In-line Mixer Research Articles

Radical Protein Footprinting in Mammalian Whole Blood.

Hydroxyl Radical Protein Footprinting (HRPF) is a powerful method to probe the solvent-accessible surface area of proteins. It is mostly used to study the higher-order structure of proteins, as well as protein-protein and protein-carbohydrate interactions. Hydroxyl radicals are generated by the photolysis of hydrogen peroxide and these radicals modify the surface amino acids. Bottom-up proteomics is then applied and peptide oxidation is calculated and correlated with solvent accessibility. It is mainly performed in vitro ; however, it has been recently used in living systems, including live cells, live nematodes, and 3D cell cultures. Mammalian tissues are still out of reach as they absorb UV strongly, hindering radical generation. Here, we describe the first example of RPF in mammalian stabilized whole blood. Using photoactivation of persulfate with a commercially available FOX Photolysis System modified for sample handling and inline mixing, we demonstrate the first labeling of proteins in whole blood. We demonstrate that the RPF protocol does not alter the blood cell gross morphology outside of a moderate hypertonicity equivalent to sodium chloride exposure prior to labeling. We detail an improved quenching protocol to limit background labeling in persulfate RPF. We describe the labeling of the top ten most abundant proteins in the blood. We demonstrate the equivalence of ex vivo labeling in whole blood with labeling of the same structure in vitro using hemoglobin as a test system. Overall, these results now open the possibility of performing RPF-based structural proteomics in pre-clinical models and using readily available clinical samples.

Novel Fast Cure Silicone Inks for Single‐Step, Support‐Free 3D Printing of Tall, Overhanging, and High Aspect Ratio Structures

AbstractSilicone elastomers have a broad variety of applications, such as soft robotics, biomedical devices, and structural metamaterials. The extrusion‐based method known as direct ink write (DIW) has enabled the production of additively manufactured silicone structures. However, this method is limited to manufacturing mostly planar or pseudo‐3D structures. Due to the low self‐supporting capabilities of extruded strands for traditional silicone‐based “inks,” obtaining tall or overhanging structures, or structures comprised by thin walls is not feasible. Here, a novel Fast Cure silicone‐based ink is demonstrated that enables manufacturing of complex 3D structures. The Fast Cure ink is a two‐part mixture and silicone structures are produced by inline mixing and coextrusion of a part containing a catalyst (part A) and a part containing a crosslinker (part B). By the virtue of crosslinking, the extruded strands rapidly rigidize, increasing their self‐supportive capacity. Hence, structures can be obtained with superior shape retention and previously unobtainable parts are realized that are tall, with aspect ratios higher than 3, and have overhanging features, achieving inclination angles smaller than 35° with respect to the printing plane. These minimal sag parts are achieved without requiring extra curing or mechanisms, support structures, or suspension baths.

Modelling W/O/W double emulsions preparation in static mixers: Accounting for the shear-thinning behaviour of the dispersed phase

The dispersion of a shear-thinning non-Newtonian fluid using in-line mixers to prepare double emulsions is investigated experimentally and numerically in this study. A two-step method is adopted for the preparation of double emulsions, comprising in a first step the preparation of a reverse water-in-oil (W/O) emulsion using a high-shear rotor-stator device. This inner emulsion is dispersed in a second step into an outer aqueous phase by continuous parallel pumping of both phases through a set of static mixers. The second preparation step is modelled using a population balance equation (PBE) accounting for droplet breakage inside the mixers. Since the inner emulsion behaves as a shear-thinning fluid, the breakage efficiency depends on the local shear rate. Using classical turbulent breakage kernels is therefore inappropriate, hence there is a need for a kernel accounting for the shear-thinning character of the dispersed phase. To tackle this issue, single-phase CFD simulations of the fluid flow through the mixers are carried out showing a high shear rate magnitude near the walls of the pipe and the mixer crossbars. To avoid CFD-PBE direct coupling, the probability density function (PDF) of the shear rate is extracted from the CFD simulations. This PDF is then used to develop a volume-averaged PBE, where a classical Ostwald-de Waele model is employed to relate the apparent viscosity of the inner emulsion to the shear rate. Different experimental parameters influencing the quality of the dispersion are investigated, including the fraction of the inner emulsion phase, the oil viscosity and the inner and outer aqueous phase viscosities. In the different cases, the model captures the transient evolution of the droplet size distribution of the W/O/W double emulsions at the outlet of the mixers while the computational time remains very low.

Streamlining regular liquid chromatography with MALDI-TOF MS and NMR spectroscopy using automatic full-contact splitless spotting interface and flash-tap fractioning collection

BackgroundHigh-resolution matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) and nuclear magnetic resonance (NMR) spectroscopy are powerful tools to identify unknown psychoactive substances. However, in complex matrices, trace levels of unknown substances usually require additional fractionation and concentration. Specialized liquid chromatography systems are necessary for both techniques. The small flow rate of nano LC, typically paired with MALDI-TOF MS, often results in prolonged fractionation times. Conversely, the larger flow rate of semi-preparative LC, used for NMR analysis, can be time-consuming and labor-intensive when concentrating samples. To address these issues, we developed an integrated automatic system that integrated to regular LC. ResultAutomatic spot collector (ASC) and automatic fraction collector (AFC) were present in this study. The ASC utilized in-line matrix mixing, full-contact spotting and real time heating (50 °C), achieving great capacity of 5 μL droplet on MALDI plate, high recovery (76–116%) and rapid evaporation in 2 min. The analytes were concentrated 4–8 times, forming even crystallization, reaching the detection limit at the concentration of 50 μg L−1 for 12 psychoactive substances in urine. The AFC utilizes flexible tubing which flash-tapped the microtube's upper rim (3 mm depth) instead of reaching the bottom. This method prevents sample loss and minimizes the robotic arm's movement, providing a high fractionating speed at 6 s 12 psychoactive compounds were fractionated in a single round analysis (recovery: 81%–114%). Methamphetamine and nitrazepam obtained from drug-laced coffee samples were successful analyzed with photodiode array (PDA) after one AFC round and NMR after five rounds. SignificanceThe ASC device employed real-time heating, in-line matrix mixing, and full-contact spotting to facilitate the samples spotting onto the MALDI target plate, thereby enhancing detection sensitivity in low-concentration and complex samples. The AFC device utilized the novel flash-tapping method to achieve rapid fractionation and high recovery rate. These devices were assembled using commercially available components, making them affordable (400 USD) for most laboratories while still meeting the required performance for advanced commercialized systems.

Continuous production of highly bioavailable lycopene nanodispersions via subcritical ethanol extraction and in-line mixing

This study aims to prepare Z-isomer-rich lycopene nanodispersions via ethanol extraction at higher temperature and pressure conditions (i.e., subcritical conditions) from tomato powder and continuous dispersion using an in-line swirl mixer. Firstly, the effects of ethanol extraction conditions, i.e., temperature (40–160 ℃), time (10–60 min), and pressure (10–30 MPa), on lycopene recovery and Z-isomerization were investigated. The extraction at > 120 ℃ significantly enhanced lycopene recovery and Z-isomer ratio of lycopene in the resulting extract. Subsequently, the resulting extract liquid (ethanol solution) was continuously dispersed in water containing polyglycerol fatty acid ester using an in-line mixer, and successfully prepared Z-isomer-rich (approximately 70%) lycopene nanodispersions. This production process is economical and highly compatible with the food industry due to using a safe extraction solvent and continuous production capability. In the future, it will be necessary to investigate the biological activities of the resulting lycopene and perform scale-up verification of this process.

Enhancing the Techno-Functional Properties of Lentil Protein Isolate Dispersions Using In-Line High-Shear Rotor-Stator Mixing.

In response to global challenges such as climate change and food insecurity, plant proteins have gained interest. Among these, lentils have emerged as a promising source of proteins due to their good nutritional profile and sustainability considerations. However, their widespread use in food products has been impeded by limited solubility. This study aimed to investigate the potential of high-shear mixing, a resource-efficient technique, to enhance lentil protein solubility and its functional properties. Red lentil protein isolate powders were rehydrated and subjected to a semi-continuous in-line high-shear treatment at 10,200 rpm for a timespan ranging from 0 to 15 min. The results highlighted a significant (p < 0.05) increase in solubility from 46.87 to 68.42% after 15 min of shearing and a reduction in particle size as a result of the intense shearing and disruption provided by the rotor and forced passage through the perforations of the stator. The volume-weighted mean diameter decreased from 5.13 to 1.72 µm after 15 min of shearing, also highlighted by the confocal micrographs which confirmed the breakdown of larger particles into smaller and more uniform particles. Rheological analysis indicated consistent Newtonian behaviour across all dispersions, with apparent viscosities ranging from 1.69 to 1.78 mPa.s. Surface hydrophobicity increased significantly (p < 0.05), from 830 to 1245, indicating exposure of otherwise buried hydrophobic groups. Furthermore, colloidal stability of the dispersion was improved, with separation rates decreasing from 71.23 to 24.16%·h-1. The significant enhancements in solubility, particle size reduction, and colloidal stability, highlight the potential of in-line high-shear mixing in improving the functional properties of lentil protein isolates for formulating sustainable food products with enhanced techno-functional properties.

An experimental study of localized flow pattern, strain rate and turbulence kinetic energy dissipation rate of in-line teethed high shear mixer with different rotor and stator structures

The in-line teethed high shear mixers (HSMs) have been widely used in the chemical industry. However, there is still a lack of comprehensive understanding of the hydrodynamics characteristics about the attractive process equipment. In this paper, the effects of rotor and stator structures and rotor speed on flow characteristics of in-line teethed HSM are systematically studied by the particle image velocimetry experiment for the first time. The results show that increasing the rotor speed can significantly increase the strain rate (Sxy) and turbulence kinetic energy dissipation rate (ε). Especially, when the shear gap width is reduced from 1.5 mm to 0.5 mm, the mean ε increases by 96.8 %. Furthermore, two dimensionless correlations are established between the maximum value of the Sxy, ε and rotor and stator geometrical parameters to provide a valuable guidance for the geometrical design and optimization of in-line teethed HSM.

Energy Conversion Characteristics of an In-Line Mixing Pump in Solid–Liquid Two-Phase Dynamic Mixing

Energy Conversion Characteristics of an In-Line Mixing Pump in Solid–Liquid Two-Phase Dynamic Mixing

CFD simulation of an axial discharge rotor-stator mixer: LES versus RANS predictions

The large eddy simulation (LES) technique is used to model the turbulent flow of water in a two-stage, axial discharge, in-line rotor-stator mixer. The WALE sub-grid scale model is employed, and the resulting filtered velocity field is ensemble averaged to predict the mean velocity, turbulent kinetic energy, energy dissipation rate, and energy production rate. The results of these 3-D sliding mesh simulations are compared to analogous RANS predictions on similar meshes using the realizable k-ε turbulence model. Stator slot flow rate and rotor torque/power draw are also monitored. RANS and LES predict similar mean velocity profiles, flow rate and rotor torque. LES predicts higher turbulent kinetic energy, but lower dissipation rate compared to RANS. Results from a coarse and refined computational mesh are compared to discuss the significance of resolving smaller scales for LES. The effect of operating conditions on throughput, power draw, and other quantities of practical utility are also discussed. It was found that prediction of macroscopic quantities such as flow rate and power draw can be done efficiently using either RANS or LES on a relatively course mesh while capturing turbulence quantities requires a well refined LES simulation.

Numerical Investigation of Hydrodynamics in Inline Mixers

The utilization of inline mixers for continuous fluid mixing is a common practice in chemical industries. Information related to hydrodynamics condition in inline mixers is found to be limited while it is crucial for inline mixer type selection. Evaluation of hydrodynamic characteristics in several commercial inline mixers is analyzed in this study, including Y-type, T-type, elbow type, and internal injection mixer types. The analysis is conducted using computational fluid dynamics (CFD) for various Reynolds number conditions from 5,000 to 200,000. Mesh independence tests and model validation with experimental results are performed to ensure simulation accuracy. The CFD results showed that the Y-type mixer type provides the shortest distance i.e., about 12 times diameter. The internal injection type mixer has the most extended hydrodynamic region length to reach a fully-developed region for the same Reynolds number. The highest wall stress is indicated by the injection inline mixer type that located at the outer side wall of the injection pipe. The wall stress at that region is observed to be around twice as large compared to the other mixer type. Meanwhile, the mixer joint in the T-type and Y-type mixers were found to have a wall stress hotspot. Logarithmic correlation formulas were successfully formulated to correlate hydrodynamic region length and Reynolds number for all mixer types. The study results are essential in serving as the basis for inline mixer selection in the industrial sector

Set-on-demand of alkali-activated slag mixture using twin-pipe pumping

To achieve the set-on-demand purpose of concrete for digital fabrication where the opposing requirements for pumpability and buildability exist during the process, an inline mixing strategy, named twin-pipe pumping, has been developed utilizing a static mixer to blend two separate streams to accelerate setting right before extrusion. The system was initially developed utilizing two streams including a Portland cement-based mixture (without accelerator) and a limestone powder-based mixture (with an accelerator but without cement). However, the high content of cement may result in higher material costs along with lower sustainability, given the fact that the production of Portland cement contributes to around 8% of global CO2 emissions. This study aims to develop an alkali-activated slag mixture to increase the sustainability in digital fabrication, using twin-pipe pumping. The constituent mixtures consisting of a slag-based (without alkali) and a limestone powder-based (with varying alkali content) mix were formulated and their rheological behavior was measured for good pumpability. Phase transition from liquid to solid phase, mechanical strength, and pore structure was evaluated. Twin-pipe pumping was used in a printing trial. Results showed that the alkali-activated slag mixture achieved fast phase transition for set-on-demand purposes by combining the alkali in the limestone-based mixture with the slag in the slag-based mixture. Increasing the NaOH concentration or incorporating nano clay improved the structural build-up for high buildability.

Stream Water Quality Control and Odor Reduction through a Multistage Vortex Aerator: A Novel In Situ Remediation Technology

In this work, we report the restoration of a polluted urban stream by employing the multistage vortex aerator (MVA), an in-line mixer device that improves the dissolved oxygen concentration of polluted streams and accelerates the water purification rate. It was observed during the field experiment that the dissolved oxygen was enhanced up to 7.05 mg/L and the water quality was improved to a good grade. As a result, the complex odor was successfully eliminated and reduced by up to 71.9%, while the water quality grade was also improved by more than two grades on average. Stream water quality indicators monitored for twelve months revealed high removal rates of total phosphorous (56.4%) and suspended solids (61%). The study demonstrated MVA as a promising eco-friendly technology for significant improvement in urban stream water quality. Moreover, the MVA process creates no secondary pollution and is believed to be a sustainable treatment option for odorous water bodies. Overall, the MVA process is technically feasible for implementation, and this study provides a specific reference as a basis for the treatment of polluted water bodies in urban settings.

The Use of A Static Mixer for The Coagulation Unit in The Duren Seribu II Water Treatment Plant

In the drinking water treatment plant involves a significant component i.e., coagulation, which distributed coagulants vastly and equally through rapid stirring for destabilizing colloids and suspended particles in the raw water. In water treatment plants, coagulation units are often classified into mechanical and hydraulic coagulation. This study aimed to discover the use of and in-line static mixers as coagulation in designing the Duren Seribu II Drinking Water Treatment Plant (WTP). The design criteria for coagulation unit in Duren Seribu II WTP was determined by comparing several data obtained from literature studies and evaluation of the existing conditions of Duren Seribu I WTP. Duren Seribu I WTP was evaluated by direct measurement in the field. From the results of data analysis, the design criteria appropriate for Duren Seribu II WTP, the G value is 2078.07 sec-1, the detention time (td) is 4 sec, and the G.td value is 8352.19.

Twin-pipe pumping strategy for stiffening control of 3D printable concrete: From transportation to fabrication

Resolving the contradictions between the pumping and deposition process still remain as a bottleneck in 3D concrete printing. Fluidity retention is beneficial for pumping operations, while fast stiffening after extrusion is indispensable for a high construction rate. To overcome this drawback, an inline mixing strategy, named twin-pipe pumping, using a motionless static mixer and two pumps is proposed. During the printing process, two distinct mixtures are premixed, including a Portland cement-based mixture (without accelerator) and a limestone powder-based mixture (with a high dosage of accelerator). Both mixtures come with a long open time and a high fluidity to optimize the pumping performance. These mixtures are pumped from two separate pipes and merged by the static mixer before extrusion. The stiffening rate of the mixture is then drastically modified inside the static mixer and the combined mixture can reach a high construction rate with good shape stability.

Fully automated microfluidic mixing aided in-line detection of trace level oil field tracers

Recent development in oil field tracer technologies has been focused on implementations that increase oil recovery through optimized algorithms for production, aided by the addition of high fidelity interwell tracer data that improve reservoir history match. Prerequisites for successful tracer campaigns in oil reservoirs at a large scale not only include low-cost tracer materials with a rich palette of uniquely identifiable tracers, but also high sensitivity, easy-to-use trace-level detection and analysis methodologies. A fully automated in-line separation and detection protocol can simultaneously address challenges in throughput (speed and efficiencies) and accuracy (minimization of error-prone steps) of multi-tracer detection. Our previously reported tracer materials consist of barcoded variants of the dipicolinic acid (DPA) molecules that sensitize the emission of lanthanide ions. We have capitalized on the time-gated optical luminescence detectability of this sensitizer-lanthanide ion complex to reject background oil fluorescence, achieving high detection sensitivity in particularly complex oil field produced brine matrix. However, automation of the detection and analysis process has been hindered by opposing requirements in the integration of chromatography instrumentation (separation resolution) with in-line mixing (dispersion due to increase in residence time), and high sensitivity spectrofluorimetry (long integration time). We overcome this major barrier to implementing an in-line detection design under flow conditions by utilizing a microfluidic mixing chip which offers a high degree of mixing in a very short fluidic path length for the in-flow chelation of lanthanide ion to the chromatographically resolved tracer variants, and a flow-cell fluorescence detector at the output of the high-performance liquid chromatography (HPLC). In this study, we demonstrated the rapid in-line detection of a mixture of three different DPA-based barcoded tracers at ultra-trace level. An automated solid phase extraction (SPE) pre-concentration stage is added to the input of the HPLC, further lowering the detection limit down to a level of 100 pM (pico molar) of tracer molecules. As developed, the complete automated system offers unprecedented sensitivity of detection and accelerated oil field tracer analysis that approaches “near real-time” speed.

The effect of different in-line mixers producing emulsifier-free bio-diesel emulsion on the diesel engine combustion performance and exhaust emission

The energy crisis that the world facing right now causes researchers all over the world to opt for alternative fuels. Alternative fuels such as water in diesel emulsion gained attention in various studies due to their performance and capability in reducing harmful emissions such as NOx and PM. A new concept was introduced to make water in diesel emulsion without using a surfactant through a device called Real-Time Non-Surfactant Emulsion Fuel Supply System (RTES) and directly supply the fuel to the engine. This real-time emulsification concept requires a strong mixer, however, it consumes high energy. Thus, this study focused on the effect of different in-line mixers producing emulsifier-free bio-diesel emulsions on stability, diesel engine combustion, performance, and exhaust emission. The original RTES consists of a high shear mixer and ultrasonic mixer, while a new parameter for RTES (RTES-SM) is the high shear mixer and static mixer. The effect of the different inline mixers of RTES and RTES-SM was tested on the stability testing, engine dynamometer under various loading of 2 kW, 4 kW, and 6 kW, and real vehicle testing. The water percentage was set in the range of 7–10 %. Overall, the RTES showed better results on stability and engine dynamometer, but on real vehicle testing, the result is comparable to RTES-SM. Thus, this new concept of an inline mixing system of non-surfactant biodiesel emulsion (RTES-SM) showed better energy efficiency, promising results in real vehicle testing, and potential options in emulsion mixing technologies.

Artificial neural network for mechanism identification and process prediction of the nanoclusters deagglomeration in the in-line HSM

A multiple-input multiple-output artificial neural network (ANN) for the mechanism identification of nanoclusters deagglomeration and the prediction of particle size distribution (PSD) evolution of nanocluster in deagglomeration process of in-line high shear mixer (HSM) was successfully established. It can successfully realize the description of nanoclusters deagglomeration process within single or multiple processing batches. Besides, this ANN model also can realize the accurate prediction of the d32 of fine and coarse agglomerates, and the fines fraction, at different operating conditions and structural parameters, within the relative error of 10 % and 20 % respectively. Meanwhile, 3D profiles of PSD versus different operating conditions and structural parameters were output by the established ANN model, which were useful for parameter selection of in-line HSM to realize efficient production of nanoparticles suspension. The successful establishment of this ANN model provides an effective, flexible, and advanced intelligent method for guiding the parameter selection of in-line HSM, for realizing the controllable and efficient production of the high-end nanoparticle products with specific particle characteristics.

Resolving Solvent Incompatibility in Two-Dimensional Liquid Chromatography with In-Line Mixing Modulation.

Two-dimensional liquid chromatography (2D-LC) is a powerful technique used to characterize complex samples such as synthetic polymers, biomacromolecules, and hybrid modalities (conjugates, oligonucleotides, nanoparticles, etc., which fall between traditional small molecules and large molecules). Characterizing such molecules often requires a highly orthogonal 2D-LC workflow to resolve structurally similar impurities. However, it remains a challenge to achieve truly orthogonal 2D-LC coupling due to incompatibility of the chromatographic conditions used in each dimension. In this work, we present a facile strategy of connecting an in-line mixer, in-line mixing modulation (ILMM), to realize challenging 2D-LC workflows: (1) coupling gel permeation chromatography (GPC) with reversed-phase liquid chromatography (RPLC) for hydrophobic oligomer analysis and (2) coupling ion-pair reversed-phase (IPRP) with hydrophilic interaction liquid chromatography (HILIC) for polar antisense oligonucleotide (ASO) analysis. Compared with the state-of-the-art commercially available active solvent modulation (ASM), engaging the ILMM significantly reduces the peak distortion in GPC-RPLC, allowing an at least 67% higher transfer volume from the primary to secondary dimension, and resolves the ASO sample breakthrough in selective comprehensive IPRP×HILIC. Also remarkably, ILMM demonstrated superiority in comprehensive RPLC×RPLC analysis in comparison with ASM, suggesting its potential in broader 2D-LC applications. In addition to chromatography improvement, ILMM offers several advantages over benchmark modulation approaches in regard to alleviating the need of an additional dilution flow and a simple as well as flexible system configuration, opening many opportunities to establish innovative and versatile multidimensional workflows for characterizing compounds with increasing complexity.

High shear mixer works as a heat exchanger enhancing the liquid–liquid direct contact heat transfer

As an indispensable unit equipment in industrial production, the design of efficient heat exchangers is of great significance to improve energy utilization rate. In this paper, the in-line high shear mixer (HSM) was investigated as a heat exchanger for the first time by experiments and assisted with CFD simulations, and the heat transfer performance was measured using kerosene and thermal water as the heat transfer fluids. Effects of operational parameters, including rotor speed, fluids temperature and flow rate, as well as the structural parameters, involving the rotor teeth number, teeth height, and the shear gap, were studied on the volume heat transfer coefficient and the heat transfer efficiency of such inline HSM. The results indicated that the volume heat transfer coefficient can be significantly improved by increasing the rotor speed and the flow rate of the heat transfer fluids, but reducing the heat transfer temperature difference, whereas increasing the thermal fluid flow rate deteriorates heat transfer efficiency. Furthermore, the structural parameters including rotor teeth number and teeth height, the shear gap within the stator and rotor were optimized to enhance the heat transfer of HSM. The optimal HSM possesses a shear gap of 0.5 mm, a teeth height of 10 mm and a teeth number of 6, which can generate larger turbulent energy dissipation rate and higher shear rate, meanwhile make more fluids jet out of the shear gap and stator hole. The maximum volume heat transfer coefficient and heat transfer efficiency can be up to 2522.8 kW·m−3·°C−1 and 92.4% with the optimal structure. Finally, the dimensionless correlations of heat transfer performance were established to guide the operating mode and structural design of high-efficiency high shear mixer.

Leveraging the Exploratory and Predictive Capabilities of Design of Experiments in Development of Intraarticular Injection of Imatinib Mesylate Containing Lipospheres.

An intraarticular, liposphere-based, formulation of Imatinib mesylate for weekly administration was developed. Lipospheres were prepared using double emulsion technique using dierucoyl phosphatidylcholine, 1,2-dipalmitoyl-sn-glycero-3-phospho-rac-(1-glycerol) sodium salt), cholesterol, and tricaprylin as lipid phase in dichloromethane in a four-step process. Primary emulsion, formed using a high-pressure homogenizer, was diluted using a secondary aqueous phase in an Inline mixer to form the liposomal dispersion. Nitrogen flushing was done to remove dichloromethane, and the dispersion was finally centrifuged and adjusted for potency. The amount of cholesterol and triglyceride was taken as formulation variables, and speed of homogenization was used as a process variable in the Box-Behnken design while particle size, % drug entrapment, and drug release at the end of 4h and 5days were taken as response variables. Multivariate data analysis grouped the variables in two latent variable sets, one based on the speed and the other on the composition of lipospheres. Multiple linear regression analysis was used to generate mathematical model for each response. Constraints were put on the values of responses, as per the requirements of the final product, and the "freedom to operate" design space was located using an overlay plot. The center point batch sufficed all the set criteria, and Monte Carlo simulations on the factor variables indicated a defect rate of 5%. The center point batch was characterized for viscosity, osmolality, pH, drug release, and lipocrit value. The dispersion was charged in a prefilled syringe and studied for stability. The product was found to be stable at 2-8°C over a period of 6months.