Controllable preparation and formation mechanism of BSA microparticles using supercritical assisted atomization with an enhanced mixer

Supercritical fluid assisted atomization introduced by a hydrodynamic cavitation mixer (SAA-HCM) was used to prepare bovine serum albumin (BSA) microparticles. Water was used as the sole solvent. A hydrodynamic cavitation mixer was applied to improve mass transfer and achieve a continuous near-thermodynamic-equilibrium solubilization of SC-CO₂ in the liquid solution. Under the different conditions, the prepared BSA microparticles had various morphologies, such as corrugated particles, smooth hollow spherical particles and cup particles, with particle diameters ranging from 0.3 to 5 μm. The microparticle formation process was elucidated with the shell formation and central bubble mechanism. Compared to native BSA, BSA microparticles did not show significant change in primary structure, according to the results of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The secondary structure of BSA was characterized by Fourier transform infrared spectroscopy (FT-IR). No new peaks were observed after SAA-HCM processing. In addition, the crystalline structure of the BSA microparticles was demonstrated to be amorphous because of the sudden supersaturation in the precipitation process. The SAA-HCM process is expected to be a promising technique for producing microparticles suitable for pulmonary delivery of therapeutic macromolecules.     

Influence of pressure, temperature and concentration on the mechanisms of particle precipitation in supercritical antisolvent micronization

In this work, supercritical antisolvent micronization (SAS) is used to produce nanoparticles, microparticles and expanded microparticles of a model compound, gadolinium acetate (GdAc), using dimethylsulfoxide (DMSO) as the liquid solvent with the aim of studying the dependence of particles’ diameter and morphology on some process parameters like pressure, temperature and concentration of the starting solution. Experiments are performed varying the precipitation pressure between 90 and 200 bar, the precipitation temperature between 35 and 60 °C and the concentration of GdAc in the liquid solution in the range from 20 to 300 mg/mL. The experimental evidences show that the formation of particles with specific sizes in the micrometric and nanometric range depends on specific values of each one of these parameters. An explanation of the results is proposed in terms of the competition between two characteristic times of the SAS process that can control the precipitation process. The time of jet break-up of the liquid solution that produces liquid droplet formation, and the dynamic surface tension vanishing time, that induces gas mixing with the precipitation of nanoparticles from the gaseous phase. Indeed, GdAc sub-microparticle, or microparticle (diameter in the range 0.23-1.6 μm with mean diameters in the range 0.28-0.52 μm) formation can be attributed to micro-droplet drying, whereas nanoparticles (mean diameter in the range 90-210 nm) are consistently produced when gas mixing is the possible governing process. In conclusion, the precipitation mechanisms can be modulated varying one SAS parameter a time, thus selecting the range of particle diameters required for the specific application.     

Biodiesel production using supercritical methanol/carbon dioxide mixtures in a continuous reactor

Fatty acid methyl esters (biodiesel) were produced by the transesterification of triglycerides with compressed methanol (critical point at 240 °C and 81 bar) in the presence of solid acids as heterogeneous catalyst (SAC-13). Addition of a co-solvent, supercritical carbon dioxide (critical point at 31 °C and 73 bar), increased the rate of the supercritical alcohols transesterification, making it possible to obtain high biodiesel yields at mild temperature conditions. Experiments were carried out in a fixed bed reactor, and reactions were studied at 150-205 °C, mass flow rate 6-24 ml/min at a pressure of 250 bar. The molar ratio of methanol to oil, and catalyst amount were kept constant (9 g). The reaction temperature and space time were investigated to determine the best way for producing biodiesel. The results obtained show that the observed reaction rate is 20 time faster than conventional biodiesel production processes. The temperature of 200 °C with a reaction time of 2 min were found to be optimal for the maximum (88%) conversion to methyl ester and the free glycerol content was found below the specification limits.     

Use of solvent mixtures in supercritical antisolvent process to modify precipitates morphology: Cellulose acetate microparticles

In the supercritical antisolvent precipitation (SAS), the jet fluid dynamics is characterized by two-phase mixing at subcritical conditions, and by one-phase mixing at completely developed supercritical conditions. The amplitude of the pressure range, in which binary systems organic solvent/scCO₂ exhibit the transition between two-phase to one-phase mixing, depends on the organic solvent that is in contact with supercritical carbon dioxide (scCO₂) and conditions the morphology of the microparticles produced by SAS. When this pressure range is wide, as in the case of dimethylsulfoxide (DMSO), solutes solubilized in the organic solvent can be precipitated as microparticles by atomization, droplets formation and drying; when this pressure range is narrow, as for acetone, gas mixing prevails and only nanoparticles are generally observed. Therefore, generally speaking, solutes that are soluble only in solvents exhibiting gas mixing in scCO₂, do not exhibit microparticles morphology and this fact is a limitation for several industrial applications.In this work, a model compound, cellulose acetate (CA), that is slightly soluble in DMSO and freely soluble in acetone, was processed by SAS using mixtures of the two solvents that exhibit intermediate behaviors between the two pure solvents, to extend two phase mixing and produce CA microparticles. Using different DMSO/acetone mixture percentages, the effects of the polymer concentration in the liquid solution and of the pressure were studied. A mixture of DMSO/Acetone 50/50 (v/v), at a pressure of 85 bar and a concentration of the liquid solution equal to 40 mg/mL, efficiently produced non-coalescing CA microparticles with a mean diameter of 0.42 μm and a standard deviation of about 0.15 μm, demonstrating that this SAS strategy can be successful.     

Particle formation of an edible fat (rapeseed 70) using the supercritical melt micronization (ScMM) process

The supercritical melt micronization (ScMM) process, also known as particles from gas saturated solutions (PGSS) was applied, in a continuous operated pilot plant, for the particle formation of the edible fat, rapeseed 70 (RP70). The effect of variables like the CO₂ concentration, the melt temperature and the atomization pressure were studied in order to investigate particle morphology, density and the particle size distribution. The experiments were performed at CO₂ concentrations between 0 and 50 wt%, atomization pressure between 70 and 180 bar and melt temperature between 60 and 100 °C. Particles obtained as a function of the CO₂ concentration, showed completely solid, spherical–hollow and aggregated particles with a decrease in particle mean size as the concentration of CO₂ was increased. The results obtained as a function of atomization pressure showed no significant influence on particle morphology and size distribution. Experiments carried out as a function of the melt temperature showed distorted, spherical–hollow and aggregated particles. Furthermore, a theory was developed to explain the mechanism for particle formation as a function of the CO₂ concentration and the melt temperature. The crystallinity of the final product of RP70, showed an alpha polymorph with a crystallinity of 84%.     

Kinetic Modeling of the Gas Antisolvent Process for Synthesis of 5‐Fluorouracil Nanoparticles

Mathematical modeling for 5‐fluorouracil (5‐FU) nanoparticle synthesis via gas antisolvent (GAS) process was investigated. 5‐FU was precipitated from a dimethyl sulfoxide (DMSO) solution using CO₂ as antisolvent. The particle size was controlled by nucleation and growth rates, therefore, the kinetic modeling study is essential. Thermodynamic modeling was applied to determine optimal operating conditions for experimental 5‐FU synthesis. Kinetic parameters were evaluated by fitting the particle size distribution predicted by the model to experimental data. The experimental and modeling results indicated that the particle size decreased with increasing the antisolvent addition rate.     

High-pressure phase equilibria of systems carbon dioxide + n-eicosane and propane + n-eicosane

In this work we investigated the phase equilibrium behavior of the binary asymmetric systems propane (C3) + n-eicosane (C20) and carbon dioxide (CO₂) + n-eicosane (C20). We used a variable-volume view cell for obtaining fluid–fluid equilibrium (FFE), solid–fluid equilibrium (SFE) and solid–fluid–fluid equilibrium (SFFE) experimental data. We modeled the phase equilibria of both systems using the Peng–Robinson Equation of State for describing the fluid phases and an expression for the fugacity of pure solid n-eicosane with parameters fit to reproduce the pure n-eicosane melting line. We performed the phase equilibrium calculations by implementing path-following methods for tracking entire solid–fluid (SF) and solid–fluid–fluid (SFF) equilibrium curves for binary asymmetric mixtures. This made it possible to obtain complete isoplethic lines or complete three-phase equilibrium lines in single runs. Although the model is relatively simple, it is able to grasp the complex observed behavior for the systems studied here.     

Supercritical extraction of Hypericum caprifoliatum using carbon dioxide and ethanol + water as co-solvent

Hypericum caprifoliatum is a natural source of phloroglucinol derivatives such as hyperbrasilol B, uliginosin B and japonicin A, compounds with pharmaceutical interest due to principally their antidepressant properties. This work studies supercritical fluid extraction to obtain non-volatile compounds from the aerial parts of H. caprifoliatum as well as the influence of the co-solvent as agent of process intensification. The experimental procedures were carried out in a pilot unity. The mathematical simulation of the experimental data was performed by mean of three mathematical models aiming to promote the improvement of the knowledge about this technology. The feasibility of the mathematical model was investigated by mean of fitting of the experimental data obtained. The co-solvents tested were ethanol, water, and water–ethanol mixtures.     

Non-invasive quantification of phase equilibria of ternary mixtures composed of carbon dioxide,organic solvent and water

Raman spectra of ternary mixtures of unknown composition are processed to extract the content of the compounds in the mixture. The evaluation method of the ternary mixtures is based on a previous calibration of binary mixtures. The applicability of the method is demonstrated by measuring the vapour liquid equilibria of the ternary system carbon dioxide, ethyl acetate and water at 8.5 MPa and 310 K and providing them in a triangle diagram. The advantages of the used flexible Raman sensor in comparison to conventional techniques for the determination of phase equilibria are discussed.     

Extraction of jojoba oil with liquid CO2 + propane solvent mixtures

In this work liquid CO₂/propane mixtures were used to extract jojoba oil from oilseeds. First, experiments at 313 K and pressures of 70 bar and 200 bars were carried out on jojoba oil deposited on glass spheres, using different solvent concentrations (30 wt%, 50 wt% and 70 wt% CO₂), to assess the influence of the solvent composition and phase behavior on the extraction rate. Then, jojoba oil was extracted from the milled seeds under homogeneous liquid conditions, using solvent mixtures containing 30 wt% and 50 wt% CO₂ at 70 bar and 313 K. A solvent mixture with 30 wt% CO₂ exhibited good solvent power. Oil extraction yields of 98% were obtained using a minimum solvent to oilseed mass ratio of 5 g solvent/g oilseed and operating the extractor at 313 K and 70 bar.     

Deposition of Pd into mesoporous silica SBA-15 using supercritical carbon dioxide

Pd was deposited into mesoporous silica SBA-15 using supercritical CO₂ (scCO₂). Palladium hexafluoroacetylacetonate [Pd(hfac)₂] was dissolved in scCO₂ and impregnated into the support at very mild conditions, 40 °C and 85 bar. Then the organometallic precursor was reduced with H₂ in the CO₂ mixture or, after depressurization, in pure H₂. Materials were characterized by TGA, XRD, TEM, SEM, EDX, ICP-OES and N₂-adsorption experiments. Pd nanoparticles evenly distributed into the support (1-3 mol% Pd by ICP-OES) are only obtained when the reduction is performed in pure H₂. Cluster size is limited in two dimensions by the pore size of the support but clusters grow larger with increasing impregnation time and turn into small nanowires. The catalytic activity of the Pd/SiO₂ composite material was confirmed following the reduction of 4-nitrophenol to 4-aminophenol in water by UV-vis spectroscopy.     

On-line H NMR spectroscopic investigation of hydrogen bonding in supercritical and near critical CO2–methanol up to 35 MPa and 403 K

On-line NMR spectroscopy can beneficially be applied to studies of supercritical and near-critical fluids as an alternative to optical spectroscopy. Up to now high pressure NMR experiments are predominantly accomplished using custom made NMR batch reactors. The authors present a novel high pressure cell with displacement plunger for on-line NMR experiments on compressible fluids which can be used in conjunction with commercially available SCF NMR flow probes. The on-line technique offers advantages compared to stopped flow techniques such as enhanced control of mixture composition and reaction parameters as well as the facility of engagement into the reaction. The new apparatus is used for NMR studies on hydrogen bonding of methanol in near critical and supercritical carbon dioxide up to 403 K and 35 MPa for which data on the chemical shift of the hydroxyl group and methyl group are reported and interpreted.     

Impregnation of vitamin E acetate on silica mesoporous phases using supercritical carbon dioxide

Impregnation of a drug model (α-tocopheryl acetate) into mesoporous host matrices has been carried out using supercritical carbon dioxide (SC CO₂) as impregnation solvent at 15 MPa and 313 K with a flow rate of 500 g h⁻¹. The operating conditions were defined following the solute concentration in the fluid phase as a function of pressure and carbon dioxide flow rate. Solubility measurements of α-tocopheryl acetate were first performed at 313 K for pressures ranging 10-20 MPa. High values of solubility in SC CO₂ were measured: 6 wt% at 10 MPa and 14 wt% at 20 MPa. Measurements of the concentration of the solute in SC CO₂ in the experimental conditions of impregnation in dynamic mode showed than it was ten times lower than the solubility. The variations of this concentration have been studied at 313 K, for a pressure varying from 8 to 15 MPa, and for a carbon dioxide flow rate varying from 120 to 600 g h⁻¹. Two different host matrices were used: a commercial chromatographic silica support and a MCM-41-type mesoporous organized silica synthetized at the laboratory. This latter showed the best drug loading of 1.14 g per gram of adsorbent. The drug loadings obtained in supercritical media were similar to the ones obtained in liquid media using hexane as impregnation solvent. Nevertheless, the maximum loading was obtained after 1 h of impregnation in SC media while 4 h were needed in liquid media.     

Manipulating the size,the morphology and the polymorphism of acetaminophen using supercritical antisolvent (SAS) precipitation

The supercritical antisolvent technology is used to crystallize paracetamol particles. Supercritical carbon dioxide (scCO₂) is used as antisolvent. Ethanol, acetone and mixtures of ethanol and acetone are used as solvents. The initial concentration of paracetamol in the solution was varied between 1 and 5 wt%, the composition of the ethanol/acetone solvent mixture between 50 and 90 wt% of ethanol and the operation pressure between 10 and 16 MPa at a temperature of 313 K. The most important finding is that the polymorph of paracetamol crystals can be adjusted between monoclinic and orthorhombic by varying the content of ethanol in the solution. The second important finding is that the occurrence of primary and secondary crystal structures can be explained solely by the overall supersaturation during the crystallization process. While X-ray diffraction was used to analyze the polymorph of the particles, their morphology was analyzed using scanning electron microscopy.     

Foaming strategies for bioabsorbable polymers in supercritical fluid mixtures. Part I. Miscibility and foaming of poly(l-lactic acid) in carbon dioxide + acetone binary fluid mixtures

Miscibility and foaming of poly(l-lactic acid) (PLLA) in carbon dioxide + acetone mixtures have been explored over the temperature and pressure ranges from 60 to 180 °C and 14 to 61 MPa. Liquid-liquid phase boundaries were determined in a variable-volume view-cell for polymer concentrations up to 25 wt% PLLA and fluid mixtures containing 67-93 wt% CO₂ over a temperature range from 60 to 180 °C. Even though not soluble in carbon dioxide at pressures tested, the polymer could be completely solubilized in mixtures of carbon dioxide and acetone at modest pressures.Foaming experiments were carried out in different modes. Free-expansions were carried out by exposure and swelling in pure carbon dioxide in a view-cell followed by depressurization. Foaming experiments were also carried out within the confinement of specially designed molds with porous metal surfaces as boundaries to direct the fluid escape path and to generate foams with controlled overall shape and dimensions. These experiments were conducted in pure carbon dioxide and also in carbon dioxide + acetone fluid mixtures over a wide range of temperatures and pressures. Foaming in carbon dioxide + acetone mixtures was limited to 1 and 4 wt% acetone cases. Microstructures were examined using an environmental scanning electron microscope (ESEM). Depending upon the conditions employed, pore diameters ranging from 5 to 400 μm were generated. At a given temperature, smaller pores were promoted when foaming was carried out by depressurization from higher pressures. At a given pressure, smaller pores were generated from expansions at lower temperatures. Foams with larger pores were produced in mixtures of carbon dioxide with acetone.     

Preparation of poly(L-lactic acid) submicron particles in aerosol solvent extraction system using supercritical carbon dioxide

The aerosol solvent extraction system process (ASES), which is one of the supercritical anti solvent processes (SAS), was used to produce poly(L-lactic acid) (PLLA) into the submicron particles. Dichloromethane (DCM, CH₂Cl₂) and carbon dioxide were selected as a solvent and as an antisolvent for PLLA, respectively. The objective of this study was to investigate the effect of the various process parameters such as temperature, pressure, and solution concentration on PLLA particles. With increasing temperature and pressure, particle size was increased. Also, higher PLLA concentration led to larger particle size and broader particle size distribution. A scanning electron microscope (SEM) was used to observe the morphology and size of PLLA particles recrystallized by ASES process. The mean particle size and its distribution of processed particles were measured by using a laser diffraction particle size analyzer (PSA).     

Phase behavior of the poly(neopentyl methacrylate) + supercritical fluid solvents + neopentyl methacrylate system and CO2 + neopentyl methacrylate mixtures at high pressure

High-pressure phase behaviors are measured for the CO₂ + neopentyl methacrylate (NPMA) system at 40, 60, 80, 100, and 120 °C and pressure up to 160 bar. This system exhibits type-I phase behavior with a continuous mixture-critical curve. The experimental results for the CO₂ + NPMA system are modeled using the Peng-Robinson equation of state. Experimental cloud-point data up to the temperature of 180 °C and the pressure of 2000 bar are presented for ternary mixtures of poly(neopentyl methacrylate) [poly(NPMA)] + supercritical solvents + NPMA systems. Cloud-point pressures of poly(NPMA) + CO₂ + NPMA system are measured in the temperature range of 60-180 °C and to pressures as high as 2000 bar with NPMA concentration of 0.0, 5.2, 19.0, 28.1 and 40.2 wt%. It appears that adding 51.2 wt% NPMA to the poly(NPMA) + CO₂ mixture does significantly change the phase behavior. Cloud-point curves are obtained for the binary mixtures of poly(NPMA) in supercritical propane, propylene, butane, 1-butene, and dimethyl ether (DME). The impact of dimethyl ether concentration on the phase behavior of the poly(NPMA) + CO₂ + x wt% DME system is also measured at temperature of 180 °C and pressure range of 36-2000 bar. This system changes the pressure-temperature (P-T) slope of the phase behavior curves from upper critical solution temperature (UCST) region to lower critical solution temperature (LCST) region as the NPMA concentration increases.     

Extraction and identification of proanthocyanidins from grape seed (Vitis Vinifera) using supercritical carbon dioxide

In this study, supercritical carbon dioxide extraction of proantocyanidins (PRCs) was performed and the effect of different pressure, temperature and ethanol percentage was investigated. High performance liquid chromatography was used for the analysis of the compounds and it was found that the most effective parameter on the extraction was the amount of the ethanol percentage. Each compound was extracted from grape seeds at their maximum level when different parameters were used which was probably because of their different polarities. Gallic acid (GA), epigallocatechin (EGC) and epigallocatechingallate (EGCG) were extracted at their maximum level when the 300 bar 50 °C and 20% of ethanol was used. The maximum amount of catechin (CT) and epicatechin (ECT) were obtained when 300 bar 30 °C and 20% of ethanol was used for extraction, and 250 bar, 30 °C and 15% of ethanol was needed to extract highest amount of epicatechingallate (ECG).     

Pervaporation separation of isopropanol/benzene mixtures using inorganic–organic hybrid membranes

A series of pervaporation hybrid membranes were prepared from polyethylene glycol (PEG) and phenylaminomethyl trimethoxysilane (PAMTMS) based on the sol‐gel process, in which PEG was used as an organic moiety to improve the affinity for organic alcohols and silicone of PAMTMS was used as inorganic moiety to increase the permeation flux of organic species. Their application to separate isopropanol/benzene mixtures was investigated. FTIR spectra confirmed the reaction products. DSC measurement revealed that the influence of PEG content on the T_g and thermal behavior of membranes A, B, and C. FE‐SEM images exhibited that phase‐separated structure has occurred when the PEG content elevated to some extent. Pervaporation experiments showed that the permeation flux increased and the separation factor decreased with an increase in isopropanol (IPA) content in feed at 30°C. Meanwhile, the separation factor increased with an increase in feed temperature at 60 vol % IPA content. Moreover, it was found that the permeation flux was independent of the feed temperature, suggesting that feed temperature has little impact on the thermal motion of polymer chains. The increasing cross‐linking degree in hybrid matrix might be responsible for such trend. Based on these findings, it can be concluded that these pervaporation hybrid membranes have potential applications in the separation of isopropanol/benzene binary mixtures. © 2010 Wiley Periodicals, Inc. J Appl Polym Sci, 2010     

Debinding behaviors of injection molded ceramic bodies with nano-sized pore channels during extraction using supercritical carbon dioxide and n-heptane solvent

Debinding behaviors related on changes in capillary pore structure during extraction with supercritical carbon dioxide and n-heptane, respectively were investigated for injection molded ceramic bodies consisting of skeleton pores of 68 nm. For the debinding processes, both debinding curves showed a square root of time dependence but significantly deviated in the middle or end period of debinding because of structural changes with pores during extraction. The debinding bodies experienced capillary changes having a debinding front separating the undebinded region with fluid state into the debinding region with pendular state in the wax-based green bodies. The debinding rate of the supercritical fluid extraction was five times higher than that of the solvent extraction because of a higher interdiffusion diffusivity and a formation of relatively large pore channels. An abrupt change of pore structures between debinded and undebinded region in the green bodies caused severe defects during the solvent extraction even at a low rate of debinding at 313.15 K, while the severity of the capillary changes was overcome during the supercritical fluid extraction and the debinded ceramic bodies were free from defects even at a high rate of debinding at 328.15 K. It is attributed to a reduction of the capillary stress developed on debinding front during the supercritical fluid extraction.