Search results for: high perforamance liquid chromatography

Authors: Sameena Tarannum, S. Pranesh

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A nonlinear study of triple diffusive convection in a rotating couple stress liquid has been analysed. It is performed to study the effect of heat and mass transfer by deriving Ginzburg-Landau equation. Heat and mass transfer are quantified in terms of Nusselt number and Sherwood numbers, which are obtained as a function of thermal and solute Rayleigh numbers. The obtained Ginzburg-Landau equation is Bernoulli equation, and it has been elucidated numerically by using Mathematica. The effects of couple stress parameter, solute Rayleigh numbers, and Taylor number on the onset of convection and heat and mass transfer have been examined. It is found that the effects of couple stress parameter and Taylor number are to stabilize the system and to increase the heat and mass transfer.

Keywords: couple stress liquid, Ginzburg-Landau model, rotation, triple diffusive convection

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Authors: Moshe Mello, Hilary Rutto, Tumisang Seodigeng, Itumeleng Kohitlhetse

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The accelerating requirement to reach 0% sulfur content in liquid fuels demand researchers to seek efficient alternative technologies to challenge the predicament. In this current study, the adsorption capabilities of modified Cu(I)-Y zeolite were tested for the removal of organosulfur compounds (OSC) present in tire pyrolytic oil (TPO). The π-complexation-based adsorbent was obtained by ion exchanging Y-zeolite with Cu+ cation using liquid phase ion exchange (LPIE). Preparation of the adsorbent involved firstly ion exchange between Na-Y zeolite with a Cu(NO₃)₂ aqueous solution of 0.5M for 48 hours followed by reduction of Cu²⁺ to Cu+. Fixed-bed breakthrough studies for TPO in comparison with model diesel comprising of sulfur compounds such as thiophene, benzothiophenes (BT), and dibenzothiophenes (DBT) showed that modified Cu(I)-Y zeolite is an effective adsorbent for removal of OSC in liquid fuels. The effect of operating conditions such as adsorbent dosage and reaction time were studied to optimize the adsorptive desulfurization process. For model diesel fuel, the selectivity for adsorption of sulfur compounds followed the order DBT> BT> Thiophene. The Cu(I)-Y zeolite is fully regeneratable and this is achieved by a simple procedure of blowing the adsorbent with air at 350 °C, followed by reactivation at 450 °C in a rich helium surrounding.

Keywords: adsorption, desulfurization, TPO, zeolite

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Authors: Laure Wiest, Barbara Giroud, Azziz Assoumani, Francois Lestremau, Emmanuelle Vulliet

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Due to their production at high tonnages and their extensive use, surfactants are contaminants among those determined at the highest concentrations in wastewater. However, analytical methods and data regarding their occurrence in river water are scarce and concern only a few families, mainly anionic surfactants. The objective of this study was to develop an analytical method to extract and analyze a wide variety of surfactants in a minimum of steps, with a sensitivity compatible with the detection of ultra-traces in surface waters. 27 substances, from 12 families of surfactants, anionic, cationic and non-ionic were selected for method optimization. Different retention mechanisms for the extraction by solid phase extraction (SPE) were tested and compared in order to improve their detection by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). The best results were finally obtained with a C18 grafted silica LC column and a polymer cartridge with hydrophilic lipophilic balance (HLB), and the method developed allows the extraction of the three types of surfactants with satisfactory recoveries. The final analytical method comprised only one extraction and two LC injections. It was validated and applied for the quantification of surfactants in 36 river samples. The method's limits of quantification (LQ), intra- and inter-day precision and accuracy were evaluated, and good performances were obtained for the 27 substances. As these compounds have many areas of application, contaminations of instrument and method blanks were observed and considered for the determination of LQ. Nevertheless, with LQ between 15 and 485 ng/L, and accuracy of over 80%, this method was suitable for monitoring surfactants in surface waters. Application on French river samples revealed the presence of anionic, cationic and non-ionic surfactants with median concentrations ranging from 24 ng/L for octylphenol ethoxylates (OPEO) to 4.6 µg/L for linear alkylbenzenesulfonates (LAS). The analytical method developed in this work will therefore be useful for future monitoring of surfactants in waters. Moreover, this method, which shows good performances for anionic, non-ionic and cationic surfactants, may be easily adapted to other surfactants.

Keywords: anionic surfactant, cationic surfactant, LC-MS/MS, non-ionic surfactant, SPE, surface water

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Authors: Ruolan Zhang, Ryo Imai, Naoko Senda, Tomoyuki Sakai

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Enzymes have attracted increasing attentions in industrial manufacturing for their applicability in catalyzing complex chemical reactions under mild conditions. Directed evolution has become a powerful approach to optimize enzymes and exploit their full potentials under the circumstance of insufficient structure-function knowledge. With the incorporation of cell-free synthetic biotechnology, rapid enzyme synthesis can be realized because no cloning procedure such as transfection is needed. Its open environment also enables direct enzyme measurement. These properties of cell-free biotechnology lead to excellent throughput of enzymes generation. However, the capabilities of current screening methods have limitations. Fluorescence-based assay needs applicable fluorescent label, and the reliability of acquired enzymatic activity is influenced by fluorescent label’s binding affinity and photostability. To acquire the natural activity of an enzyme, another method is to combine pre-screening step and high-performance liquid chromatography (HPLC) measurement. But its throughput is limited by necessary time investment. Hundreds of variants are selected from libraries, and their enzymatic activities are then identified one by one by HPLC. The turn-around-time is 30 minutes for one sample by HPLC, which limits the acquirable enzyme improvement within reasonable time. To achieve the real high-throughput enzyme screening, i.e., obtain reliable enzyme improvement within reasonable time, a widely applicable high-throughput measurement of enzymatic reactions is highly demanded. Here, a high-throughput screening method using broadband coherent anti-Stokes Raman spectroscopy (CARS) was proposed. CARS is one of coherent Raman spectroscopy, which can identify label-free chemical components specifically from their inherent molecular vibration. These characteristic vibrational signals are generated from different vibrational modes of chemical bonds. With the broadband CARS, chemicals in one sample can be identified from their signals in one broadband CARS spectrum. Moreover, it can magnify the signal levels to several orders of magnitude greater than spontaneous Raman systems, and therefore has the potential to evaluate chemical's concentration rapidly. As a demonstration of screening with CARS, alcohol dehydrogenase, which converts ethanol and nicotinamide adenine dinucleotide oxidized form (NAD+) to acetaldehyde and nicotinamide adenine dinucleotide reduced form (NADH), was used. The signal of NADH at 1660 cm⁻¹, which is generated from nicotinamide in NADH, was utilized to measure the concentration of it. The evaluation time for CARS signal of NADH was determined to be as short as 0.33 seconds while having a system sensitivity of 2.5 mM. The time course of alcohol dehydrogenase reaction was successfully measured from increasing signal intensity of NADH. This measurement result of CARS was consistent with the result of a conventional method, UV-Vis. CARS is expected to have application in high-throughput enzyme screening and realize more reliable enzyme improvement within reasonable time.

Keywords: Coherent Anti-Stokes Raman Spectroscopy, CARS, directed evolution, enzyme screening, Raman spectroscopy

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Authors: Ifeanyichukwu Edeh, Tim Overton, Steve Bowra

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Currently biodiesel is produced from plant oils or animal’s fats by a liquid-phase catalysed transesterification process at low temperature. Although biodiesel is renewable and to a large extent sustainable, inherent properties such as poor cold flow, low oxidation stability, low cetane value restrict application to blends with fossil fuels. An alternative to biodiesel is renewable diesel produced by catalytic hydrotreating of oils and fats and is considered a drop in fuel because its properties are similar to petroleum diesel. In addition to developing alternative productions routes there is continued interest in reducing the cost of the feed stock, waste cooking oils and fats are increasingly used as the feedstocks due to low cost. However, use of oils and fat are highly adulterated resulting in high free fatty acid content which turn impacts on the efficiency of FAME production. Therefore, in light of the need to develop, alternative lipid feed stocks and related efficient catalysis the present study investigates the potential of producing renewable diesel from the lipids-extracted from activated sludge, a waste water treatment by-product, through catalytic hydrothermal decarboxylation. The microbial lipids were first extracted from the activated sludge using the Folch et al method before hydrothermal decarboxylation reactions were carried out using palladium (Pd/C) and platinum (Pt/C) on activated carbon as the catalysts in a batch reactor. The impact of three temperatures 290, 300, 330 °C and residence time between 30 min and 4hrs was assessed. At the end of the reaction, the products were recovered using organic solvents and characterized using gas chromatography (GC). The principle products of the reaction were pentadecane and heptadecane. The highest yields of pentadecane and heptadecane from lipid-extract were 23.23% and 15.21%, respectively. These yields were obtained at 290 °C and residence time 1h using Pt/C. To the best of our knowledge, the current work is the first investigation on the hydrothermal decarboxylation of lipid-extract from activated sludge.

Keywords: activated sludge, lipid, hydrothermal decarboxylation, renewable diesel

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Authors: Gargi Prasad, Ashiho A. Mao, Deepu Vijayan, S. Mandal

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The main objective of the present study was to optimize and develop an efficient protocol for in vitro propagation of a medicinally important orchid Cephalantheropsis obcordata (Lindl.) Ormerod along with genetic stability analysis of regenerated plants. This plant has been traditionally used in Chinese folk medicine and the decoction of whole plant is known to possess anticancer activity. Nodal segments used as explants were inoculated on Murashige and Skoog (MS) medium supplemented with various concentrations of isopentenyl adenine (2iP). The rooted plants were successfully acclimatized in the greenhouse with 100% survival rate. Inter-simple sequence repeats (ISSR) markers were used to assess the genetic fidelity of in vitro raised plants and the mother plant. It was revealed that monomorphic bands showing the absence of polymorphism in all in vitro raised plantlets analyzed, confirming the genetic uniformity among the regenerants. Phytochemical analysis was done to compare the antioxidant activities and HPLC fingerprinting assay of 80% aqueous ethanol extract of the leaves and stem of in vitro and in vivo grown C. obcordata. The extracts of the plants were examined for their antioxidant activities by using free radical 1, 1-diphenyl-2-picryl hydrazyl (DPPH) scavenging method, 2,2’-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging ability, reducing power capacity, estimation of total phenolic content, flavonoid content and flavonol content. A simplified method for the detection of ascorbic acid, phenolic acids and flavonoids content was also developed by using reversed phase high-performance liquid chromatography (HPLC). This is the first report on the micropropagation, genetic integrity study and quantitative phytochemical analysis of in vitro regenerated plants of C. obcordata.

Keywords: Cephalantheropsis obcordata, genetic fidelity, ISSR markers, HPLC

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Authors: Khadri Sihem, Abbaci Nafissa, Zerari Labiba

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In the context of the search for new bioactive natural products, we were interested in evaluating some antibacterial properties of two plant extracts: total phenols and flavonoids of Algerian medicinal plant. Our study occurs in two axes: The first concerns the extraction of phenolic compounds and flavonoids with methanol by liquid-liquid extraction, followed by quantification of the levels of these compounds in the end the analysis of the chemical composition of extracts. In the second axis, we studied the antibacterial power of the studied plant extracts.

Keywords: antibacterial activity, flavonoids, medicinal plants, polyphenols

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Authors: M. Y. Ismail, M. Inam

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This paper presents the design and analysis of Liquid Crystal (LC) based tunable reflect array antenna with different design configurations within X-band frequency range. The effect of LC volume used for unit cell element on frequency tunability and reflection loss performance has been investigated. Moreover different slot embedded patch element configurations have been proposed for LC based tunable reflect array antenna design with enhanced performance. The detailed fabrication and measurement procedure for different LC based unit cells has been presented. The waveguide scattering parameter measured results demonstrated that by using the circular slot embedded patch elements, the frequency tunability and dynamic phase range can be increased from 180 MHz to 200 MHz and 120° to 124° respectively. Furthermore the circular slot embedded patch element can be designed at 10 GHz resonant frequency with a patch volume of 2.71 mm3 as compared to 3.47 mm3 required for rectangular patch without slot.

Keywords: liquid crystal, tunable reflect array, frequency tunability, dynamic phase range

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Authors: Seyed Roozbeh Pishva, Alireza Aboudi Asl

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In this investigation a hydrocyclone by using for separation particles from fluid in oil and gas, mining and other industries is simulated. Case study is cone – cylindrical and solid - liquid hydrocyclone. The fluid is water and the solid is a type of silis having diameters of 53, 75, 106, 150, 212, 250, and 300 micron. In this investigation CFD method used for analysis flow and movement of particles in hydrocyclone. In this modeling flow is three-dimention, turbulence and RSM model have been used for solving. Particles are three dimensional, spherical and non rotating and for tracking them Lagrangian model is used. The results of this study in addition to analyzing flowfield, obtaining efficiency of hydrocyclone in 5, 7, 12, and 15 percent concentrations and compare them with experimental result that both of them had suitable agreement with each other.

Keywords: hydrocyclone, RSM Model, CFD, copper industry

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Authors: Hellal Abdelkader, Chafaa Salah, Touafri Lasnouni

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A simple, efficient and general method has been developed for the high diastereoselective synthesis of diethyl α-aminophosphonates in water through “one-pot” three-component reaction of aromatic aldehydes, aminophenols and dialkyl phosphites in the presence of a low catalytic amount (10mol%) of phosphorous acid as highly stable catalyst is described.

Keywords: DFT, HOMO-LUMO, phosphonic acid, aminophenols

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Authors: Leonardo Parra, Daniel Martínez, Jorge Pizarro, Fernando Ortega, Manuel Chacón-Fuentes, Andrés Quiroz

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Control of Dalaca pallens or blackworms, one of the most important hypogeous pest in grassland in southern Chile, is based on the use of broad-spectrum insecticides such as organophosphates and pyrethroids. However, the rapid development of insecticide resistance in field populations of this insect and public concern over the environmental impact of these insecticides has resulted in the search for other control methods. Specifically, the use of endophyte fungi for controlling pest has emerged as an interesting and promising strategy. Endophytes from ryegrass (Lolium perenne), establish a biotrophic relationship with the host, defined as mutualistic symbiosis. The plant-fungi association produces alkaloids where peramine is the main toxic substance against Listronotus bonariensis, the most important epigean pest of ryegrass. Nevertheless, the effect of peramina on others pest insects, such as D. pallens, to our knowledge has not been studied, and also its possible metabolization in the body of the larvae. Therefore, we addressed the following research question: Do larvae of D. pallens store peramine after consumption of ryegrass endophyte infected (E+)? For this, specimens of blackworms were fed with ryegrass plant of seven experimental lines and one commercial cultivar endophyte free (E-) sown at the Instituto de Investigaciones Agropecuarias Carillanca (Vilcún, Chile). Once the feeding period was over, ten larvae of each treatment were examined. Individuals were dissected, and their gut was removed to exclude any influence of remaining material. The rest of the larva's body was dried at 60°C by 24-48 h and ground into a fine powder using a mortar. 25 mg of dry powder was transferred to a microcentrifuge tube and extracted in 1 mL of a mixture of methanol:water:formic acid. Then, the samples were centrifuged at 16,000 rpm for 3 min, and the supernatant was colected and injected in the liquid chromatography of high resolution (HPLC). The results confirmed the presence of peramine in the larva's body of D. pallens. The insects that fed the experimental lines LQE-2 and LQE-6 were those where peramine was present in high proportion (0.205 and 0.199 ppm, respectively); while LQE-7 and LQE-3 obtained the lowest concentrations of the alkaloid (0.047 and 0.053 ppm, respectively). Peramine was not detected in the insects when the control cultivar Jumbo (E-) was tested. These results evidenced the storage and metabolism of peramine during consumption of the larvae. However, the effect of this alkaloid present in 'future ryegrass cultivars' (LQE-2 and LQE-6) on the performance and survival of blackworms must be studied and confirmed experimentally.

Keywords: blackworms, HPLC, alkaloid, pest

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Authors: Faezeh Mohammadi, Ebrahim Ebrahimi, Neda Azimi

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Use of ultrasound waves is one of the techniques for increasing the mixing and mass transfer in the microdevices. Ultrasound propagation into liquid medium leads to stimulation of the fluid, creates turbulence and so increases the mixing performance. In this study, CFD modeling of two-phase flow in a pitted micromixer equipped with a piezoelectric with frequency of 1.7 MHz has been studied. CFD modeling of micromixer at different velocity of fluid flow in the absence of ultrasound waves and with ultrasound application has been performed. The hydrodynamic of fluid flow and mixing efficiency for using ultrasound has been compared with the layout of no ultrasound application. The result of CFD modeling shows well agreements with the experimental results. The results showed that the flow pattern inside the micromixer in the absence of ultrasound waves is parallel, while when ultrasound has been applied, it is not parallel. In fact, propagation of ultrasound energy into the fluid flow in the studied micromixer changed the hydrodynamic and the forms of the flow pattern and caused to mixing enhancement. In general, from the CFD modeling results, it can be concluded that the applying ultrasound energy into the liquid medium causes an increase in the turbulences and mixing and consequently, improves the mass transfer rate within the micromixer.

Keywords: CFD modeling, ultrasound, mixing, mass transfer

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Authors: Li Feng, Cheng Zhang, Chuanzhou Yuang

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Understanding the macromolecular structure of low-rank coal is very important for its gasification and liquefaction. The pyrolysis is one of the methods of analyzing the macromolecular structure of coal. The gaseous products decomposed directly by the raw lignite at 500 °C and indirectly by tar products from raw lignite pyrolysis at 500 °C were investigated and compared by thermogravimetry, gas chromatography and mass spectrum coupled instrument system (TG/GC/MS) in this paper. The results show that 52 kinds of products were found from the raw lignite and 70 kinds of products from the tar. The pyrolysis products directly from the lignite appear more monocyclic aromatic hydrocarbons and less substituent groups or branch chain, compared with the products from the tar. There is less linear chain and double bonds structure in the tar, which can be speculated that linear chain and double bonds structure took part in the generation of condensed rings and other reactions. There are more kinds of phenol and furan in the tar, which indicate that these products may be generated from the secondary reaction. The formation process of phenol, phenol naphthalene, naphthene and furan are discussed.

Keywords: composition and structure, lignite, pyrolysis of coal, tar, TG/GC/MS

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Authors: Feiyu Yang, Chunfang Ni, Rong Wang, Yun Zou, Wenbin Liu, Chenggong Zhang, Fenjin Sun, Chun Wang

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Cocaine is the most frequently used illegal drug globally, with the global annual prevalence of cocaine used ranging from 0.3% to 0.4 % of the adult population aged 15–64 years. Growing consumption trend of abused cocaine and drug crimes are a great concern, therefore urine sample testing has become an important noninvasive sampling whereas cocaine and its metabolites (COCs) are usually present in high concentrations and relatively long detection windows. However, direct analysis of urine samples is not feasible because urine complex medium often causes low sensitivity and selectivity of the determination. On the other hand, presence of low doses of analytes in urine makes an extraction and pretreatment step important before determination. Especially, in gathered taking drug cases, the pretreatment step becomes more tedious and time-consuming. So developing a sensitive, rapid and high-throughput method for detection of COCs in human body is indispensable for law enforcement officers, treatment specialists and health officials. In this work, a new automated magnetic dispersive solid-phase extraction (MDSPE) sampling method followed by high performance liquid chromatography-mass spectrometry (HPLC-MS) was developed for quantitative enrichment of COCs from human urine, using prepared magnetic nanoparticles as absorbants. The nanoparticles were prepared by silanizing magnetic Fe3O4 nanoparticles and modifying them with divinyl benzene and vinyl pyrrolidone, which possesses the ability for specific adsorption of COCs. And this kind of magnetic particle facilitated the pretreatment steps by electromagnetically controlled extraction to achieve full automation. The proposed device significantly improved the sampling preparation efficiency with 32 samples in one batch within 40mins. Optimization of the preparation procedure for the magnetic nanoparticles was explored and the performances of magnetic nanoparticles were characterized by scanning electron microscopy, vibrating sample magnetometer and infrared spectra measurements. Several analytical experimental parameters were studied, including amount of particles, adsorption time, elution solvent, extraction and desorption kinetics, and the verification of the proposed method was accomplished. The limits of detection for the cocaine and cocaine metabolites were 0.09-1.1 ng·mL-1 with recoveries ranging from 75.1 to 105.7%. Compared to traditional sampling method, this method is time-saving and environmentally friendly. It was confirmed that the proposed automated method was a kind of highly effective way for the trace cocaine and cocaine metabolites analyses in human urine.

Keywords: automatic magnetic dispersive solid-phase extraction, cocaine detection, magnetic nanoparticles, urine sample testing

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Authors: Hela Ayeb Mrabtini, Ghazi Bellakhal, Jamel Chahed

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The presence of the dispersed phase in gas-liquid bubbly flow considerably alters the liquid turbulence. The bubbles induce turbulent fluctuations that enhance the global liquid turbulence level and alter the mechanisms of turbulence. RANS modeling of uniformly sheared flows on an isolated sphere centered in a control volume is performed using first and second order turbulence closures. The sphere is placed in the production-dissipation equilibrium zone where the liquid velocity is set equal to the relative velocity of the bubbles. The void fraction is determined by the ratio between the sphere volume and the control volume. The analysis of the turbulence statistics on the control volume provides numerical results that are interpreted with regard to the effect of the bubbles wakes on the turbulence structure in uniformly sheared bubbly flow. We assumed for this purpose that at low void fraction where there is no hydrodynamic interaction between the bubbles, the single-phase flow simulation on an isolated sphere is representative on statistical average of a sphere network. The numerical simulations were firstly validated against the experimental data of bubbly homogeneous turbulence with constant shear and then extended to produce numerical results for a wide range of shear rates from 0 to 10 s^-1. These results are compared with our turbulence closure proposed for gas-liquid bubbly flows. In this closure, the turbulent stress tensor in the liquid is split into a turbulent dissipative part produced by the gradient of the mean velocity which also contains the turbulence generated in the bubble wakes and a pseudo-turbulent non-dissipative part induced by the bubbles displacements. Each part is determined by a specific transport equation. The simulations of uniformly sheared flows on an isolated sphere reproduce the mechanisms related to the turbulent part, and the numerical results are in perfect accordance with the modeling of the transport equation of the turbulent part. The reduction of second order turbulence closure provides a description of the modification of turbulence structure by the bubbles presence using a dimensionless number expressed in terms of two-time scales characterizing the turbulence induced by the shear and that induced by bubbles displacements. The numerical simulations carried out in the framework of a comprehensive analysis reproduce particularly the attenuation of the turbulent friction showed in the experimental results of bubbly homogeneous turbulence subjected to a constant shear.

Keywords: gas-liquid bubbly flows, homogeneous turbulence, turbulence closure, uniform shear

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Authors: Pedro J. Rolim-Neto, Leslie R. M. Ferraz, Fabiana L. A. Santos, Pablo A. Ferreira, Ricardo T. L. Maia-Jr., Magaly A. M. Lyra, Danilo A F. Fonte, Salvana P. M. Costa, Amanda C. Q. M. Vieira, Larissa A. Rolim

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Initially developed as an antimalarial agent, hydroxychloroquine (HCQ) sulfate is often used as a slow-acting antirheumatic drug in the treatment of disorders of connective tissue. The United States Pharmacopeia (USP) 37 provides a reversed-phase HPLC method for quantification of HCQ. However, this method was not reproducible, producing asymmetric peaks in a long analysis time. The asymmetry of the peak may cause an incorrect calculation of the concentration of the sample. Furthermore, the analysis time is unacceptable, especially regarding the routine of a pharmaceutical industry. The aiming of this study was to develop a fast, easy and efficient method for quantification of HCQ sulfate by High Performance Liquid Chromatography (HPLC) based on the Quality by Design (QbD) methodology. This method was optimized in terms of peak symmetry using the surface area graphic as the Design of Experiments (DoE) and the tailing factor (TF) as an indicator to the Design Space (DS). The reference method used was that described at USP 37 to the quantification of the drug. For the optimized method, was proposed a 33 factorial design, based on the QbD concepts. The DS was created with the TF (in a range between 0.98 and 1.2) in order to demonstrate the ideal analytical conditions. Changes were made in the composition of the USP mobile-phase (USP-MP): USP-MP: Methanol (90:10 v/v, 80:20 v/v and 70:30 v/v), in the flow (0.8, 1.0 and 1.2 mL) and in the oven temperature (30, 35, and 40ºC). The USP method allowed the quantification of drug in a long time (40-50 minutes). In addition, the method uses a high flow rate (1,5 mL.min-1) which increases the consumption of expensive solvents HPLC grade. The main problem observed was the TF value (1,8) that would be accepted if the drug was not a racemic mixture, since the co-elution of the isomers can become an unreliable peak integration. Therefore, the optimization was suggested in order to reduce the analysis time, aiming a better peak resolution and TF. For the optimization method, by the analysis of the surface-response plot it was possible to confirm the ideal setting analytical condition: 45 °C, 0,8 mL.min-1 and 80:20 USP-MP: Methanol. The optimized HPLC method enabled the quantification of HCQ sulfate, with a peak of high resolution, showing a TF value of 1,17. This promotes good co-elution of isomers of the HCQ, ensuring an accurate quantification of the raw material as racemic mixture. This method also proved to be 18 times faster, approximately, compared to the reference method, using a lower flow rate, reducing even more the consumption of the solvents and, consequently, the analysis cost. Thus, an analytical method for the quantification of HCQ sulfate was optimized using QbD methodology. This method proved to be faster and more efficient than the USP method, regarding the retention time and, especially, the peak resolution. The higher resolution in the chromatogram peaks supports the implementation of the method for quantification of the drug as racemic mixture, not requiring the separation of isomers.

Keywords: analytical method, hydroxychloroquine sulfate, quality by design, surface area graphic

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Authors: Leonardo Parra, Manuel Chacón-Fuentes, Andrés Quiroz

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One of the most important factors in beef and dairy production in the world as well as also in Chile, is related to the correct choice of cultivars or mixtures of forage grasses and legumes to ensure high yields and quality of grassland. However, a great problem is the persistence of the grasses as a result of the action of different hypogeous as epigean pests. The complex insect pests associated with grassland include white grubs (Hylamorpha elegans, Phytoloema herrmanni), blackworm (Dalaca pallens) and Argentine stem weevil (Listronotus bonariensis). In Chile, the principal strategy utilized for controlling this pest is chemical control, through the use of synthetic insecticides, however, underground feeding habits of larval and flight activity of adults makes this uneconomic method. Furthermore, due to problems including environmental degradation, development of resistance and chemical residues, there is a worldwide interest in the use of alternative environmentally friendly pest control methods. In this sense, in recent years there has been an increasing interest in determining the role of endophyte fungi in controlling epigean and hypogeous pest. Endophytes from ryegrass (Lolium perenne), establish a biotrophic relationship with the host, defined as mutualistic symbiosis. The plant-fungi association produces a “cocktail of alkaloids” where peramine is the main toxic substance present in endophyte of ryegrass and responsible for damage reduction of L. bonariensis. In the last decade, few studies have been developed on the effectiveness of new ryegrass cultivars carriers of endophyte in controlling insect pests. Therefore, the aim of this research is to provide knowledge concerning to evaluate the alkaloid content, such as peramine and Lolitrem B, present in new experimental lines of ryegrass and feasible to be used in grasslands of southern Chile. For this, during 2016, ryegrass plants of six experimental lines and two commercial cultivars sown at the Instituto de Investigaciones Agropecuarias Carrillanca (Vilcún, Chile) were collected and subjected to a process of chemical extraction to identify and quantify the presence of peramine and lolitrem B by the technique of liquid chromatography of high resolution (HPLC). The results indicated that the experimental lines EL-1 and EL-3 had high content of peramine (0.25 and 0.43 ppm, respectively) than with lolitrem B (0.061 and 0.19 ppm, respectively). Furthermore, the higher contents of lolitrem B were detected in the EL-4 and commercial cultivar Alto (positive control) with 0.08 and 0.17 ppm, respectively. Peramine and lolitrem B were not detected in the cultivar Jumbo (negative control). These results suggest that EL-3 would have potential as future cultivate because it has high content of peramine, alkaloid responsible for controlling insect pest. However, their current role on the complex insects attacking ryegrass grasslands should be evaluated. The information obtained in this research could be used to improve control strategies against hypogeous and epigean pests of grassland in southern Chile and also to reduce the use of synthetic pesticides.

Keywords: HPLC, Lolitrem B, peramine, pest

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Authors: Y. A. Prada, Fanny Guzman, Rafael Cabanzo, John J. Castillo, Enrique Mejia-Ospino

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In this work, we show the important results about of synthesis of photoluminiscents gold nanoclusters using a small peptide as template for biosensing applications. Interestingly, we design one peptide (NBC2854) homologue to conservative domain from 215 250 residue of a galactolectin protein which can recognize the proteophosphoglycans (PPG) from Leishmania. Peptide was synthetized by multiple solid phase synthesis using FMoc group methodology in acid medium. Finally, the peptide was purified by High-Performance Liquid Chromatography using a Vydac C-18 preparative column and the detection was at 215 nm using a Photo Diode Array detector. Molecular mass of this peptide was confirmed by MALDI-TOF and to verify the α-helix structure we use Circular Dichroism. By means of the methodology used we obtained a novel fluorescents gold nanoclusters (AuNC) using NBC2854 as a template. In this work, we described an easy and fast microsonic method for the synthesis of AuNC with ≈ 3.0 nm of hydrodynamic size and photoemission at 630 nm. The presence of cysteine residue in the C-terminal of the peptide allows the formation of Au-S bond which confers stability to Peptide-based gold nanoclusters. Interactions between the peptide and gold nanoclusters were confirmed by X-ray Photoemission and Raman Spectroscopy. Notably, from the ultrafine spectra shown in the MALDI-TOF analysis which containing only 3-7 KDa species was assigned to Au₈-₁₈[NBC2854]₂ clusters. Finally, we evaluated the Peptide-gold nanocluster as an optical biosensor based on fluorescence spectroscopy and the fluorescence signal of PPG (0.1 µg-mL⁻¹ to 1000 µg-mL⁻¹) was amplified at the same wavelength emission (≈ 630 nm). This can suggest that there is a strong interaction between PPG and Pep@AuNC, therefore, the increase of the fluorescence intensity can be related to the association mechanism that take place when the target molecule is sensing by the Pep@AuNC conjugate. Further spectroscopic studies are necessary to evaluate the fluorescence mechanism involve in the sensing of the PPG by the Pep@AuNC. To our best knowledge the fabrication of an optical biosensor based on Pep@AuNC for sensing biomolecules such as Proteophosphoglycans which are secreted in abundance by parasites Leishmania.

Keywords: biosensing, fluorescence, Leishmania, peptide-gold nanoclusters, proteophosphoglycans

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Authors: Nader Nciri, Suil Song, Namho Kim, Namjun Cho

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Properties and compositions of bitumen and bitumen-derived liquids have significant influences on the selection of recovery, upgrading and refining processes. Optimal process conditions can often be directly related to these properties. The end uses of bitumen and bitumen products are thus related to their compositions. Because it is not possible to conduct a complete analysis of the molecular structure of bitumen, characterization must be made in other terms. The present paper focuses on physico-chemical analysis of two different types of bitumens. These bitumen samples were chosen based on: the original crude oil (sand oil and crude petroleum), and mode of process. The aim of this study is to determine both the manufacturing effect on chemical species and the chemical organization as a function of the type of bitumen sample. In order to obtain information on bitumen chemistry, elemental analysis (C, H, N, S, and O), heavy metal (Ni, V) concentrations, IATROSCAN chromatography (thin layer chromatography-flame ionization detection), FTIR spectroscopy, and 1H NMR spectroscopy have all been used. The characterization includes information about the major compound types (saturates, aromatics, resins and asphaltenes) which can be compared with similar data for other bitumens, more importantly, can be correlated with data from petroleum samples for which refining characteristics are known. Examination of Asphalt Ridge froth bitumen showed that it differed significantly from representative petroleum pitches, principally in their nonhydrocarbon content, heavy metal content and aromatic compounds. When possible, properties and composition were related to recovery and refining processes. This information is important because of the effects that composition has on recovery and processing reactions.

Keywords: froth bitumen, oil sand, asphalt ridge, petroleum pitch, thin layer chromatography-flame ionization detection, infrared spectroscopy, 1H nuclear magnetic resonance spectroscopy

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Authors: Z. Herceg, V. Stulic, T. Vukusic, A. Rezek Jambrak

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Gas phase plasma treatment is a new nonthermal technology used for food and water decontamination. In this study, we have investigated influence of the gas phase plasma treatment on yeast cells of S. cerevisiae. Sample was composed of 10 mL of yeast suspension and 190 mL of 0.01 M NaNO₃ with a medium conductivity of 100 µS/cm. Samples were treated in a glass reactor with a point- to-plate electrode configuration (high voltage electrode-titanium wire in the gas phase and grounded electrode in the liquid phase). Air or argon were injected into the headspace of the reactor at the gas flow of 5 L/min. Frequency of 60, 90 and 120 Hz, time of 5 and 10 min and positive polarity were defined parameters. Inactivation was higher with the applied higher frequency, longer treatment time and injected argon. Inactivation was not complete which resulted in complete recovery. Cellular leakage (260 nm and 280 nm) was higher with a longer treatment time and higher frequency. Leakage at 280 nm which defines a leakage of proteins was higher than leakage at 260 nm which defines a leakage of nucleic acids. The authors would like to acknowledge the support by Croatian Science Foundation and research project 'Application of electrical discharge plasma for preservation of liquid foods'.

Keywords: Saccharomyces cerevisiae, inactivation, gas-phase plasma treatment, cellular leakage

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Authors: Omasan Urhie Urhie, Adenipekun C. O, Eke W., Ogwu K., Erinle K. O

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Polychlorinated Biphenyls (PCBs) are Persistent organic pollutants (POPs) that are very toxic; they possess ability to accumulate in soil and in human tissues hence resulting in health issues like birth defect, reproductive disorder and cancer. The air is polluted by PCBs through volatilization and dispersion; they also contaminate soil and sediments and are not easily degraded. Soil samples were collected from a depth of 0-15 cm from three substations (Warri, Ughelli and Ibadan) of Power Holding Company of Nigeria (PHCN) where old transformers were dumped in Nigeria. Extraction and cleanup of soil samples were conducted using Accelerated Solvent Extraction (ASE) with Pressurized Liquid extraction (PLE). The concentration of PCBs was determined using gsas chromatography/mass spectrometry (GC/MS). Mean total PCB concentrations in the soil samples increased in the order Ughelli ˂ Ibadan˂ Warri, 2.457757ppm Ughelli substation 4.198926ppm, for Ibadan substation and 14.05065ppm at Warri substation. In the Warri samples, PCB-167 was the most abundant at about 30% (4.28086ppm) followed by PCB-157 at about 20% (2.77871), of the total PCB concentrations (14.05065ppm). Of the total PCBs in the Ughelli and Ibadan samples, PCB-156 was the most abundant at about 44% and 40%, respectively. This study provides a baseline report on the presence of PCBs in the vicinity of abandoned electrical power facilities in different cities in Nigeria.

Keywords: polychlorintated biphenyls, persistent organic pollutants, soil, transformer

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Authors: Filip Varga, Nina Jeran, Martina Biosic, Zlatko Satovic, Martina Grdisa

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Dalmatian pyrethrum (Tanacetum cinerariifolium /Trevir./ Sch. Bip.), a species endemic to the eastern Adriatic coastline, is the source of natural insecticide pyrethrin. Pyrethrin is a mixture of six compounds (pyrethrin I and II, cinerin I and II, jasmolin I and II) that exhibits high insecticidal activity with no detrimental effects to the environment. A recently optimized matrix-solid phase dispersion method (MSPD), using florisil as the sorbent, acetone-ethyl acetate (1:1, v/v) as the elution solvent, and sodium sulfate anhydrous as the drying agent was utilized to extract the pyrethrins from 10 wild populations (20 individuals per population) distributed along the Croatian coast. All six components in the extracts were qualitatively and quantitatively determined by high-performance liquid chromatography with a diode array detector (HPLC-DAD). Pearson’s correlation index was calculated between pyrethrin compounds, and differences between the populations using the analysis of variance were tested. Additionally, the correlation of each pyrethrin component with spatio-ecological variables (bioclimate, soil properties, elevation, solar radiation, and distance from the coastline) was calculated. Total pyrethrin content ranged from 0.10% to 1.35% of dry flower weight, averaging 0.58% across all individuals. Analysis of variance revealed significant differences between populations based on all six pyrethrin compounds and total pyrethrin content. On average, the lowest total pyrethrin content was found in the population from Pelješac peninsula (0.22% of dry flower weight) in which total pyrethrin content lower than 0.18% was detected in 55% of the individuals. The highest average total pyrethrin content was observed in the population from island Zlarin (0.87% of dry flower weight), in which total pyrethrin content higher than 1.00% was recorded in only 30% of the individuals. Pyrethrin I/pyrethrin II ratio as a measure of extract quality ranged from 0.21 (population from the island Čiovo) to 5.88 (population from island Mali Lošinj) with an average of 1.77 across all individuals. By far, the lowest quality of extracts was found in the population from Mt. Biokovo (pyrethrin I/II ratio lower than 0.72 in 40% of individuals) due to the high pyrethrin II content typical for this population. Pearson’s correlation index revealed a highly significant positive correlation between pyrethrin I content and total pyrethrin content and a strong negative correlation between pyrethrin I and pyrethrin II. The results of this research clearly indicate high intra- and interpopulation diversity of Dalmatian pyrethrum with regards to pyrethrin content and composition. The information obtained has potential use in plant genetic resources conservation and biodiversity monitoring. Possibly the largest potential lies in designing breeding programs aimed at increasing pyrethrin content in commercial breeding lines and reintroduction in agriculture in Croatia. Acknowledgment: This work has been fully supported by the Croatian Science Foundation under the project ‘Genetic background of Dalmatian pyrethrum (Tanacetum cinerariifolium /Trevir/ Sch. Bip.) insecticidal potential’ - (PyrDiv) (IP-06-2016-9034).

Keywords: Dalmatian pyrethrum, HPLC, MSPD, pyrethrin

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Authors: Suhas Kadam, Vishal Chandanshive, Niraj Rane, Sanjay Govindwar

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Fimbristylis dichotoma, Ammannia baccifera, and their co-plantation consortium FA were found to degrade methyl orange, simulated dye mixture, and real textile effluent. Wild plants of Fimbristylis dichotoma and Ammannia baccifera with equal biomass showed 91 and 89% decolorization of methyl orange within 60 h at a concentration of 50 ppm, while 95% dye removal was achieved by consortium FA within 48 h. Floating phyto-beds with co-plantation (Fimbristylis dichotoma and Ammannia baccifera) for the treatment of real textile effluent in a constructed wetland was observed to be more efficient and achieved 79, 72, 77, 66 and 56% reductions in ADMI color value, chemical oxygen demand, biological oxygen demand, total dissolve solid and total suspended solid of textile effluent, respectively. High performance thin layer chromatography, gas chromatography-mass spectroscopy, Fourier transform infrared spectroscopy, Ultra violet-Visible spectroscopy and enzymatic assays confirmed the phytotransformation of parent dye in the new metabolites. T-RFLP analysis of rhizospheric bacteria of Fimbristylis dichotoma, Ammannia baccifera, and consortium FA revealed the presence of 88, 98 and 223 genera which could have been involved in dye removal. Toxicity evaluation of products formed after phytotransformation of methyl orange by consortium FA on bivalves Lamellidens marginalis revealed less damage in the gills architecture when analyzed histologically. Toxicity measurement by Random Amplification of Polymorphic DNA (RAPD) technique revealed normal banding pattern in treated methyl orange sample suggesting less toxic nature of phytotransformed dye products.

Keywords: constructed wetland, phyto-bed, textile effluent, phytoremediation

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Authors: Antonella Di Sotto, Cinzia Ingallina, Caterina Fraschetti, Simone Circi, Marcello Locatelli, Simone Carradori, Gabriela Mazzanti, Luisa Mannina, Silvia Di Giacomo

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Cannabis sativa L. is one of the oldest cultivated plant species known not only for its voluptuous use but also for the wide application in food, textile, and therapeutic industries. Recently, the progress of biotechnologies applied to medicinal plants has allowed to produce different hemp varieties with low content of psychotropic phytoconstituents (tetrahydrocannabinol < 0.2% w/v), thus leading to a renewed industrial and therapeutic interest for this plant. In this context, in order to discover new potential remedies of pharmaceutical and/or nutraceutical interest, the chemopreventive properties of different organic and hydroalcoholic extracts, obtained from the inflorescences of C. sativa L. var. USO31, collected in June and September harvesting, were assessed. Particularly, the antimutagenic activity towards the oxidative DNA-damage induced by tert-butyl hydroperoxide (t-BOOH) was evaluated, and the DPPH (2,2-diphenyl-1-picrylhydrazyl) and ABTS (2,2'-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid) radical scavenging power of the samples were assessed as possible mechanisms of antimutagenicity. Furthermore, the ability of the extracts to inhibit the glucose-6-phosphate dehydrogenase (G6PD), whose overexpression has been found to play a critical role in neoplastic transformation and tumor progression, has been studied as a possible chemopreventive strategy. A careful phytochemical characterization of the extracts for phenolic and terpenoid composition has been obtained by high performance liquid chromatography (HPLC) and gas chromatography-mass spectrometry (GC-MS) methods. Under our experimental condition, all the extracts were found able to interfere with the tBOOH-induced mutagenicity in WP2uvrAR strain, although with different potency and effectiveness. The organic extracts from both the harvesting periods were found to be the main effective antimutagenic samples, reaching about a 55% inhibition of the tBOOH-mutagenicity at the highest concentration tested (250 μg/ml). All the extracts exhibited radical scavenger activity against DPPH and ABTS radicals, with a higher potency of the hydroalcoholic samples. The organic extracts were also able to inhibit the G6PD enzyme, being the samples from September harvesting the highly potent (about 50% inhibition respect to the vehicle). At the phytochemical analysis, all the extracts resulted to contain both polar and apolar phenolic compounds. The HPLC analysis revealed the presence of catechin and rutin as the major constituents of the hydroalcoholic extracts, with lower levels of quercetin and ferulic acid. The monoterpene carvacrol was found to be an ubiquitarian constituent. At GC-MS analysis, different terpenoids, among which caryophyllene sesquiterpenes, were identified. This evidence suggests a possible role of both polyphenols and terpenoids in the chemopreventive properties of the extracts from the inflorescences of C. sativa var. USO31. According to the literature, carvacrol and caryophyllene sesquiterpenes can contribute to the strong antimutagenicity although the role of all the hemp phytocomplex cannot be excluded. In conclusion, present results highlight a possible interest for the inflorescences of C. sativa var. USO31 as source of bioactive molecules and stimulate further studies in order to characterize its possible application for nutraceutical and pharmaceutical purposes.

Keywords: antimutagenicity, glucose-6-phosphate dehydrogenase, hemp inflorescences, nutraceuticals, sesquiterpenes

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Authors: Samiyah Tasleem, Muhammad Abdul Haq, Syed Baqir Shyum Naqvi, Muhammad Abid Husnain, Sajjad Haider Naqvi

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The objective of the present study was: a) to investigate the antimicrobial activity of honey samples of different floral sources of Pakistan, b) to recover the phenolic acids in them as a possible contributing factor of antimicrobial activity. Six honey samples from different floral sources, namely: Trachysperm copticum, Acacia species, Helianthus annuus, Carissa opaca, Zizyphus and Magnifera indica were used. The antimicrobial activity was investigated by the disc diffusion method against eight freshly isolated clinical isolates (Staphylococci aureus, Staphylococci epidermidis, Streptococcus faecalis, Pseudomonas aeruginosa, Klebsiella pneumonia, Escherichia coli, Proteus vulgaris and Candida albicans). Antimicrobial activity of honey was compared with five commercial antibiotics, namely: doxycycline (DO-30ug/mL), oxytetracycline (OT-30ug/mL), clarithromycin (CLR–15ug/mL), moxifloxacin (MXF-5ug/mL) and nystatin (NT – 100 UT). The fractions responsible for antimicrobial activity were extracted using ethyl acetate. Solid phase extraction (SPE) was used to recover the phenolic acids of honey samples. Identification was carried out via High-Performance Liquid Chromatography (HPLC). The results indicated that antimicrobial activity was present in all honey samples and found comparable to the antibiotics used in the study. In the microbiological assay, the ethyl acetate honey extract was found to exhibit a very promising antimicrobial activity against all the microorganisms tested, indicating the existence of phenolic compounds. Six phenolic acids, namely: gallic, caffeic, ferulic, vanillic, benzoic and cinnamic acids were identified besides some unknown substance by HPLC. In conclusion, Pakistani honey samples showed a broad spectrum antibacterial and promising antifungal activity. Identification of six different phenolic acids showed that Pakistani honey samples are rich sources of phenolic compounds that could be the contributing factor of antimicrobial activity.

Keywords: Pakistani honey, antimicrobial activity, Phenolic acids eg.gallic, caffeic, ferulic, vanillic, benzoic and cinnamic acids

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Authors: Dmitrii Glushkov, Aleksandr G. Nigay, Olga S. Yashutina

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In recent years, due to the intensive development of the Arctic and Antarctic areas, the actual task is to develop technology for the effective utilization of solid and liquid combustible wastes in an environment with low temperatures. Firstly, such technology will help to prevent the dumping of waste into the World Ocean and reduce the risks of causing environmental damage to the Far North areas. Secondly, promising actions will help to prepare fuel compositions from the waste in the places of their production. Such kind of fuels can be used as energy resources. It will reduce waste utilization costs when transporting them to the mainland. In the present study, we suggest a solution to the problem of waste utilization by the preparation of gel fuels based on solid and liquid combustible components with the addition of the thickener. Such kind of fuels is characterized by ease of preparation, storage, transportation and use (as energy resources). The main regularities and characteristics of physical and chemical processes are established with varying parameters of gel fuels and heating sources in wide ranges. The obtained results let us conclude about the prospects of gel fuels practical application for combustible wastes utilization. Appropriate technology will be characterized by positive environmental, operational and economic effects. The composition of the gel fuels can vary in a wide range. The fuels preparation based on one type of a combustible liquid or a several liquids mixture with the finely dispersed components addition makes it possible to obtain compositions with predicted rheological, energy or environmental characteristics. Besides, gel fuels have a lower level of the fire hazard compared to common solid and liquid fuels. This makes them convenient for storage and transportation. In such conditions, it is not necessary to transport combustible wastes from the territory of the Arctic and the Antarctic to the mainland for processing, which is now quite an expensive procedure. The research was funded by the Russian Science Foundation (project No. 18-13-00031).

Keywords: combustible liquid waste, gel fuel, ignition and combustion, utilization

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Authors: Siyabonga Aubrey Mhlongo, Linda Lunga Sibali, Phumlane Selby Mdluli, Peter Papoh Ndibewu, Kholofelo Clifford Malematja

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This paper reports on the successful extraction and quantification of an antibacterial and antifungal agent present in some consumer products (Triclosan: C₁₂H₇Cl₃O₂)generally found in wastewater or effluents using molecularly imprinted membrane adsorbent (MIMs) followed by quantification and removal on a high-performance liquid chromatography (HPLC). Triclosan is an antibacterial and antifungal agent present in some consumer products like toothpaste, soaps, detergents, toys, and surgical cleaning treatments. The MIMs was fabricated usingpolyvinylidene fluoride (PVDF) polymer with selective micro composite particles known as molecularly imprinted polymers (MIPs)via a phase inversion by immersion precipitation technique. This resulted in an improved hydrophilicity and mechanical behaviour of the membranes. Wastewater samples were collected from the Umbogintwini Industrial Complex (UIC) (south coast of Durban, KwaZulu-Natal in South Africa). central UIC effluent treatment plant and pre-treated before analysis. Experimental parameters such as sample size, contact time, stirring speed were optimised. The resultant MIMs had an adsorption efficiency of 97% of TCS with reference to NIMs and bare membrane, which had 92%, 88%, respectively. The analytical method utilized in this review had limits of detection (LoD) and limits of quantification (LoQ) of 0.22, 0.71µgL-1 in wastewater effluent, respectively. The percentage recovery for the effluent samples was 68%. The detection of TCS was monitored for 10 consecutive days, where optimum TCS traces detected in the treated wastewater was 55.0μg/L inday 9 of the monitored days, while the lowest detected was 6.0μg/L. As the concentrations of analytefound in effluent water samples were not so diverse, this study suggested that MIMs could be the best potential adsorbent for the development and continuous progress in membrane technologyand environmental sciences, lending its capability to desalination.

Keywords: molecularly imprinted membrane, triclosan, phase inversion, wastewater

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Authors: Nicholas Mavrogiannis, Francesca Crivellari, Zachary Gagnon

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Development of low-cost, rapid, sensitive and portable biosensing systems are important for the detection and prevention of disease in developing countries, biowarfare/antiterrorism applications, environmental monitoring, point-of-care diagnostic testing and for basic biological research. Currently, the most established commercially available and widespread assays for portable point of care detection and disease testing are paper-based dipstick and lateral flow test strips. These paper-based devices are often small, cheap and simple to operate. The last three decades in particular have seen an emergence in these assays in diagnostic settings for detection of pregnancy, HIV/AIDS, blood glucose, Influenza, urinary protein, cardiovascular disease, respiratory infections and blood chemistries. Such assays are widely available largely because they are inexpensive, lightweight, and portable, are simple to operate, and a few platforms are capable of multiplexed detection for a small number of sample targets. However, there is a critical need for sensitive, quantitative and multiplexed detection capabilities for point-of-care diagnostics and for the detection and prevention of disease in the developing world that cannot be satisfied by current state-of-the-art paper-based assays. For example, applications including the detection of cardiac and cancer biomarkers and biothreat applications require sensitive multiplexed detection of analytes in the nM and pM range, and cannot currently be satisfied with current inexpensive portable platforms due to their lack of sensitivity, quantitative capabilities and often unreliable performance. In this talk, inexpensive label-free biomolecular detection at liquid interfaces using a newly discovered electrokinetic phenomenon known as fluidic dielectrophoresis (fDEP) is demonstrated. The electrokinetic approach involves exploiting the electrical mismatches between two aqueous liquid streams forced to flow side-by-side in a microfluidic T-channel. In this system, one fluid stream is engineered to have a higher conductivity relative to its neighbor which has a higher permittivity. When a “low” frequency (< 1 MHz) alternating current (AC) electrical field is applied normal to this fluidic electrical interface the fluid stream with high conductivity displaces into the low conductive stream. Conversely, when a “high” frequency (20MHz) AC electric field is applied, the high permittivity stream deflects across the microfluidic channel. There is, however, a critical frequency sensitive to the electrical differences between each fluid phase – the fDEP crossover frequency – between these two events where no fluid deflection is observed, and the interface remains fixed when exposed to an external field. To perform biomolecular detection, two streams flow side-by-side in a microfluidic T-channel: one fluid stream with an analyte of choice and an adjacent stream with a specific receptor to the chosen target. The two fluid streams merge and the fDEP crossover frequency is measured at different axial positions down the resulting liquid

Keywords: biodetection, fluidic dielectrophoresis, interfacial polarization, liquid interface

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Authors: Song-Chol Pak, Yong-Jun Kim, Hak- Chol Choe, Yong-Son Choe

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We converted the coal-water-oil slurry (CWOS) into an alternative fuel (AF) for internal combustion engines by high-voltage discharge (HVD) and dielectric barrier discharge (DBD) plasmas. After its treatments, the CWOS had the average coal size reduced from 12.95 to 8.26㎛, improved dispersibility, fewer deposits, and calorific value enhanced by 35%. The effects of some parameters were analyzed on the conversion of CWOS to AF, and the AF was characterized. The plasma-treated CWOS is similar to other liquid fuels in rheological properties and calorific value. It is therefore concluded that it can be directly employed in internal combustion engines with a little design modification. The suggested method may be an alternative way of converting CWOS to AF without any dispersant or stabilizer.

Keywords: coal-water-oil slurry, high-voltage discharge, dielectric barrier discharge, plasma treatment, alternative fuel

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Authors: Vladimir Berezin, Andrey Bogoyavlenskiy, Pavel Alexyuk, Madina Alexyuk, Aizhan Turmagambetova, Irina Zaitseva, Nadezhda Sokolova, Elmira Omirtaeva

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Introduction: Triterpene saponins of plant origin are one of the most promising candidates for elaboration of novel adjuvants. Due to the combination of immunostimulating activity and the capacity interact with amphipathic molecules with formation of highly immunogenic nanocomplexes, triterpene saponins could serve as a good adjuvant/delivery system for vaccine use. In the research presented adjuvants on the base of nanocomplexes contained triterpene saponins isolated from Glycyrrhiza glabra and Saponaria officinalis plants indigenous to Kazakhstan were elaborated for influenza vaccine use. Methods: Purified triterpene saponins 'Glabilox' and 'SO1' with low toxicity and high immunostimulatory activity were isolated from plants Glycyrrhiza glabra L. and Saponaria officinalis L. by high-performance liquid chromatography (HPLC) and identified using electrospray ionization mass spectrometry (ESI-MS). Influenza virus A/St-Petersburg/5/09 (H1N1) propagated in 9-days old chicken embryos was concentrated and purified by centrifugation in sucrose gradient. Nanocomplexes contained lipids, and triterpene saponins Glabilox or SO1 were prepared by dialysis technique. Immunostimulating activity of experimental vaccine preparations was studied in vaccination/challenge experiments in mice. Results: Humoral and cellular immune responses and protection against influenza virus infection were examined after single subcutaneous and intranasal immunization. Mice were immunized subunit influenza vaccine (HA+NA) or whole virus inactivated influenza vaccine in doses 3.0/5.0/10.0 µg antigen/animal mixed with adjuvant in dose 15.0 µg/animal. Sera were taken 14-21 days following single immunization and mice challenged by A/St-Petersburg/5/09 influenza virus in dose 100 EID₅₀. Study of experimental influenza vaccine preparations in animal immunization experiments has shown that subcutaneous and intranasal immunization with subunit influenza vaccine mixed with nanocomplexes contained Glabilox or SO1 saponins stimulated high levels of humoral immune response (IgM, IgA, IgG1, IgG2a, and IgG2b antibody) and cellular immune response (IL-2, IL-4, IL-10, and IFN-γ cytokines) and resulted 80-90% protection against lethal influenza infection. Also, single intranasal and single subcutaneous immunization with whole virus inactivated influenza vaccine mixed with nanoparticulated adjuvants stimulated high levels of humoral and cellular immune responses and provided 100% protection against lethal influenza infection. Conclusion: The results of study have shown that nanocomplexes contained purified triterpene saponins Glabilox and SO1 isolated from plants indigenous to Kazakhstan can stimulate a broad spectrum of humoral and cellular immune responses and induce protection against lethal influenza infection. Both elaborated adjuvants are promising for incorporation to influenza vaccine intended for subcutaneous and intranasal routes of immunization.

Keywords: influenza vaccine, adjuvants, triterpene saponins, immunostimulating activity

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