Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 16

Search results for: microemulsion

16 Combination Approach Using Experiments and Optimal Experimental Design to Optimize Chemical Concentration in Alkali-Surfactant-Polymer Process

Authors: H. Tai Pham, Bae Wisup, Sungmin Jung, Ivan Efriza, Ratna Widyaningsih, Byung Un Min


The middle-phase-microemulsion in Alkaline-Surfactant-Polymer (ASP) solution and oil play important roles in the success of an ASP flooding process. The high quality microemulsion phase has ultralow interfacial tensions and it can increase oil recovery. The research used optimal experimental design and response-surface-methodology to predict the optimum concentration of chemicals in ASP solution for maximum microemulsion quality. Secondly, this optimal ASP formulation was implemented in core flooding test to investigate the effective injection volume. As the results, the optimum concentration of surfactants in the ASP solution is 0.57 wt.% and the highest effective injection volume is 19.33% pore volume.

Keywords: optimize, ASP, response surface methodology, solubilization ratio

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15 Formulation and Optimization of Topical 5-Fluorouracil Microemulsions Using Central Compisite Design

Authors: Sudhir Kumar, V. R. Sinha


Water in oil topical microemulsions of 5-FU were developed and optimized using face centered central composite design. Topical w/o microemulsion of 5-FU were prepared using sorbitan monooleate (Span 80), polysorbate 80 (Tween 80), with different oils such as oleic acid (OA), triacetin (TA), and isopropyl myristate (IPM). The ternary phase diagrams designated the microemulsion region and face centered central composite design helped in determining the effects of selected variables viz. type of oil, smix ratio and water concentration on responses like drug content, globule size and viscosity of microemulsions. The CCD design exhibited that the factors have statistically significant effects (p<0.01) on the selected responses. The actual responses showed excellent agreement with the predicted values as suggested by the CCD with lower residual standard error. Similarly, the optimized values were found within the range as predicted by the model. Furthermore, other characteristics of microemulsions like pH, conductivity were investigated. For the optimized microemulsion batch, ex-vivo skin flux, skin irritation and retention studies were performed and compared with marketed 5-FU formulation. In ex vivo skin permeation studies, higher skin retention of drug and minimal flux was achieved for optimized microemulsion batch then the marketed cream. Results confirmed the actual responses to be in agreement with predicted ones with least residual standard errors. Controlled release of drug was achieved for the optimized batch with higher skin retention of 5-FU, which can further be utilized for the treatment of many dermatological disorders.

Keywords: 5-FU, central composite design, microemulsion, ternanry phase diagram

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14 Dispersions of Carbon Black in Microemulsions

Authors: Mohamed Youssry, Dominique Guyomard, Bernard Lestriez


In order to enhance the energy and power densities of electrodes for energy storage systems, the formulation and processing of electrode slurries proved to be a critical issue in determining the electrode performance. In this study, we introduce novel approach to formulate carbon black slurries based on microemulsion and lyotropic liquid crystalline phases (namely, lamellar phase) composed of non-ionic surfactant (Triton X100), decanol and water. Simultaneous measurements of electrical properties of slurries under shear flow (rheology) have been conducted to elucidate the microstructure evolution with the surfactant concentration and decanol/water ratio at rest, as well as, the structural transition under steady-shear which has been confirmed by rheo-microscopy. Interestingly, the carbon black slurries at low decanol/water ratio are weak-gel (flowable) with higher electrical conductivity than those at higher ratio which behave strong-gel viscoelastic response. In addition, the slurries show recoverable electrical behaviour under shear flow in tandem with the viscosity trend. It is likely that oil-in-water microemulsion enhances slurries’ stability without affecting on the percolating network of carbon black. On the other hand, the oil-in-water analogous and bilayer structure of lamellar phase cause the slurries less conductive as a consequence of losing the network percolation. These findings are encouraging to formulate microemulsion-based electrodes for energy storage system (lithium-ion batteries).

Keywords: electrode slurries, microemulsion, microstructure transition, rheo-electrical properties

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13 Continuous Synthesis of Nickel Nanoparticles by Hydrazine Reduction

Authors: Yong-Su Jo, Seung-Min Yang, Seok Hong Min, Tae Kwon Ha


The synthesis of nickel nanoparticles by the reduction of nickel chloride with hydrazine in an aqueous solution. The effect of hydrazine concentration on batch-processed particle characteristics was investigated using Field Emission Scanning Electron Microscopy (FESEM). Both average particle size and geometric standard deviation (GSD) were decreasing with increasing hydrazine concentration. The continuous synthesis of nickel nanoparticles by microemulsion method was also studied using FESEM and X-ray Diffraction (XRD). The average size and geometric standard deviation of continuous-processed particles were 87.4 nm and 1.16, respectively. X-ray diffraction revealed continuous-processed particles were pure nickel crystalline with a face-centered cubic (fcc) structure.

Keywords: nanoparticle, hydrazine reduction, continuous process, microemulsion method

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12 Development of Biosurfactant-Based Adjuvant for Enhancing Biocontrol Efficiency

Authors: Kanyarat Sikhao, Nichakorn Khondee


Adjuvant is commonly mixed with agricultural spray solution during foliar application to improve the performance of microbial-based biological control, including better spreading, absorption, and penetration on a plant leaf. This research aims to replace chemical surfactants in adjuvant by biosurfactants for reducing a negative impact on antagonistic microorganisms and crops. Biosurfactant was produced from Brevibacterium casei NK8 and used as a cell-free broth solution containing a biosurfactant concentration of 3.7 g/L. The studies of microemulsion formation and phase behavior were applied to obtain the suitable composition of biosurfactant-based adjuvant, consisting of cell-free broth (70-80%), coconut oil-based fatty alcohol C12-14 (3) ethoxylate (1-7%), and sodium chloride (8-30%). The suitable formula, achieving Winsor Type III microemulsion (bicontinuous), was 80% of cell-free broth, 7% of fatty alcohol C12-14 (3) ethoxylate, and 8% sodium chloride. This formula reduced the contact angle of water on parafilm from 70 to 31 degrees. The non-phytotoxicity against plant seed of Oryza sativa and Brassica rapa subsp. pekinensis were obtained from biosurfactant-based adjuvant (germination index equal and above 80%), while sodium dodecyl sulfate and tween80 showed phytotoxic effects to these plant seeds. The survival of Bacillus subtilis in biosurfactant-based adjuvant was higher than sodium dodecyl sulfate and tween80. The mixing of biosurfactant and plant-based surfactant could be considered as a viable, safer, and acceptable alternative to chemical adjuvant for sustainable organic farming.

Keywords: biosurfactant, microemulsion, bio-adjuvant, antagonistic microorganisms

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11 Preparation and Characterization of Nickel-Tungsten Nanoparticles Using Microemulsion Mediated Synthesis

Authors: S. Pal, R. Singh, S. Sivakumar, D. Kunzru


AOT stabilized reverse micelles of deionized water, dispersed in isooctane have been used to synthesize bimetallic nickel tungsten nanoparticles. Prepared nanoparticles were supported on γ-Al2O3 followed by calcination at 500oC. Characterizations of the nanoparticles were done by TEM, XRD, FTIR, XRF, TGA and BET. XRF results showed that this method gave good composition control with W/Ni weight ratio equal to 3.2. TEM images showed particle size of 5-10 nm. Removal of surfactant after calcination was confirmed by TGA and FTIR.

Keywords: nanoparticles, reverse micelles, nickel, tungsten

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10 Improved Visible Light Activities for Degrading Pollutants on ZnO-TiO2 Nanocomposites Decorated with C and Fe Nanoparticles

Authors: Yuvraj S. Malghe, Atul B. Lavand


In recent years, semiconductor photocatalytic degradation processes have attracted a lot of attention and are used widely for the destruction of organic pollutants present in waste water. Among various semiconductors, titanium dioxide (TiO2) is the most popular photocatalyst due to its excellent chemical stability, non-toxicity, relatively low cost and high photo-oxidation power. It has been known that zinc oxide (ZnO) with band gap energy 3.2 eV is a suitable alternative to TiO2 due to its high quantum efficiency, however it corrodes in acidic medium. Unfortunately TiO2 and ZnO both are active only in UV light due to their wide band gaps. Sunlight consist about 5-7% UV light, 46% visible light and 47% infrared radiation. In order to utilize major portion of sunlight (visible spectrum), it is necessary to modify the band gap of TiO2 as well as ZnO. This can be done by several ways such as semiconductor coupling, doping the material with metals/non metals. Doping of TiO2 using transition metals like Fe, Co and non-metals such as N, C or S extends its absorption wavelengths from UV to visible region. In the present work, we have synthesized ZnO-TiO2 nanocomposite using reverse microemulsion method. Visible light photocatalytic activity of synthesized nanocomposite was investigated for degradation of aqueous solution of malachite green (MG). To increase the photocatalytic activity of ZnO-TiO2 nanocomposite, it is decorated with C and Fe. Pure, carbon (C) doped and carbon, iron(C, Fe) co-doped nanosized ZnO-TiO2 nanocomposites were synthesized using reverse microemulsion method. These composites were characterized using, X-ray diffraction (XRD), Energy dispersive X-ray spectroscopy (EDX), Scanning electron microscopy (SEM), UV visible spectrophotometery and X-ray photoelectron spectroscopy (XPS). Visible light photocatalytic activities of synthesized nanocomposites were investigated for degradation of aqueous malachite green (MG) solution. C, Fe co-doped ZnO-TiO2 nanocomposite exhibit better photocatalytic activity and showed threefold increase in photocatalytic activity. Effect of amount of catalyst, pH and concentration of MG solution on the photodegradation rate is studied. Stability and reusability of photocatalyst is also studied. C, Fe decorated ZnO-TiO2 nanocomposite shows threefold increase in photocatalytic activity.

Keywords: malachite green, nanocomposite, photocatalysis, titanium dioxide, zinc oxide

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9 Rheological Evaluation of a Mucoadhesive Precursor of Based-Poloxamer 407 or Polyethylenimine Liquid Crystal System for Buccal Administration

Authors: Jéssica Bernegossi, Lívia Nordi Dovigo, Marlus Chorilli


Mucoadhesive liquid crystalline systems are emerging how delivery systems for oral cavity. These systems are interesting since they facilitate the targeting of medicines and change the release enabling a reduction in the number of applications made by the patient. The buccal mucosa is permeable besides present a great blood supply and absence of first pass metabolism, it is a good route of administration. It was developed two systems liquid crystals utilizing as surfactant the ethyl alcohol ethoxylated and propoxylated (30%) as oil phase the oleic acid (60%), and the aqueous phase (10%) dispersion of polymer polyethylenimine (0.5%) or dispersion of polymer poloxamer 407 (16%), with the intention of applying the buccal mucosa. Initially, was performed for characterization of systems the conference by polarized light microscopy and rheological analysis. For the preparation of the systems the components described was added above in glass vials and shaken. Then, 30 and 100% artificial saliva were added to each prepared formulation so as to simulate the environment of the oral cavity. For the verification of the system structure, aliquots of the formulations were observed in glass slide and covered with a coverslip, examined in polarized light microscope (PLM) Axioskop - Zeizz® in 40x magnifier. The formulations were also evaluated for their rheological profile Rheometer TA Instruments®, which were obtained rheograms the selected systems employing fluency mode (flow) in temperature of 37ºC (98.6ºF). In PLM, it was observed that in formulations containing polyethylenimine and poloxamer 407 without the addition of artificial saliva was observed dark-field being indicative of microemulsion, this was also observed with the formulation that was increased with 30% of the artificial saliva. In the formulation that was increased with 100% simulated saliva was shown to be a system structure since it presented anisotropy with the presence of striae being indicative of hexagonal liquid crystalline mesophase system. Upon observation of rheograms, both systems without the addition of artificial saliva showed a Newtonian profile, after addition of 30% artificial saliva have been given a non-Newtonian behavior of the pseudoplastic-thixotropic type and after adding 100% of the saliva artificial proved plastic-thixotropic. Furthermore, it is clearly seen that the formulations containing poloxamer 407 have significantly larger (15-800 Pa) shear stress compared to those containing polyethyleneimine (5-50 Pa), indicating a greater plasticity of these. Thus, it is possible to observe that the addition of saliva was of interest to the system structure, starting from a microemulsion for a liquid crystal system, thereby also changing thereby its rheological behavior. The systems have promising characteristics as controlled release systems to the oral cavity, as it features good fluidity during its possible application and greater structuring of the system when it comes into contact with environmental saliva.

Keywords: liquid crystal system, poloxamer 407, polyethylenimine, rheology

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8 Microstructural Investigations of Metal Oxides Encapsulated Thermochromic Materials

Authors: Yusuf Emirov, Abdullatif Hakami, Prasanta K Biswas, Elias K Stefanakos, Sesha S Srinivasan


This study is aimed to develop microencapsulated thermochromic materials and the analysis of core-shell formation using high resolution electron microscopy. The candidate metal oxides (e.g., titanium oxide and silicon oxide) used for the microencapsulation of thermochromic materials are based on the microemulsion route that involves the micelle formation using different surfactants. The effectiveness of the core-shell microstructure formationrevealed the influence of surfactants and the metal oxide precursor concentrations. Additionally, a detailed thermal and color chromic behavior of these core-shell microcapsules are evaluated with the pristine thermochromic dye particles.

Keywords: core-shell thermochromic materials, core-shell microstructure formation, thermal and color chromic behavior of core-shell microcapsules, development micro-capsulated thermochromic materials

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7 Synthesis and Solubilization of Flurbiprofen Derivatives and Investigation of Their Biological Activities

Authors: Muhammad Mustaqeem, Musa Kaleem Baloch, Irfan Ullah, Ammarah Luqman, Afshan Ahmad


Flurbiprofen is one of the most potent nonsteroidal anti-inflammatory drugs. It is widely used for relief of pain in patients suffering from rheumatic diseases, migraine, sore throat and primary dysmenorrhea. However, its aqueous solubility is very low and hinders the skin permeation. Thus, it is imperative to develop such a drug delivery systems which can improve its aqueous solubility and hence improve the skin permeation and therapeutic compliance. Microemulsions have been also proven to increase the cutaneous absorption of lipophilic drugs as compared to conventional vehicles. Micro-emulsion is thermodynamically stable emulsion that has the capacity to ‘hide/solubilize’ water-insoluble molecules within a continuous oil phase. Therefore, flurbiprofen was converted to Easters through chemical reactions with alcohols such as methanol, ethanol, propanol and butanol. The product was further treated with hydrazine to get hydrazide. The solubility of the parent drug Flurbiprofen and the products were solubilized in microemulsions formed using various surfactants like ionic, non-ionic and zwitterions. It has been concluded that the product was more soluble than the parent compound. The biological activities of these were also investigated. The outcome was very promising and the product was more active than the parent compound. It, therefore, concluded that in this way, we can not only enhance the solubility of the drug and increase its bioactivity, but also reduce the risk of stomach cancer.

Keywords: Flurbiprofen, microemulsion, surfactants, hyrazides

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6 Magnetic Silica Nanoparticles as Viable Support for the Immobilization of Oxidative Enzymes

Authors: Y. Moldes-Diz, M. Gamallo, G. Eibes, C. Vazquez-Vazquez, G. Feijoo, J. M. Lema, M. T. Moreira


Laccases (benzenediol oxygen oxidoreductases, EC are excellent biocatalysts for biotechnological and environmental applications because of their high activity, selectivity, and specificity. Specifically, these characteristics allow them to perform the oxidation of recalcitrant compounds with simple requirements for the catalysis (presence of molecular oxygen). Nevertheless, the low stability under unfavorable conditions (pH, inactivating agents or temperature) and high production costs still limits their use for practical applications. Immobilization of enzymes has proven particularly valuable to avoid some of the aforementioned drawbacks. Magnetic nanoparticles (MNPs) have received increasing attention as carriers for enzyme immobilization since they can potentially provide an easy recovery of the biocatalyst from the reaction medium under an external magnetic field. In the present work, silica-coated magnetic nanoparticles ([email protected]) were prepared, characterized and used for laccase immobilization by covalent binding. The synthesis of [email protected] was performed in a two-step procedure: co-precipitation and reverse microemulsion. The influence of immobilization conditions: concentrations of the functionalization agent (3-aminopropyl-triethoxy-silane) and the cross-linker (glutaraldehyde) as well as the influence of pH, T or inactivating agents were evaluated. In general, immobilized laccase showed superior stability compared to that of free enzyme. The reusability of the biocatalyst was demonstrated in successive batch reactions, where enzyme activity was maintained above 65% after 8 cycles of oxidation of the substrate 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonate).

Keywords: silica-coated magnetic nanoparticles, laccase, immobilization, regeneration

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5 Effect of Surfactant Level of Microemulsions and Nanoemulsions on Cell Viability

Authors: Sonal Gupta, Rakhi Bansal, Javed Ali, Reema Gabrani, Shweta Dang


Nanoemulsions (NEs) and microemulsions (MEs) have been an attractive tool for encapsulation of both hydrophilic and lipophillic actives. Both these systems are composed of oil phase, surfactant, co-surfactant and aqueous phase. Depending upon the application and intended use, both oil-in-water and water-in-oil emulsions can be designed. NEs are fabricated using high energy methods employing less percentage of surfactant as compared to MEs which are self assembled drug delivery systems. Owing to the nanometric size of the droplets these systems have been widely used to enhance solubility and bioavailability of natural as well as synthetic molecules. The aim of the present study is to assess the effect of % age of surfactants on cell viability of Vero cells (African Green Monkeys’ Kidney epithelial cells) via MTT assay. Green tea catechin (Polyphenon 60) loaded ME employing low energy vortexing and NE employing high energy ultrasonication were prepared using same excipients (labrasol as oil, cremophor EL as surfactant and glycerol as co-surfactant) however, the % age of oil and surfactant needed to prepare the ME was higher as compared to NE. These formulations along with their excipients (oilME=13.3%, SmixME=26.67%; oilNE=10%, SmixNE=13.52%) were added to Vero cells for 24 hrs. The tetrazolium dye, 3-(4,5-dimethylthia/ol-2-yl)-2,5-diphi-iiyltclrazolium bromide (MTT), is reduced by live cells and this reaction is used as the end point to evaluate the cytoxicity level of a test formulation. Results of MTT assay indicated that oil at different percentages exhibited almost equal cell viability (oilME ≅ oilNE) while surfactant mixture had a significant difference in the cell viability values (SmixME < SmixNE). Polyphenon 60 loaded ME and its PlaceboME showed higher toxicity as compared to Polyphenon 60 loaded NE and its PlaceboNE that can be attributed to the higher concentration of surfactants present in MEs. Another probable reason for high % cell viability of Polyphenon 60 loaded NE might be due to the effective release of Polyphenon 60 from NE formulation that helps in the sustenance of Vero cells.

Keywords: cell viability, microemulsion, MTT, nanoemulsion, surfactants, ultrasonication

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4 Cationic Solid Lipid Nanoparticles Conjugated with Anti-Melantransferrin and Apolipoprotein E for Delivering Doxorubicin to U87MG Cells

Authors: Yung-Chih Kuo, Yung-I Lou


Cationic solid lipid nanoparticles (CSLNs) with anti-melanotransferrin (AMT) and apolipoprotein E (ApoE) were used to carry antimitotic doxorubicin (Dox) across the blood–brain barrier (BBB) for glioblastoma multiforme (GBM) treatment. Dox-loaded CSLNs were prepared in microemulsion, grafted covalently with AMT and ApoE, and applied to human brain microvascular endothelial cells (HBMECs), human astrocytes, and U87MG cells. Experimental results revealed that an increase in the weight percentage of stearyl amine (SA) from 0% to 20% increased the size of AMT-ApoE-Dox-CSLNs. In addition, an increase in the stirring rate from 150 rpm to 450 rpm decreased the size of AMT-ApoE-Dox-CSLNs. An increase in the weight percentage of SA from 0% to 20% enhanced the zeta potential of AMT-ApoE-Dox-CSLNs. Moreover, an increase in the stirring rate from 150 rpm to 450 rpm reduced the zeta potential of AMT-ApoE-Dox-CSLNs. AMT-ApoE-Dox-CSLNs exhibited a spheroid-like geometry, a minor irregular boundary deviating from spheroid, and a somewhat distorted surface with a few zigzags and sharp angles. The encapsulation efficiency of Dox in CSLNs decreased with increasing weight percentage of Dox and the order in the encapsulation efficiency of Dox was 10% SA > 20% SA > 0% SA. However, the reverse order was true for the release rate of Dox, suggesting that AMT-ApoE-Dox-CSLNs containing 10% SA had better-sustained release characteristics. An increase in the concentration of AMT from 2.5 to 7.5 μg/mL slightly decreased the grafting efficiency of AMT and an increase in that from 7.5 to 10 μg/mL significantly decreased the grafting efficiency. Furthermore, an increase in the concentration of ApoE from 2.5 to 5 μg/mL slightly reduced the grafting efficiency of ApoE and an increase in that from 5 to 10 μg/mL significantly reduced the grafting efficiency. Also, AMT-ApoE-Dox-CSLNs at 10 μg/mL of ApoE could slightly reduce the transendothelial electrical resistance (TEER) and increase the permeability of propidium iodide (PI). An incorporation of 10 μg/mL of ApoE could reduce the TEER and increase the permeability of PI. AMT-ApoE-Dox-CSLNs at 10 μg/mL of AMT and 5-10 μg/mL of ApoE could significantly enhance the permeability of Dox across the BBB. AMT-ApoE-Dox-CSLNs did not induce serious cytotoxicity to HBMECs. The viability of HBMECs was in the following order: AMT-ApoE-Dox-CSLNs = AMT-Dox-CSLNs = Dox-CSLNs > Dox. The order in the efficacy of inhibiting U87MG cells was AMT-ApoE-Dox-CSLNs > AMT-Dox-CSLNs > Dox-CSLNs > Dox. A surface modification of AMT and ApoE could promote the delivery of AMT-ApoE-Dox-CSLNs to cross the BBB via melanotransferrin and low density lipoprotein receptor. Thus, AMT-ApoE-Dox-CSLNs have appropriate physicochemical properties and can be a potential colloidal delivery system for brain tumor chemotherapy.

Keywords: anti-melanotransferrin, apolipoprotein E, cationic catanionic solid lipid nanoparticle, doxorubicin, U87MG cells

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3 Detailed Analysis of Mechanism of Crude Oil and Surfactant Emulsion

Authors: Riddhiman Sherlekar, Umang Paladia, Rachit Desai, Yash Patel


A number of surfactants which exhibit ultra-low interfacial tension and an excellent microemulsion phase behavior with crude oils of low to medium gravity are not sufficiently soluble at optimum salinity to produce stable aqueous solutions. Such solutions often show phase separation after a few days at reservoir temperature, which does not suffice the purpose and the time is short when compared to the residence time in a reservoir for a surfactant flood. The addition of polymer often exacerbates the problem although the poor stability of the surfactant at high salinity remains a pivotal issue. Surfactants such as SDS, Ctab with large hydrophobes produce lowest IFT, but are often not sufficiently water soluble at desired salinity. Hydrophilic co-solvents and/or co-surfactants are needed to make the surfactant-polymer solution stable at the desired salinity. This study focuses on contrasting the effect of addition of a co-solvent in stability of a surfactant –oil emulsion. The idea is to use a co-surfactant to increase stability of an emulsion. Stability of the emulsion is enhanced because of creation of micro-emulsion which is verified both visually and with the help of particle size analyzer at varying concentration of salinity, surfactant and co-surfactant. A lab-experimental method description is provided and the method is described in detail to permit readers to emulate all results. The stability of the oil-water emulsion is visualized with respect to time, temperature, salinity of the brine and concentration of the surfactant. Nonionic surfactant TX-100 when used as a co-surfactant increases the stability of the oil-water emulsion. The stability of the prepared emulsion is checked by observing the particle size distribution. For stable emulsion in volume% vs particle size curve, the peak should be obtained for particle size of 5-50 nm while for the unstable emulsion a bigger sized particles are observed. The UV-Visible spectroscopy is also used to visualize the fraction of oil that plays important role in the formation of micelles in stable emulsion. This is important as the study will help us to decide applicability of the surfactant based EOR method for a reservoir that contains a specific type of crude. The use of nonionic surfactant as a co-surfactant would also increase the efficiency of surfactant EOR. With the decline in oil discoveries during the last decades it is believed that EOR technologies will play a key role to meet the energy demand in years to come. Taking this into consideration, the work focuses on the optimization of the secondary recovery(Water flooding) with the help of surfactant and/or co-surfactants by creating desired conditions in the reservoir.

Keywords: co-surfactant, enhanced oil recovery, micro-emulsion, surfactant flooding

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2 Foslip Loaded and CEA-Affimer Functionalised Silica Nanoparticles for Fluorescent Imaging of Colorectal Cancer Cells

Authors: Yazan S. Khaled, Shazana Shamsuddin, Jim Tiernan, Mike McPherson, Thomas Hughes, Paul Millner, David G. Jayne


Introduction: There is a need for real-time imaging of colorectal cancer (CRC) to allow tailored surgery to the disease stage. Fluorescence guided laparoscopic imaging of primary colorectal cancer and the draining lymphatics would potentially bring stratified surgery into clinical practice and realign future CRC management to the needs of patients. Fluorescent nanoparticles can offer many advantages in terms of intra-operative imaging and therapy (theranostic) in comparison with traditional soluble reagents. Nanoparticles can be functionalised with diverse reagents and then targeted to the correct tissue using an antibody or Affimer (artificial binding protein). We aimed to develop and test fluorescent silica nanoparticles and targeted against CRC using an anti-carcinoembryonic antigen (CEA) Affimer (Aff). Methods: Anti-CEA and control Myoglobin Affimer binders were subcloned into the expressing vector pET11 followed by transformation into BL21 Star™ (DE3) E.coli. The expression of Affimer binders was induced using 0.1 mM isopropyl β-D-1-thiogalactopyranoside (IPTG). Cells were harvested, lysed and purified using nickle chelating affinity chromatography. The photosensitiser Foslip (soluble analogue of 5,10,15,20-Tetra(m-hydroxyphenyl) chlorin) was incorporated into the core of silica nanoparticles using water-in-oil microemulsion technique. Anti-CEA or control Affs were conjugated to silica nanoparticles surface using sulfosuccinimidyl-4-(N-maleimidomethyl) cyclohexane-1-carboxylate (sulfo SMCC) chemical linker. Binding of CEA-Aff or control nanoparticles to colorectal cancer cells (LoVo, LS174T and HC116) was quantified in vitro using confocal microscopy. Results: The molecular weights of the obtained band of Affimers were ~12.5KDa while the diameter of functionalised silica nanoparticles was ~80nm. CEA-Affimer targeted nanoparticles demonstrated 9.4, 5.8 and 2.5 fold greater fluorescence than control in, LoVo, LS174T and HCT116 cells respectively (p < 0.002) for the single slice analysis. A similar pattern of successful CEA-targeted fluorescence was observed in the maximum image projection analysis, with CEA-targeted nanoparticles demonstrating 4.1, 2.9 and 2.4 fold greater fluorescence than control particles in LoVo, LS174T, and HCT116 cells respectively (p < 0.0002). There was no significant difference in fluorescence for CEA-Affimer vs. CEA-Antibody targeted nanoparticles. Conclusion: We are the first to demonstrate that Foslip-doped silica nanoparticles conjugated to anti-CEA Affimers via SMCC allowed tumour cell-specific fluorescent targeting in vitro, and had shown sufficient promise to justify testing in an animal model of colorectal cancer. CEA-Affimer appears to be a suitable targeting molecule to replace CEA-Antibody. Targeted silica nanoparticles loaded with Foslip photosensitiser is now being optimised to drive photodynamic killing, via reactive oxygen generation.

Keywords: colorectal cancer, silica nanoparticles, Affimers, antibodies, imaging

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1 Impact of Electric Field on the Optical Properties of Hydrophilic Quantum Dots

Authors: Valentina V. Goftman, Vladislav A. Pankratov, Alexey V. Markin, Tangi Aubert, Zeger Hens, Sarah De Saeger, Irina Yu. Goryacheva


The most important requirements for biochemical applicability of quantum dots (QDs) are: 1) the surface cap should render intact or improved optical properties; 2) mono-dispersion and good stability in aqueous phase in a wide range of pH and ionic strength values; 3) presence of functional groups, available for bioconjugation; 4) minimal impact from the environment on the QDs’ properties and, vice versa, minimal influence of the QDs’ components on the environment; and 5) stability against chemical/biochemical/physical influence. The latter is especially important for in vitro and in vivo applications. For example, some physical intracellular delivery strategies (e.g., electroporation) imply a rapid high-voltage electric field impulse in order to temporarily generate hydrophilic pores in the cell plasma membrane, necessary for the passive transportation of QDs into the cell. In this regard, it is interesting to investigate how different capping layers, which can provide high stability and sufficient fluorescent properties of QDs in a water solution, behave under these abnormal conditions. In this contribution, hydrophobic core-shell CdSe/CdS/CdZnS/ZnS QDs (λem=600 nm), produced by means of the Successive Ion Layer Adsorption and Reaction (SILAR) technique, were transferred to a water solution using two of the most commonly used methods: (i) encapsulation in an amphiphilic brush polymer based on poly(maleic anhydride-alt-1-octadecene) (PMAO) modified with polyethylene glycol (PEG) chains and (ii) silica covering. Polymer encapsulation preserves the initial ligands on the QDs’ surface owing to the hydrophobic attraction between the hydrophobic groups of the amphiphilic molecules and the surface hydrophobic groups of the QDs. This covering process allows maintaining the initial fluorescent properties, but it leads to a considerable increase of the QDs’ size. However, covering with a silica shell, by means of the reverse microemulsion method, allows maintaining both size and fluorescent properties of the initial QDs. The obtained water solutions of polymer covered and silica-coated QDs in three different concentrations were exposed to a low-voltage electric field for a short time and the fluorescent properties were investigated. It is shown that the PMAO-PEG polymer acquires some additional charges in the presence of the electric field, which causes repulsion between the polymer and the QDs’ surface. This process destroys the homogeneity of the whole amphiphilic shell and it dramatically decreases the fluorescent properties (dropping to 10% from its initial value) because of the direct contact of the QDs with the strongly oxidative environment (water). In contrast, a silica shell possesses dielectric properties which allow retaining 90% of its initial fluorescence intensity, even after a longer electric impact. Thus, silica shells are clearly a preferable covering for bio-application of QDs, because – besides the high uniform morphology, controlled size and biocompatibility – it allows protecting QDs from oxidation, even under the influence of an electric field.

Keywords: electric field, polymer coating, quantum dots, silica covering, stability

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