemulsions as promising carriers for brain delivery of risperidone: Design, characterization and in vivo pharmacokinetic evaluation.

This paper describes design and evaluation of parenteral lecithin-based nanoemulsions intended for brain delivery of risperidone, a poorly water-soluble psychopharmacological drug. The nanoemulsions were prepared through cold/hot high pressure homogenization and characterized regarding droplet size, polydispersity, surface charge, morphology, drug-vehicle interactions, and physical stability. To estimate the simultaneous influence of nanoemulsion formulation and preparation parameters--co-emulsifier type, aqueous phase type, homogenization temperature--on the critical quality attributes of developed nanoemulsions, a general factorial experimental design was applied. From the established design space and stability data, promising risperidone-loaded nanoemulsions (mean size about 160 nm, size distribution <0.15, zeta potential around -50 mV), containing sodium oleate in the aqueous phase and polysorbate 80, poloxamer 188 or Solutol(®) HS15 as co-emulsifier, were produced by hot homogenization and their ability to improve risperidone delivery to the brain was assessed in rats. Pharmacokinetic study demonstrated erratic brain profiles of risperidone following intraperitoneal administration in selected nanoemulsions, most probably due to their different droplet surface properties (different composition of the stabilizing layer). Namely, polysorbate 80-costabilized nanoemulsion showed increased (1.4-7.4-fold higher) risperidone brain availability compared to other nanoemulsions and drug solution, suggesting this nanoemulsion as a promising carrier worth exploring further for brain targeting.

Antioxidant activity of strawberry (Fragaria × ananassa Duch.) leaves

ABSTRACT Antioxidant activity of aqueous extracts from two cultivated strawberry leaves, Senga Sengana and Mount Everest, obtained by maceration, reflux extraction and ultrasonic extraction, was investigated. Antioxidant activity of extracts was assessed by DPPH, FRAP, FIC and H2O2 test; total phenolics and flavonoids were assessed by Folin–Ciocalteu and aluminium chloride methods, respectively. Extract obtained from Senga Sengana by reflux extraction, containing higher amounts of extractive matter, total phenolics and flavonoids, has shown higher antioxidant activity (EC50(DPPH) = 10.4 μg/mL; FRAP = 258.61 mgFe2+/g dry extract.; FIC activity = 62.6% in concentration of 2 mg/mL; H2O2 scavenging = 25.43% in concentration of 0.5 mg/mL) compared to extracts obtained by other extraction techniques. Rutin, coumaroyl-galloyl-glucopyranoside, hydroxyphenylethyl-coumaroyl-glucopyranoside, derivates of hydroxyphenylethyl-coumaroyl-glucopyranoside, chlorogenic acid and p-coumaroylhexose have been identified by UHPLC–DAD–HESI–MS analysis.

Sulfanilamide in solution and liposome vesicles; in vitro release and UV-stability studies

The main goal of this study was to develop a liposome formulation with sulfanilamide and to investigate the liposomes impact on its release and stability to the UV-A/UV-B and UV-C irradiation. Liposome dispersions with incorporated sulfanilamide were prepared by thin-film hydration method and liposomes role to the sulfanilamide release was investigated by using a dialysis method. Comparatively, sulfanilamide in phosphate buffer solution was subject to release study as well to the UV irradiation providing for the possibilities of kinetics analysis. In vitro drug release study demonstrated that 20% of sulfanilamide was released from liposomes within 1 h that is approximately twice as slower as in the case of dissolved sulfanilamide in phosphate buffer solution. The kinetic release process can be described by Korsmeyer–Peppas model and according to the value of diffusion release exponent it can be concluded that drug release mechanism is based on the phenomenon of diffusion. The sulfanilamide degradation in phosphate buffer solution and liposomes is related to the formation of UV-induced degradation products that are identified by UHPLC/MS analysis as: sulfanilic acid, aniline and benzidine. The UV-induced sulfanilamide degradation in the phosphate buffer solution and liposome vesicles fits the first- order kinetic model. The degradation rate constants are dependent on the involved UV photons energy input as well as sulfanilamide microenvironment. Liposome microenvironment provides better irradiation sulfanilamide stability. The obtained results suggest that liposomes might be promising carriers for delayed sulfanilamide delivery and may serve as a basis for further research.

Synthesis and Physicochemical Characterization of Anion Exchanger Based on Green Modified Bottle Gourd Shell

The structural performance and biosorption behavior of a new cationic biosorbent (CALV) prepared from bottle gourd (Lagenaria vulgaris) shell (LVS) as a potentially valuable agrowaste have been established. The biosorbent with anion exchangeable function was synthesized from a modified lignocellulosic biomass (MLV) by a quaternary ammonium agent, N-(3-chloro-2-hydroxypropyl)trimethylammonium chloride. Preparation of the appropriate MLV precursor was firstly carried out by the process of alkaline pretreatment of LVS biomass using a green carbonate solution. In this way, partial delignification, removal of extractive substances, and increased porosity of the cellulose-enriched MLV biomass, as well as the activation of available cellulosic microfibrils to alkaline-cellulose have been achieved. The optimization of synthesis conditions was carried out by direct estimation of the CALV biosorption efficiency to phosphate (using ICP-OES) and nitrate (using UV-VIS) from the aqueous solutions. The structural changes of MLV precursors during green modification and the success of synthesis and biosorbent performances before and after the removal of anions from contaminated solutions have been registered and clarified by physicochemical methods, which include elemental analysis, chemical constitution, morphological characterization, and FTIR spectroscopy. In accordance with the FTIR spectroscopic analysis and isotherm studies, a structural model of the CALV biosorbent and its ion exchange mechanism are proposed.

Thermal Energy Storage of Composite Materials Based on Clay, Stearic Acid, Paraffin and Glauber’s Salt as Phase Change Materials

Thermal protection and insulation are important problems in many fields such as industry, agriculture and medicine. New composite materials with good thermal storage capacities have become important in the last few decades. The role of these materials is reflected in their ability to store energy and allow it to be reused in some other thermal systems. The aim of this study was to create a new material based on the basically activated bentonite clay. First, the clay was basically activated, resulting in a thick gel. Afterwards, stearic acid, Glauber’s salt and active carbon were added, and a heterogeneous gel was obtained as a finished final product. In order to obtain the best heterogeneous gel with satisfactory storage properties, the amount of stearic acid and Glauber’s salt was varied. The characterization of the resulting heterogeneous gel was performed by measuring the cooling rate of the gel samples. Compared with stearic acid, Glauber’s proved to be more effective. Heterogeneous gel cooling tests have shown that there was a certain proportional dependence between the concentration of stearic acid and the Glauber salt. However, it has been noticed the reduction in the cooling rate. Namely, the increase in stearic acid and Glauber’s salt concentration lead to slowing down the cooling rate of the gel. Adding active carbon to the heterogeneous gel also reduced the cooling rate, which indicated that the presence of active carbon in the heterogeneous gel should not be excluded in the future. The advantage of this system is the improvement of the gel thermal characteristics by the presence of water and clay. The gel was reversibly cooled and heated up to 100 °C without changing the homogeneous structure. This system can be used as a heat recovery pad, due to its flexible body pillow. It can be very quickly warmed up in a microwave oven if it is packaged in polyethylene packaging.

Scaling potential of geothermal water from the well A-2 at Vranjska Banja: Serbia

Geochemical and physicochemical characteristics of geothermal water from A-2 well are investigated in order to estimate its tendency towards the formation of scale in the pipe installation. Based on the geochemiocal analysis of major elements and groups, as well as triangular plots, this geothermal water can be classified as NaAlk·SO4 type. The ternary diagram classifies it as steamheated-peripheral water. A-2 has a good utilization capacity of 1 L s-1 at the water temperature of 84 °C. Regarding this, SI values are determined as the measure of tendency to form deposits. A positive SI value of calcite (0.14) and aragonite (0.03) at the temperature of 84 °C suggests a very low tendency to form scale, as well as the possible occurrence of a minor proportion of carbonate minerals. XRD mineralogical and SEM analyses of the scale have confirmed its calcite composition. SI values for the temperature of 25 °C indicate the safe silicate minerals deposition due to the positive temperature coefficient. The obtained SI values indicate that the A-2 geothermal water in Vranjska Banja is suitable for use in piping installations because it shows poorly pronounced tendency to form deposits.

Modification of quercetin with l-cysteine by horseradish peroxidase

Abstract Horseradish peroxidase is a well-known member of the peroxidase family that catalyzes oxidation of flavonoids and phenolic substrates to free phenoxyl or semiquinone radicals. Aim of this study was to investigate in vitro oxidation of quercetin by horseradish peroxidase in the presence of l-cysteine as nucleophilic agent, and its influence on previously formed semiquinone- and quinone-type metabolites. The obtained results showed that in the reaction without l-cysteine several products were present, such as quercetin quinone methide, phloroglucinol carboxylic acid, protocatechuic acid, as well as quercetin heterodimer and derivates of quercetin heterodimer. On the other hand, in the presence of l-cysteine only three products were obtained, quercetin quinone methide and two new isomeric mono-cysteine derivatives of quercetin with mass exp. m/z 420.04 ± 0.1 [quercetin + cysteine–H]– (theor. m/z 420.0389 [quercetin + cysteine–H]–).