Spontaneous Emulsification Research Articles

Essential oils of Lippia gracilis and Lippia sidoides chemotypes and their major compounds carvacrol and thymol: nanoemulsions and antifungal activity against Lasiodiplodia theobromae

The aim of this work was to evaluate the antifungal activity of essential oils (EOs) of Lippia gracilis and Lippia sidoides accessions, their major compounds and nanoemulsions. Nanoemulsions with 18% of EO or major compound were produced by spontaneous emulsification method. The EO of two L. gracilis accessions (LGRA-106 and LGRA-109) and two L. sidoides accessions (LSID-102 and LSID-104) were extracted by hydrodistillation and the major compounds thymol and carvacrol were purchased commercially. Antifungal activity was tested against Lasiodiplodia theobromae by calculating the percentage of inhibition of mycelial growth caused by different concentrations (0.1, 0.5, 1.0, 5.0, and 10.0 mL.L-1), in relation to the control. The EOs and their respective nanoemulsions presented as major compounds thymol (LGRA-106: 61.84% and Nano-106: 63.43%; LSID-102: 64.07% and Nano-102: 83.03%) or carvacrol (LGRA-109: 54.56% and Nano-109: 45.63%; LSID-104: 69.06% and Nano-104: 38.66%). Nano-104 presented 35.91% of an unidentified compound. The fungicidal activity of the EOs was similar to that of the major compounds, with a minimum fungicidal concentration of 1.0 mL.L-1 for LGRA-106, LSID-102 as well as for thymol, and 0.5 mL.L-1 for LGRA-109, LSID-104 as well as for carvacrol. The nanoemulsions were only able to reduce the mycelial growth of the fungus except for Nano-104 that exhibited fungicidal activity at the concentration of 10 ml.L-1. It was possible to observe that the EOs exhibited greater toxicity against L. theobromae than the nanoemulsions. These results can help in the development of products for the control of this important phytopathogen.

Interfacial Phenomena and Reaction Kinetics between High Al Molten Steel and CaO-SiO2-Type Flux

To understand the reaction mechanism between high Mn-high Al steel and slag, the reaction experiment of Fe-Mn-Al melts with CaO-SiO2-type flux was carried out in MgO crucible at 1873 K. The evolution of the morphology of interface was inspected firstly, and then the global reaction kinetics was modeled in consideration of the effect of dynamic interfacial phenomena. The results show that in the reaction of Fe-5 mass % Al alloy with high SiO2 or low SiO2 protective slag, the strong chemical affinity between the metal and flux leads to strong spontaneous emulsification and attenuated with the progression of the reaction. Combined with the change of interfacial area caused by emulsification, it is found that the global reaction kinetics can be described satisfactorily by the mass transfer model of Al in liquid steel, and the determined mass transfer coefficient was about k[Al]=4.46×10−5 m/s. However, the emulsification phenomenon in the reaction of Fe-13%Mn-5%Al alloy with low SiO2 slag did not disappear with the reaction, which can be attributed to the decreasing of the interfacial tension with Mn addition and the accumulation of C on the interface. This reaction process can be modeled by assuming the mass transfer of SiO2 in the slag as the rate-controlling step with the estimated transfer coefficient of k(SiO2)=5.12×10−6 m/s.

Direct Insight into the Cause and Time Evaluation of Spontaneous Emulsification of Water in Crude Oil during Low Salinity Waterflooding Using Microfluidic Model

Direct Insight into the Cause and Time Evaluation of Spontaneous Emulsification of Water in Crude Oil during Low Salinity Waterflooding Using Microfluidic Model

Ouzo polymerization: A bottom-up green synthesis of polymer nanoparticles by free-radical polymerization of monomers spontaneously nucleated by the Ouzo effect; Application to molecular imprinting

HypothesisTop-down fabrication of polymer nanoparticles from preformed polymers by spontaneous colloid formation under influence of the Ouzo effect is a widely applied concept whereas bottom-up free-radical polymerization of monomers nucleated under the Ouzo regime have found limited application after a seminal report almost half a century ago. We hypothesized that the approach would be of value today as a versatile method for green synthesis of polymer nanoparticles, including molecularly imprinted ones. ExperimentsTernary mixtures of pentaerythritol triacrylate, ethanol, and water were prepared by either a one-pot-one-step batch-wise procedure or a continuous-flow process using a 3D-printed micro-fluidic mixer. The mixtures were subjected to free-radical polymerization. Cortisol was added as a template to generate a molecular memory through molecular imprinting. Characterization of the fabricated nanoparticles was carried out by dynamic light scattering, electron microscopy, and binding studies. FindingsCompositions in the Ouzo region of the ternary phase diagram provided spontaneous and instantaneous formation of nucleated monomer droplets. Free-radical polymerization, promoted by heat or UV light, transformed the droplets into polymer nanoparticles in a green and sustainable route. Addition of cortisol created polymer nanoparticles with high affinity for cortisol in aqueous media. Competition studies showed some cross-reaction with other steroids, comparable to that found with antibodies, and complete discrimination of structurally unrelated drug molecules.

Influence of the Dissolved Gas on the Interfacial Properties of Decane Surface Nanodroplets.

Surface nanodroplets have received extensive attention recently due to their potential in the fabrication of functional materials with nanostructures and chemical reactions at micro- and nanoscales. Although the effect of dissolved gas in water has been realized in some important processes such as spontaneous emulsification of oil droplets in water, its roles in the wetting behavior of surface nanodroplets at the hydrophobic interface have been largely neglected. Here, we focused on the influence of dissolved gas on the interfacial properties of surface nanodroplets and characterized their morphological evolution when exposed to different air-saturated water samples. Results indicated that the morphology of surface nanodroplets barely changed in air-oversaturated cold water. However, their contact angle first decreases gradually in deionized water, increases immediately after replacement with degassed water, and eventually decreases gradually with time. Furthermore, the surface tension of nanodroplets would change similarly after the injection of degassed water. We considered these changes to be caused by the removal or reduction of the enriched gas at the substrate interface, in which the surface hydrophobicity was changed. Our findings could shed some light on the wetting behavior of nanodroplets at the hydrophobic surface in different air-saturated water samples and inspire the microscale manipulation and reaction of surface nanodroplets.

Phytochemical Analysis Both of Water and Ethanol Extract from Some Herbs Combinations, Nanoemulsion Formulation, and Antioxidant Effects

AIM: The purpose of this study was to analyze the phenolic content and antioxidant activity of both water and ethanol extracts of a combination of several herbs (Phyllanthus urinaria, Andrographis paniculata, Curcuma longa, Zingiber officinale, Citrus limon, and Cymbopogon citratus), as well as product development in the form of nanoemulsions. METHODS: The research was conducted by making three combinations of herbs mixtures with various compositions (Formulas A, B, and C). The combined herbs powder was then made in the form of water and ethanol extracts. The ethanol extract of each herbal combination was then made also in the form of a nanoemulsion by spontaneous emulsion. RESULTS: The ethanol extract of each herbs combination contained higher phenolic compounds and antioxidant activity than the water extract. The nanoemulsion product of the combined herbs ethanol extract had a particle size of 21.1; 34.9; and 50.7 nm and showed high antioxidant activity. CONCLUSION: The combined several herbs (P. urinaria, A. paniculata, C. longa, Z. officinale, C. limon, and C. citratus) ethanol extract and its nanoemulsion products have the potential to be developed as natural antioxidants.

Physicochemical properties and stability of nanoemulsions containing Clinacanthus nutans extract for postherpetic neuralgia

The leaf extract of Clinacanthus nutans (CNE), a medicinal plant widely distributed in South East Asia, has long been used in traditional medicine by applying topically for treatment of herpes zoster. Besides, it has an analgesic effects that could help relieve suffering from postherpetic neuralgia. The aims of this study were to develop nanoemulsions containing CNE obtained from extraction of dried leaves and evaluate their physicochemical properties, stability, and permeability. Forty formulations of oil-in-water emulsion bases containing various concentrations of sunflower oil and mixed emulsifiers were prepared by spontaneous emulsification and screened for further preparation of nanoemulsion bases by high presure homogenization. The results showed that particle sizes and viscosities of nanoemulsion bases were in the range of 85.9 – 120.9 nm and 339.4 – 1,362.0 cP, respectively. The content of vitexin, which was used as a marker in this study, in CNE and 4 %w/w CNE nanoemulsions were 0.130 and 0.005 %w/w, respectively. The 4 %w/w CNE nanoemulsions was found to be stable based on vitexin content under storage at 5 °C and 30 °C. The amount permeated of vitexin after 24-h permeation from 4 %w/w CNE nanoemulsion across Strat-M® membrane was 61.1 ± 1.95%. In conclusion, the promising formulation of nanoemulsions according to stability and permeability results was composed of 4 %w/w CNE, 20 %w/w sun flower oil, and 20 %w/w mixed emulsifiers of Span®80 and Brij®721 (weight ratio of 1.0:0.32), which could be a candidate as a herbal topical dosage form for postherpetic neuralgia.

Emulsification/demulsification method coupled to GC–MS/MS for analysis of multiclass pesticide residues in edible oils

A simple, rapid, and sensitive method was developed for simultaneous determination of 103 multiclass pesticides in edible oils. A new strategy of sample preparation involving a spontaneous emulsification followed by membrane-based demulsification was proposed. The developed strategy was compared with other reputed clean-up procedures and found to improve the removal efficiency of matrix interferences without a significant loss of pesticides. GC–MS/MS was used to evaluate the proposed treatment methodologies. Recovery studies were performed at 10, 20, and 50 μg/kg levels in soybean oil, olive oil, and rapeseed oil. Throughout the validations, 70–120% overall recoveries were achieved with RSDs < 10% for the majority of pesticides. Limits of quantification ranged from 10 to 50 μg/kg (10 μg/kg for over 92% pesticides), less than the regulatory MRLs. Twenty-two of 30 market samples were found to contain one or several detectable pesticide residues ranged from < 1 to 105.9 μg/kg.

Propylene Glycol Caprylate-Based Nanoemulsion Formulation of Plumbagin: Development and Characterization of Anticancer Activity.

Plumbagin, a bioactive naphthoquinone, has demonstrated potent antitumor potential. However, plumbagin is a sparingly water-soluble compound; therefore, clinical translation requires and will be facilitated by the development of a new pharmaceutical formulation. We have generated an oil-in-water nanoemulsion formulation of plumbagin using a low-energy spontaneous emulsification process with propylene glycol caprylate (Capryol 90) as an oil phase and Labrasol/Kolliphor RH40 as surfactant and cosurfactant excipients. Formulation studies using Capryol 90/Labrasol/Kolliphor RH40 components, based on pseudoternary diagram and analysis of particle size distribution and polydispersity determined by dynamic light scattering (DLS), identified an optimized composition of excipients for nanoparticle formulation. The nanoemulsion loaded with plumbagin as an active pharmaceutical ingredient had an average hydrodynamic diameter of 30.9 nm with narrow polydispersity. The nanoemulsion exhibited long-term stability, as well as good retention of particle size in simulated physiological environments. Furthermore, plumbagin-loaded nanoemulsion showed an augmented cytotoxicity against prostate cancer cells PTEN-P2 in comparison to free drug. In conclusion, we generated a formulation of plumbagin with high loading drug capacity, robust stability, and scalable production. Novel Capryol 90-based nanoemulsion formulation of plumbagin demonstrated antiproliferative activity against prostate cancer cells, warranting thus further pharmaceutical development.

Interfacial reaction-induced roughening in polymer thin films.

Reactive blending of immiscible polymers is an important process for synthesizing polymer blends with superior properties. We use a phase-field model to understand reaction dynamics and morphology evolution by diffusive transport in layered films of incompatible, end-reactive polymers. We thoroughly investigate this phenomenon over a large parameter space of interface shapes, layer thicknesses, reaction rates specified by a Damkohler number (Daf), and Flory-Huggins interaction parameter (χ), under static conditions with no external fields. For films of the same thickness, the dynamics of the system is not significantly influenced by the length of the film or the initial shape of the interface. The interface between the polymers is observed to roughen, leading to the formation of a spontaneous emulsion. The reaction progresses slower and the interface roughens later for thicker films, and systems with higher χ. Increasing Daf increases the reaction rate and hastens the onset of roughening. The quasi-static interfacial tension decreases with the extent of reaction, but does not become vanishingly small or negative at the onset of roughening. Simulations with reversible reactions and systems where only a fraction of the homopolymers have reactive end groups show that a critical diblock (reaction product) concentration exists, below which interfacial roughening and spontaneous emulsification is not observed. We also demonstrate that thermal fluctuations accelerate the onset of interfacial roughening, and help sustain the system in an emulsified state.

Nanoemulsification synthesis route for obtaining highly efficient Ag3PO4 photocatalytic nanomaterial

Nanoemulsion technique based on Ouzo effect was applied for the fast and simple synthesis of Ag3PO4 at room temperature. X-ray powder diffraction analysis and Raman spectroscopy reviled that synthesized powder was single-phase. Using scanning electron microscopy analysis, it was found that the synthesized Ag3PO4 particles were near-spherical shape with an average diameter of 100 nm. The high value for the specific surface area of obtained powder was measured by Brunauer?Emmet?Teller method. Finally, the Ag3PO4 product was used as a photocatalyst for the photodegradation of crystal violet dye in an aqueous solution. Nanoemulsion strategy procedure provides a simple pathway to obtain a highly efficient single-phase Ag3PO4 photocatalyst.

Development and in vivo evaluation of lipid-based nanocarriers containing Jatropha isabellei dry extract from the dichloromethane fraction intended for oral treatment of arthritic diseases

In this study, a dichloromethane fraction dry extract from the underground parts of Jatropha isabellei (DFJi) was used to prepare lipid nanocarriers (LNCJi) aimed at providing the oral delivery of terpenic compounds in the treatment of arthritis. The lipid nanocarriers were prepared by the spontaneous emulsification method. The lipid nanocarriers displayed sizes ranging from 180 to 200 nm and zeta potential values of around -18 mV. A high value of entrapment efficiency (> 90%) was obtained for jatrophone, which was used as the chemical marker of DFJi. LNCJi stored at 4°C were demonstrated to be stable through measurements of transmitted light after analytical centrifugation of the samples. In vitro drug release studies conducted in biorelevant dissolution media demonstrated that jatrophone release was faster from LNCJi than from free DFJi. When tested in an acute arthritis model, the LNCJi exhibited antinociceptive properties after oral administration of a 50 mg/kg dose, unlike the free DFJi, although no reduction in articular diameter was observed. These results suggest that an increase in the oral absorption of DFJi constituents may have occurred through the carrying of this fraction in LNCJi, thus improving the antinociceptive activity of this compound.

Preparation and interfacial behavior of surface-active microspheres for both emulsion stabilization and profile control

Polymer microspheres have high swelling ratio, which can be used to improve the heterogeneity of low permeability reservoirs. When combined with a spontaneous emulsifying flooding system, the sweep efficiency of the system can be increased. However, due to its strong hydrophilicity, the conventional polymer microspheres cannot be adsorbed on the oil-water interface and reduce the interfacial tension, so as to improve the stability of emulsion and further improve the oil washing efficiency of spontaneous emulsion flooding. In this paper, surface-active microspheres S-PM are developed by using stearyl methacrylate (SMA) as the surface-active monomer. Compared with conventional polymer microspheres, the microspheres are amphiphilic particles with a swelling particle size of 600 nm, which can improve the stability of emulsions while improving the heterogeneity of low permeability reservoirs. The study of interfacial behavior shows that S-PM can decrease the interfacial tension to 0.005 mN/m by dragging surfactant molecules to the oil-water interface through polar action and increasing the surfactant concentration on the interface, and can increase the interfacial dilatational elastic modulus to 26.5 mN/m by forming a rigid adsorption layer. The emulsion stability analysis and micrographs show that S-PM can be distributed on the surface of the droplet. Due to the decrease of oil-water interfacial tension and the increase of interfacial strength, it is not easy to collide and coalesce, thus improving the emulsion stability. The dewatering rate decreases from 81.6% to 65.4%, and the TSI decreases from 9.7 to 5.3. The profile control results show that after adding S-PM, the spontaneous emulsifying flooding system is suitable for heterogeneous low-permeability reservoirs with permeability ratio below 5. This study expands the function of polymer microspheres, and it is expected to further improve the recovery of low permeability reservoirs by combining surface-active microspheres with a spontaneous emulsion flooding system.

CURCUMINOID NANOEMULSION FROM Curcuma xanthorrhiza EXTRACT AND ITS ACTIVITY AS ANTIOXIDANT, ANTIBACTERIAL AND ANTIFUNGAL

Curcuminoids are the main compounds in the rhizome extract of C. xanthorrhiza which are beneficial for health. The aim of the research was to study the formulation of curcuminoid nanoemulsions and their biological effects as antioxidant, antibacterial, and antifungal agents. The formulation of the curcuminoid nanoemulsion was carried out using the spontaneous emulsion method, by mixing the curcuminoid extract with oil and tween 80 and adding phosphate buffer pH 7.0. Curcuminoid nanoemulsion products were characterized and tested for their potential biological activity. Antioxidant test using reagent 2,2-diphenyl-1-picrylhydrazyl. Antibacterial test using diffusion method against Escherichia coli and Staphylococcus epidermidis bacteria. Antifungal test using Candida albicans. The curcuminoid nanoemulsion product showed a particle size of 19.5 to 32.1 nm, a polydispersity index &lt; 0.3, and a zeta potential of -4.5 to -9.2 mV. Curcuminoid nanoemulsions have high antioxidant and antifungal activity, while antibacterial activity showed moderate activity. The conclusion is that curcuminoid nanoemulsions from C. xanthorrhiza extract can be used effectively as antioxidants and antimicrobial for commercial applications.

Janus Photonic Microspheres with Bridged Lamellar Structures via Droplet-Confined Block Copolymer Co-Assembly.

Artificial self-assembly systems typically exhibit limited capability in creating nature-inspired complex materials with advanced functionalities. Here, an effective co-assembly strategy is demonstrated for the facile creation of complex photonic structures with intriguing light reflections. Two different lipophilic and amphiphilic bottlebrush block copolymers (BCPs) are placed within shrinking droplets to enable a cooperative working mechanism of microphase segregation and organized spontaneous emulsification, respectively. Layer assemblies of the lipophilic BCP and uniform water nanodroplets stabilized by the bottlebrush surfactant are both generated, and co-assembled into a bridged lamellar structure with the alternating arrangement of layers and closely packed nanodroplet arrays. Janus microspheres with diverse dual optical characteristics are successfully fabricated, and reflected wavelengths of light are highly tunable simply by changing the formulation or molecular weight of BCP.

Janus Photonic Microspheres with Bridged Lamellar Structures via Droplet‐Confined Block Copolymer Co‐Assembly

AbstractArtificial self‐assembly systems typically exhibit limited capability in creating nature‐inspired complex materials with advanced functionalities. Here, an effective co‐assembly strategy is demonstrated for the facile creation of complex photonic structures with intriguing light reflections. Two different lipophilic and amphiphilic bottlebrush block copolymers (BCPs) are placed within shrinking droplets to enable a cooperative working mechanism of microphase segregation and organized spontaneous emulsification, respectively. Layer assemblies of the lipophilic BCP and uniform water nanodroplets stabilized by the bottlebrush surfactant are both generated, and co‐assembled into a bridged lamellar structure with the alternating arrangement of layers and closely packed nanodroplet arrays. Janus microspheres with diverse dual optical characteristics are successfully fabricated, and reflected wavelengths of light are highly tunable simply by changing the formulation or molecular weight of BCP.

Fabrication of a Spontaneous Emulsification System for Low-Permeable Oil Reservoirs and Study of an Enhanced Oil Recovery Mechanism Based on the Nuclear Magnetic Resonance Method

An emulsifier that can spontaneously emulsify oil by a gentle disturbance is an ideal displacement agent for enhanced oil recovery (EOR) in low-permeability reservoirs. Herein, a novel spontaneous emulsification (SE) system with a low concentration of alkali was fabricated in this work. An optimum SE system was screened by interfacial tension (IFT) measurement; the diameter distribution and microscopic morphology of emulsions were observed by an optical microscope and multiple light scattering; the EOR mechanism was studied by nuclear magnetic resonance (NMR) and magnetic resonance imaging (MRI); and the applicability of the SE system in heterogeneous reservoirs was further explored by parallel-core flooding tests. The result showed that the ion pairs formed as a result of the electrostatic attraction of surfactant head groups between cetyltrimethylammonium bromide (CTAB) and nonylphenol polyoxyethylene ether carboxylate (SS-231), which caused the IFT of the CTAB/SS-231 system to reach the ultralow level. The synergistic effect between alkali and surfactants occurred by adding only 0.1 wt % NaOH, which greatly enhanced the capability of emulsification. As a result, the IFT of the SE system (0.4 wt % CTAB/SS-231 + 0.1 wt % NaOH, with massCTAB/massSS-231 = 3:7) could reach 2.1 × 10–3 mN/m, and the SE system could emulsify the crude oil by inverting once. The crude oil in small pores and throats could be displaced by the SE system because the in situ emulsions formed by the SE system were small enough to flow through the small pores; therefore, they exhibited a higher EOR efficiency than the weak emulsification (WE) system. In addition, the SE system showed stronger capabilities in conformance control as a result of the deformation and migration of emulsions. This work revealed the EOR mechanism of the SE system from the level of the pore structure using NMR and MRI, which is important for evaluating the production efficiency of low-permeability reservoirs. Furthermore, the low concentration of alkali (0.1%) in the SE system avoids the scale formation of low-permeability reservoirs effectively.

Diallyl Trisulfide, the Antifungal Component of Garlic Essential Oil and the Bioactivity of Its Nanoemulsions Formed by Spontaneous Emulsification.

The aim of this study was to evaluate the chemical compounds of garlic essential oil (EO), and determine the antifungal efficacy of garlic EO and its major components, diallyl trisulfide and its nanoemulsions against wood-rotting fungi, Trametes hirsuta and Laetiporus sulphureus. GC-MS analysis revealed that the major constituents of garlic EO were diallyl trisulfide (39.79%), diallyl disulfide (32.91%), and diallyl sulfide (7.02%). In antifungal activity, the IC50 value of garlic EO against T. hirsuta and L. sulphureus were 137.3 and 44.6 μg/mL, respectively. Results from the antifungal tests demonstrated that the three major constituents were shown to have good antifungal activity, in which, diallyl trisulfide was the most effective against T. hirsuta and L. sulphureus, with the IC50 values of 56.1 and 31.6 μg/mL, respectively. The diallyl trisulfide nanoemulsions showed high antifungal efficacy against the examined wood-rotting fungi, and as the amount of diallyl trisulfide in the lipid phase increases, the antifungal efficacy of the nanoemulsions increases. These results showed that the nanoemulsions and normal emulsion of diallyl trisulfide have potential to develop into a natural wood preservative.

Stable Encapsulation of Methylene Blue in Polysulfide Organosilica Colloids for Fluorescent Tracking of Nanoparticle Uptake in Cells.

Methylene blue (MB), a century-old drug and a fluorescent dye, has a long history of diverse applications, both in drug therapy and as a tissue-staining agent. However, MB is inherently unstable when exposed to light and reducing agents. In this study, we aim to prepare and characterize polysulfide-based organosilica colloidal particles for efficient, stable, and protective encapsulation of MB. Disulfide- and tetrasulfide-containing organosilane congeners were used as organosilica precursors for direct synthesis of organosilica colloids based on the silica ouzo effect. MB was spontaneously entrapped into the colloidal particles during the particle formation process. The following properties of the colloidal MB were evaluated: particle size, surface charge, atomic distribution, encapsulation efficiency, MB release, photodynamic activity, thiol and ascorbate reactivity, and cytotoxicity. The DLS measurements show that the size of colloidal MB is tunable in a range of 100 nm to 1 μm. SEM images reveal spherical particles with composition-dependent particle sizes of 70–120 nm (coefficient of variation: 15–18%). MB was encapsulated in the colloidal particles with a maximal efficiency of 95%. The release of MB from the colloids was <1% at 4 h and <3.5% at 48 h. The colloidal particles show much reduced photodynamic activity, low reactivity toward reducing agents, and low cytotoxicity. Accordingly, the colloidal MB was proposed and further investigated as a fluorescent particle tracer for the study of cell–nanoparticle interactions. In conclusion, MB can be efficiently and stably loaded into polysulfide organosilica colloidal particles using a simple and convenient physical route.

Artificial Inclusion Environments—Replicating Industry in the Laboratory

The authors present a series of complementary test methods which were developed and used to investigate reactions between high aluminium steel and silica rich inclusions. Non-metallic inclusions (NMIs) cause many defects in the final steel product, therefore the ability to track their size, morphology and composition and correlate this with fundamental reaction kinetics provides important knowledge to support the production of clean quality steel products. Novel steel grades such as TRIP, TWIP and low-density steels have high aluminium contents; aluminium is a readily oxidisable species presenting the potential for instability and excessive reaction with commonly used mould powders that contain silica. A novel combination of techniques including HT-CLSM (High-Temperature Confocal Laser Scanning Microscope), XCT (X-ray computed tomography) and SEM/EDS (scanning electron microscopy/electron dispersive spectroscopy) have been used to study the interaction of entrained mould powder inclusions with steel at high temperatures simulating industrial conditions. This report presents a discussion on the development of techniques and samples to achieve representative and repeatable results that can provide information on the complex chemical and physical interaction phenomena with confidence. Each experimental technique had its own learning points and consequent results. Outcomes presented include possible confirmation of the chemical reaction rate controlling step being aluminium mass transfer; heterogeneous local environmental conditions including fluidity and chemical composition; and occurrence of spontaneous emulsification where the mould powder inclusion breaks apart into a cloud of smaller fragments.