Phase Inversion Temperature Method Research Articles

Nanoemulsions encapsulating oregano essential oil: Production, stability, antibacterial activity and incorporation in chicken pâté

Several essential oils (EO) are efficient inhibitors of foodborne pathogen growth. However, their direct incorporation into food presents technological challenges, because of the possibility of extreme changes to their sensory properties. Such drawbacks can be overcome by encapsulation techniques, such as nanoemulsification. This study produced nanoemulsions encapsulating different amounts of oregano (Origanum vulgare) EO, using the phase inversion temperature method, and their physicochemical stability and antibacterial activity were evaluated. The average droplet diameters of the nanoemulsions were in the range of 35–55 nm, and nanoemulsions containing 5 g EO/100 g nanoemulsion were the most stable over the storage period of 45 days. The nanoemulsions exhibited good antibacterial action demonstrated by the low values in vitro for minimum inhibitory concentration (0.56 and 0.60 mg oregano EO/mL for Staphylococcus aureus and Escherichia coli, respectively, and for minimum bactericidal concentration, 0.90 and 3.32 mg oregano EO/mL for Staphylococcus aureus and Escherichia coli, respectively). The nanodispersions also had a higher antibacterial effect against E. coli than for S. aureus. The incorporation of nanoemulsions in chicken pâté did not change the physicochemical characteristics of the meat product. Therefore, the results obtained indicated that nanoemulsions encapsulating oregano EO are suitable to be incorporated into food formulations, such as chicken pâté, to prevent and control microbial growth and to extend their shelf life.

LIPID MICROEMULSION-BASED HYDROGELS FOR EFFECTIVE TOPICAL DELIVERY OF PHENYTOIN

Objective: Microemulsion is a promising drug delivery vehicle for lipophilic drugs but its acceptability for topical application is limited to its very low viscosity. The aim of the present study was to develop and characterize lipid microemulsion hydrogel as a topical drug carrier for phenytoin.Methods: Lipid oil-in-water (O/W) emulsions were formulated from palm kernel oil (PKO), coconut kernel oil (CKO) and soybean oil (SBO), and their blends using phase inversion temperature method. Stable nano-sized microemulsions were identified and formulated into phenytoin loaded hydrogels. The physicochemical properties of the formulations were evaluated in term of emulsion stability index, droplet size, zeta potential, pH, and rheological properties. The efficacy of in vitro drug release of phenytoin was further evaluated using Franz diffusion cells.Results: Stability study revealed that ten lipid emulsions mixing with surfactant Tween 80 at an oil-to-surfactant ratio of 1:9 having 100% emulsion stability indices. Among these, two emulsions (F6 and F21) were identified as the most stable nano-sized microemulsions with clear and transparent appearances; mean droplet size maintained within 100 nm (11–16 nm) as per stability study. Rheological data showed that all phenytoin is loaded hydrogels exhibited non-Newtonian and shear-thinning flow behavior, with high yield stress of a 10.3–18.8 Pa. The in vitro release profiles followed the first-order kinetic model, with R2>0.95, where F21 demonstrated the highest release rate, with 93.12% drug released in 12 h.Conclusion: These findings concluded that CKO/SBO blend microemulsion hydrogel has the highest potential for topical phenytoin delivery.

Manufacture of Oil-in-Water Emulsion Adjuvants.

Emulsion adjuvants for human vaccines have evolved gradually over the last century. Current formulations are the result of many refinements to their composition and manufacturing, as well as optimization for safety and efficacy. Squalene has emerged as being particularly suitable for the manufacturing of safe oil-in-water (O/W) adjuvants for parenteral applications due to its biocompatibility and ability to be metabolized. Emulsion particle size has been identified as an important parameter affecting the pharmaceutical performance of O/W emulsion adjuvants. Submicronic emulsions with sizes in the 80-200 nm range are preferred for potency, manufacturing consistency, and stability reasons. Two manufacturing processes, high pressure homogenization (HPH or microfluidization) and a phase inversion temperature method (PIT), are described to yield such fine and long-term stable emulsion adjuvants.

Antibacterial action of lipid nanocapsules containing fatty acids or monoglycerides as co-surfactants

Lipid nanocapsules (LNCs) are a new generation of biomimetic nanocarriers obtained via a phase inversion temperature method and have an oily core of medium-chain triglycerides that is surrounded by a shelf containing a lipophilic surfactant (lecithin) and a hydrophilic surfactant macrogol 15-hydroxystearate. The aim of the present study was to produce LNCs with antibacterial activity by replacing lecithin with other lipophilic surface active compounds, namely medium-chain fatty acids and their 1-monoglycerides, which are known to have antimicrobial properties.Fatty acids and monoglycerides were found to affect the properties of LNCs, such as particle size and zeta potential. Incorporation of a co-surfactant decreased significantly particle size (p⩽0.0039). Furthermore, incorporation of either lecithin or fatty acids with at least 10 carbon atoms yielded LNCs with the zeta potential significantly more negative than that of LNCs composed solely of triglycerides and macrogol 15 hydroxystearate (p⩽0.0310). Moreover, they were capable of decreasing the phase inversion temperature. The activity of the LCNs against Gram-positive S. aureus, including a methicillin-resistant strain, increased with increases in the length of the hydrocarbon tail. Monoglyceride-LNCs were found to be more active than the corresponding fatty acids. The opposite behaviour was observed for Gram-negative bacteria, whereby only caproic acid- and caprylic acid-LNCs were found to be active against these organisms. The monoglyceride-LNCs were bactericidal, and they killed in a time-dependent manner. Fatty acid-LNCs killed in a concentration-dependent manner.A haemolysis assay was performed to obtain preliminary information on the safety of the tested LNCs. In the case of fatty acid-LNCs, the concentrations at which bacterial growth was inhibited were similar to the haemolytic concentrations. However, monoglyceride-LNCs showed antibacterial action at concentrations much lower than those at which haemolysis was observed. In conclusion, monoglyceride-LNCs are promising candidates as carriers for the encapsulation of antibacterial agents, particularly against S. aureus.

Physico-chemical stability and in vitro digestibility of beta-carotene-loaded lipid nanoparticles of cupuacu butter (Theobroma grandiflorum) produced by the phase inversion temperature (PIT) method

Beta-carotene is a carotenoid with a wide spectrum of biological activities (e.g., anti-cancer, anti-hypertensive, and anti-inflammatory). However, because of its extremely high hydrophobicity, it is difficult to incorporate in food formulations and its bioavailability is fairly low. Lipid-based encapsulation colloidal systems such as lipid nanoparticles can help overcome these issues. In this study, beta-carotene-loaded lipid nanoparticles were produced by the phase inversion temperature (PIT) method from 10% cupuacu butter and 20% surfactant (Cremophor RH40 and Span 80). The inversion temperature of the nanoparticles was 74 °C and their average diameter was 35 nm. After 100 days of storage, 85% of the initial amount of beta-carotene remained in the nanoparticles; alpha-tocopherol was found to be essential for carotenoid preservation. Comparison of the results of in vitro digestion between static and dynamic systems was performed, and the characteristics of each digestion system led to diverse results in terms of average particle size and beta-carotene bioaccessibility. Although the static system was much simpler than the dynamic system, it could not provide reliable data of the digestibility of the lipid nanoparticles. The bioaccessibility of beta-carotene in the static system was 92%, very similar to the results found in the literature; by comparison, the dynamic system revealed a beta-carotene bioaccessibility of nearly 20%. Despite this discrepancy, the highly realistic conditions of digestion simulated by the dynamic in vitro system indicate that the results of this system are more reliable than those obtained from the simplified static system applied in this research.

Effect of lipid nanoparticle formulations on skin delivery of a lipophilic substance

The aim of this study was to follow the skin penetration of a model lipophilic compound (Nile red) delivered by nanoparticulate carriers, the so-called lipid nanocapsules. The nanocapsules consisting of an oil core stabilized by amixture of surfactants were prepared by the phase inversion temperature method. Varying the particle composition (the oil/surfactant ratio) nanoparticles of different size were prepared and characterized. The penetration profile of Nile red delivered into the porcine skin by the nanoparticles compared to non-particulate samples was determined using fluorescence microscopy combined with a novel, statistically robust quantitative image analysis method. This study demonstrated that lipid nanoparticles promoted the skin penetration of encapsulated Nile red in comparison with all the non-particulate samples. Nile red delivered by the lipid-based nanoparticles was able to diffuse across the stratum corneum and partition itself uniformly in the epidermis. No relationship between Nile red penetration into the skin and the particle size was found. Moreover, the presence of sodium chloride in the water phase had a negative impact on the Nile red penetration into the skin. The results indicate that the physico-chemical circumstances of the nanoparticulate formulation play the major role in the penetration of lipophilic substances into the skin.

Formation and stability of solid lipid nanoparticles fabricated using phase inversion temperature method

Abstract Solid lipid nanoparticles (SLNs) are being investigated for their ability to encapsulate and protect lipophilic bioactive compounds in foods, supplements, and pharmaceuticals. In this study, the phase inversion temperature (PIT) method was used to fabricate SLNs using a model surfactant (Brij 30, C12E4)/oil (octadecane)/water system. Surfactant/oil/water (SOW) mixtures were maintained at a temperature above the PIT, and then rapidly cooled to a temperature below the lipid nanoparticle crystallization point. The PIT (≈40 °C) was determined by monitoring the turbidity versus temperature profile of the SOW system during heating. The lipid nanoparticle crystallization point, melting point, and physical state were determined using differential scanning calorimetry (DSC). The stability of the lipid nanoparticles after fabrication depended on the storage temperature relative to the PIT and melting/crystallization points. At temperatures appreciably below their melting point (≈26 °C), the lipid nanoparticles were completely solid and stable to aggregation. At temperatures around their melting point, the lipid nanoparticles were partially crystalline, which led to partial coalescence and gelation. At temperatures appreciably above their melting point but below their PIT, the lipid nanoparticles were completely liquid and prone to coalescence and phase separation. These results have important implications for optimizing the fabrication and storage conditions required to produce stable nanoemulsions suitable for utilization in commercial products using low-energy methods.

Liquid crystal nanoparticle formulation as an oral drug delivery system for liver-specific distribution.

Liquid crystal nanoparticles have been utilized as an efficient tool for drug delivery with enhanced bioavailability, drug stability, and targeted drug delivery. However, the high energy requirements and the high cost of the liquid crystal preparation have been obstacles to their widespread use in the pharmaceutical industry. In this study, we prepared liquid crystal nanoparticles using a phase-inversion temperature method, which is a uniquely low energy process. Particles prepared with the above method were estimated to be ~100 nm in size and exhibited a lamellar liquid crystal structure with orthorhombic lateral packing. Pharmacokinetic and tissue distribution studies of a hydrophobic peptide-based drug candidate formulated with the liquid crystal nanoparticles showed a five-fold enhancement of bioavailability, sustained release, and liver-specific drug delivery compared to a host–guest complex formulation.

Oral Delivery of a High Quercetin Payload Nanosized Emulsion: In Vitro and In Vivo Activity Against B16-F10 Melanoma.

Quercetin is a natural compound that has several biological activities including anticancer activity. However, the use of this drug has been limited mostly because of its poor water solubility and low bioavailability. Therefore, the development of quercetin-loaded nanocarrier systems may be considered a promising advance to exploit its therapeutic properties in clinical setting including cancer treatment. This study evaluates the effect of orally administered nanosized emulsion containing quercetin (QU-NE) on the cytotoxicity activity against B16-F10 cells in vitro, and on subcutaneous melanoma in mice inoculated with B16-F1O cells. In vivo experiments, also evaluate the co-administration of quercetin with cisplatin in order to predict synergic effects and the renal and hepatic toxicity. The nanocarriers were prepared through the hot solvent diffusion associated with the phase inversion temperature methods. In vitro study showed reduction of cell viability in a concentration-depend manner for free quercetin and QU-NE. In vivo study, quercetin either as a free drug or colloidal dispersion was administrated at a dose of 5 mg kg(-1) twice a week for 17 days via oral route. Cisplatin was administrated at dose of 1 mg kg(-1) once a week intraperitoneally. Free quercetin and QU-NE reduced tumor growth, however, the reduction observed for QU-NE (P < 0.001 vs. control) was significantly higher than free quercetin (P < 0.05 vs. control). The association of both drugs did not show synergic effect. Besides, no renal or hepatic toxicities were observed after administration of free quercetin and QU-NE. These results suggest that an improvement in the oral bioavailability of quercetin occurred when this compound was dissolved in the oily phase of a nanosized emulsion, indicating that it might have a potential application in the treatment of melanoma.

The effect of concentration of surfactant and electrolyte on the pit and droplet sizes nanoemulsions of n-dodecane in water

Nanoemulsions are transparent or semi-transparent systems mostly covering the size range of 50–200 nm. In the research, the production of nanoemulsion of n-dodecane in water was studied by phase inversion temperature method (PIT), and the effect of the changes in NaCl and surfactant concentration on PIT and droplet size of n-dodecane was studied. The changes in conductivity with temperature were measured for emulsions containing 20 wt % dodecane, 4 and 8 wt % Brij30, and NaCl of various concentrations in aqueous phase. The results showed a decrease in PIT with a rise in concentration of NaCl and also Brij30. There was a linear model fitted to the data. Analysis of variance (ANOVA) shows that, the significance value (p-value) is less than 0.05, and the effect of the changes in concentration of surfactant Brij30 on the decrease in PIT and droplet size was the more than that in concentration of NaCl in aqueous phase. The lowest mean droplet diameters were in the range of 8.6–14 nm.

Anti-inflammatory Effect and Toxicology Analysis of Oral Delivery Quercetin Nanosized Emulsion in Rats

This study evaluates the advantage of the quercetin encapsulation in nanosized emulsion (QU-NE) administered orally in rats in order to demonstrate its anti-oedematous and antioxidant effects as well as its toxicity. The nanocarriers were prepared using the hot solvent diffusion with the phase inversion temperature methods. The nanocarriers physicochemical properties were then investigated. The anti-edematous activity was tested using paw edema in rats. In addition, NF-kB expression in subcutaneous tissue of the paws was accessed by immunohistochemistry while the lipid peroxidation was analyzed in the liver by malondialdehyde reaction with thiobarbituric acid. Hematological, renal and hepatic toxicity as well as the genetic damage were also evaluated. The results demonstrated that QU-NE exhibited pronounced anti-oedematous property comparable to drug diclofenac. This effect was associated with NF-κB pathway inhibition. The lipid peroxidation was also only reduced in rats treated with QU-NE. Besides this, no genetic damage, hematological, renal or hepatic toxicities were observed after administration of QU-NE. These results suggest that quercetin nanosized emulsion exhibits anti-oedematous and antioxidant properties and does not demonstrate toxic effects. This indicates that it has a potential application in the treatment of inflammatory diseases.

Solid Lipid Nanoparticles (SLNs) for Intracellular Targeting Applications.

Nanoparticle-based delivery vehicles have shown great promise for intracellular targeting applications, providing a mechanism to specifically alter cellular signaling and gene expression. In a previous investigation, the synthesis of ultra-small solid lipid nanoparticles (SLNs) for topical drug delivery and biomarker detection applications was demonstrated. SLNs are a well-studied example of a nanoparticle delivery system that has emerged as a promising drug delivery vehicle. In this study, SLNs were loaded with a fluorescent dye and used as a model to investigate particle-cell interactions. The phase inversion temperature (PIT) method was used for the synthesis of ultra-small populations of biocompatible nanoparticles. A 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenylphenyltetrazolium bromide (MTT) assay was utilized in order to establish appropriate dosing levels prior to the nanoparticle-cell interaction studies. Furthermore, primary human dermal fibroblasts and mouse dendritic cells were exposed to dye-loaded SLN over time and the interactions with respect to toxicity and particle uptake were characterized using fluorescence microscopy and flow cytometry. This study demonstrated that ultra-small SLNs, as a nanoparticle delivery system, are suitable for intracellular targeting of different cell types.

Solid Lipid Nanoparticles (SLNs) for Intracellular Targeting Applications

Nanoparticle-based delivery vehicles have shown great promise for intracellular targeting applications, providing a mechanism to specifically alter cellular signaling and gene expression. In a previous investigation, the synthesis of ultra-small solid lipid nanoparticles (SLNs) for topical drug delivery and biomarker detection applications was demonstrated. SLNs are a well-studied example of a nanoparticle delivery system that has emerged as a promising drug delivery vehicle. In this study, SLNs were loaded with a fluorescent dye and used as a model to investigate particle-cell interactions. The phase inversion temperature (PIT) method was used for the synthesis of ultra-small populations of biocompatible nanoparticles. A 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenylphenyltetrazolium bromide (MTT) assay was utilized in order to establish appropriate dosing levels prior to the nanoparticle-cell interaction studies. Furthermore, primary human dermal fibroblasts and mouse dendritic cells were exposed to dye-loaded SLN over time and the interactions with respect to toxicity and particle uptake were characterized using fluorescence microscopy and flow cytometry. This study demonstrated that ultra-small SLNs, as a nanoparticle delivery system, are suitable for intracellular targeting of different cell types.

Lemon oil nanoemulsions fabricated with sodium caseinate and Tween 20 using phase inversion temperature method

The phase inversion temperature (PIT) method has been studied to fabricate nanoemulsions using non-ionic synthetic surfactants, but not food biopolymers. The objective of this study was to investigate the PIT formation of lemon oil nanoemulsions by combinations of Tween 20 and sodium caseinate (NaCas). The emulsions prepared with combinations of NaCas and Tween 20 had lower turbidity and smaller droplets than those prepared with one surfactant, and the co-adsorption of NaCas on oil droplets decreased with increasing Tween 20 concentration. Negative and positive impacts on nanoemulsion formation were observed at 0.2–0.4 and 0.6–0.8 mM NaCl, respectively. Turbidity and rheology results showed the PIT between 80 and 90 °C. The nanoemulsions prepared with 2% NaCas, 0.4–1.2% Tween 20 and 1.5% lemon oil had a volume-area mean diameter of around 100 nm and were stable during 15-day storage. Therefore, NaCas can be used to partially replace synthetic surfactants to prepare flavor oils nanoemulsions using the PIT method.

Fabrication of protein nanoparticles and microparticles within water domains formed in surfactant–oil–water mixtures: Phase inversion temperature method

Protein nanoparticles and microparticles can be formed using numerous fabrication methods. In this study, a templating method was developed that involves trapping globular proteins within the aqueous domains formed by surfactant–oil–water (SOW) mixtures, and then inducing protein aggregation and particle formation through thermal denaturation. A heat-set gelling protein (whey protein isolate) was located in the aqueous phase of an oil-in-water nanoemulsion formed by spontaneous emulsification. This system was then heated to promote the conversion of the oil-in-water nanoemulsion into a bicontinuous microemulsion at the phase inversion temperature (PIT) or a water-in-oil emulsion above the PIT. In principle, the nature of the aqueous domains present in the SOW system when the protein molecules unfold and aggregate determine the size and morphology of the protein particles formed. In this study, turbidity, rheology, and confocal fluorescence microscopy measurements were used to show that spheroid protein particles (1–10 μm) could be formed by heating SOW mixtures to a temperature above the PIT (which led to the formation of W/O emulsions) and where protein denaturation and aggregation occurred. This approach offers an alternative to existing methods of fabricating protein nanoparticles and microparticles suitable for utilization in the food and other industries. The major disadvantages of the method are that appreciable amounts of surfactant and oil are required in the particle-templating process, and that elevated temperatures are required to induce phase inversion and thermal denaturation.

Detailed investigation of nano-emulsions obtained from the Remcopal 4/decane/water system

Abstract We have investigated the phase behavior of the Remcopal 4/decane/water system. Direct lamellar phase (L α ), reverse hexagonal (H 2 ) liquid crystals, and reverse micellar phase L 2 were identified. Then, nano-emulsions have been formulated by the phase inversion temperature (PIT) method and characterized by various techniques such as dynamic light scattering (DLS), nanoparticle tracking analysis (NTA), conductivity, and small angle X-ray scattering (SAXS). For oil/surfactant ratios comprising between 2.3 and 5.7, a domain of nano-emulsion, stable between 20 and 27 °C, which extend up to 95 wt.% of water, is formed. The size of nano-emulsions is around 100 nm of diameters. It increases with the oil/surfactant ratio. The conductivity and the SAXS analyzes show that there is a strong relation between the joint presence of a dispersion of the lamellar phase L α , the sponge phase L3, and the formation of the nano-emulsions. Finally, the obtained nano-emulsions have been used as building blocks to prepare porous silica. Surprisingly, materials with a pore size in the micrometer range have been recovered.

Preparation of Virgin Coconut Oil Nanoemulsions by Phase Inversion Temperature Method

Presently, the low energy emulsification method; phase inversion temperature (PIT) method offers a convenient mean of nanoemulsion formation for the incorporation of fragile substances. The aim of the current study was to prepare coconut oil nanoemulsions using cremophor RH 40 as nonionic surfactant in the light of PIT method. The suitable composition of coconut oil, cremophor RH40 and water was investigated by construction of ternary phase diagram. The PIT of system can be adjusted to 75 °C by adding NaCl at the concentration of 6%(w/w). Stable translucent nanoemulsions with particle size in range of 20-100 nm and PDI&lt;0.2 can be formed at the constituent proportions of 5%-30%(w/w), coconut oil, 10%-80%(w/w) cremophor RH40 and 5%-85%(w/w) NaCl solution of 6%(w/w). All formulations were stable for up to 2 months when stored at 4, 30 and 40 °C and protected from light.

Idebenone loaded solid lipid nanoparticles: calorimetric studies on surfactant and drug loading effects.

In this study we prepared solid lipid nanoparticles (SLN), by the phase inversion temperature (PIT) method, using cetyl palmitate as solid lipid and three different non-ionic emulsifiers of the polyoxyethylene ethers family (ceteth-20, isoceteth-20, oleth-20). These SLN were loaded with different amount of idebenone (IDE), an antioxidant drug useful in the treatment of neurodegenerative diseases and skin oxidative damages. The differential scanning calorimetry (DSC) was employed to evaluate the effects of the different emulsifiers and the different amounts of drug loaded on the thermotropic behavior of SLN and to investigate how the drug was arranged into these nanoparticles. The IDE seemed to be located into different regions of the SLN depending on its concentration and on the surfactant used. The results of this study suggest that the calorimetric studies performed on SLN could provide valuable information to optimize SLN design and drug release from these carriers.

PIT tuning effects of hydrophobic co-surfactants and drugs

The formation of O/W nanoemulsions by the phase inversion temperature (PIT) method has been studied in water/Miglyol 812 systems. The effects of a hydrophobic co-surfactant (Igepal) and a hydrophobic drug (α-tocopherol) on tuning and optimising the PIT, which is essential for industrial processes, were studied. The hydrophilic poly-oxyethylated surfactant used was Kolliphor HS15. For these systems temperature induced phase inversion was not feasible in the presence of the hydrophilic surfactant alone. The addition of the hydrophobic co-surfactant was necessary to induce phase inversion and formation of stable O/W emulsions. Increasing the concentration of either the hydrophobic surfactant or of the hydrophobic drug, led to the formation of smaller drops, to the decrease of the PIT and the decrease of the transitional phase inversion zone. The effects of the hydrophobic/hydrophilic surfactant ratio, and of the hydrophobic drug concentration on tuning and optimising the PIT and the drop diameter were studied. Nanoemulsions with mean drop diameter ranging from 80nm to 196nm were produced for Kolliphor/Igepal systems and from 50nm to 100nm for Kolliphor/α-tocopherol systems. The polydispersity index was smaller than 0.2. α-Tocopherol acted as a co-surfactant. It could replace the hydrophobic co-surfactant, Igepal, and led to the decrease of the PIT by 10–15°C. Temperature cycling did not have any significant effect on the PIT, nor on the drop sizes. However, it had a significant effect on the phase inversion process by changing the phase inversion path.

Synthesis of sub-10 nm solid lipid nanoparticles for topical and biomarker detection applications

Solid lipid nanoparticles (SLNs) are a promising platform for sensing in vivo biomarkers due to their biocompatibility, stability, and their ability to carry a wide range of active ingredients. The skin is a prominent target organ for numerous inflammatory and stress-related biomarkers, making it an excellent site for early detection of physiological imbalance and application of sensory nanoparticles. Though smaller particle size has generally been correlated with increased penetration of skin models, there has been little attention paid to the significance of other nanoparticle synthesis parameters with respect to their physical properties. In this study, we demonstrate the synthesis of sub-10 nm SLNs by the phase inversion temperature (PIT) method. These particles were specifically designed for topical delivery of hydrogen peroxide-detecting chemiluminescent dyes. A systematic design of experiments approach was used to investigate the role of the processing variables on SLN form and properties. The processing variables were correlated with the SLN properties (e.g., dye solubility, phase inversion temperature, particle size, polydispersity, melting point, and latent heat of melting). Statistical analysis revealed that the PIT method, while allowing total control over the thermal properties, resulted in well-controlled synthesis of ultra-small particles, while allowing great flexibility in the processing conditions and incorporated compounds.