High Pressure Microfluidization Research Articles

Food-grade emulsion gels and oleogels prepared by all-natural dual nanofibril system from citrus fiber and glycyrrhizic acid

The natural dual nanofibril system consisting of the rigid semicrystalline nanofibrils disintegrated from citrus fiber (CF) and soft semiflexible nanofibrils self-assembled from glycyrrhizic acid (GA) has been recently shown to be effective structural building blocks for fabrication of emulsion gels. In this work, the effect of the CF nanofibrils prepared by different mechanical disintegration approaches (i.e., high-pressure microfluidization and hydrodynamic cavitation) on the interfibrillar CF-GA interactions and the subsequent formation and properties of emulsion gels were investigated, with the aim of evaluating the potential of the dual nanofibril-stabilized emulsion gels as templates for synthesizing all-natural edible oleogels. The obtained results demonstrate that compared to the cavitation, the high-pressure microfluidization is more capable of generating CF nanofibrils with a higher degree of nanofibrillation and individualization, thus forming a denser CF-GA gel network with higher viscoelasticity and structural stability due to the stronger multiple intrafibrillar and interfibrillar interactions. The emulsion gels stabilized by the dual nanofibril system are demonstrated to be an efficient template to fabricate solid-like oleogels, and the structural properties of the oleogels can be well tuned by the mechanical disintegration of CF and the GA nanofibril concentration. The prepared oleogels possess high oil loading capacity, dense network microstructure, superior rheological and large deformation compression performances, and satisfactory thermal stability, which is attributed to the compact and ordered CF-GA dual nanofibrillar network via multiple hydrogen-bonding interactions in the continuous phase as well as at the droplet surface. This study highlights the unique use of all-natural dual nanofibrils to develop oil structured soft materials for sustainable applications.

Sub-micron colloidosomes with tuneable cargo release prepared using epoxy-functional diblock copolymer nanoparticles

HypothesisPickering emulsions stabilized using epoxy-functional block copolymer nanoparticles should enable the formation of sub-micron colloidosomes that are stable with respect to Ostwald ripening and allow tuneable small-molecule cargo release. ExperimentsEpoxy-functional diblock copolymer nanoparticles of 24 ± 4 nm were prepared via reversible addition-fragmentation chain transfer (RAFT)-mediated dispersion polymerization of methyl methacrylate (MMA) in n-dodecane. Sub-micron water-in-n-dodecane Pickering emulsions were prepared by high-pressure microfluidization. The epoxy groups were then ring-opened using 3-aminopropyltriethoxysilane (APTES) to prepare cross-linked colloidosomes. The colloidosomes survived removal of the aqueous phase using excess solvent. The silica shell thickness could be adjusted from 11 to 23 nm by varying the APTES/GlyMA molar ratio. The long-term stability of the colloidosomes was compared to precursor Pickering emulsions. Finally, the permeability of the colloidosomes was examined by encapsulation and release of a small molecule. FindingsThe Pickering emulsion droplet diameter was reduced from 700 to 200 nm by increasing the salt concentration within the aqueous phase. In the absence of salt, emulsion droplets were unstable due to Ostwald ripening. However, emulsions prepared with 0.5 M NaCl are stable for at least one month. The cross-linked colloidosomes demonstrated much more stable than the precursor sub-micron emulsions prepared without salt. The precursor nanoemulsions exhibited complete release (>99 %) of an encapsulated dye, while higher APTES/GlyMA ratios resulted in much lower dye release, yielding nearly impermeable silica capsules that retained around 95 % of the dye.

Effect of Dynamic High-Pressure Microfluidization on the Quality of Not-from-Concentrate Cucumber Juice.

The effects of dynamic high-pressure microfluidization (DHPM at 400 MPa) and heat treatment (HT) on the microbial inactivation, quality parameters, and flavor components of not-from-concentrate (NFC) cucumber juice were investigated. Total aerobic bacteria, yeasts and molds were not detected in the 400 MPa-treated cucumber juice. Total phenolic content increased by 16.2% in the 400 MPa-treated cucumber juice compared to the control check (CK). The significant reduction in pulp particle size (volume peak decreasing from 100-1000 μm to 10-100 μm) and viscosity increased the stability of the cucumber juice while decreasing the fluid resistance during processing. HT decreased the ascorbic acid content by 25.9% (p < 0.05), while the decrease in ascorbic acid content was not significant after 400 MPa treatment. A total of 59 volatile aroma substances were identified by gas chromatography-ion mobility spectrometry (GC-IMS), and a variety of characteristic aroma substances (i.e., valeraldehyde, (E)-2-hexenal, (E)-2-nonenal, and (E,Z)-2,6-nonadienal, among others) were retained after treatment with 400 MPa. In this study, DHPM technology was innovatively applied to cucumber juice processing with the aim of providing a continuous non-thermal processing technology for the industrial production of cucumber juice. Our results provide a theoretical basis for the application of DHPM technology in cucumber juice production.

Dynamic high pressure microfluidization modified oat dietary fiber: Texture modulation and its mechanistic in whole grain oat milk

Oat dietary fiber (ODF) could product some undesirable textures such as rough and thick when added to food products, resulting the waste of resources. In this study, dynamic high pressure microfluidization (DHPM) was used to treat ODF in different pressures, and ODF was backfilled into whole grain oat milk (WOM) to investigate the efficiency of DHPM in improving the undesirable taste from ODF. The physicochemical properties and the microstructure of the DHPM-treated ODF were also determined to investigate the modification mechanism. The results showed that the WOM with DHPM-treated ODF conferred lower particle size (D[90] decreasing by 22.80%–51.35%), firmness (decreasing by 0.08%–9.84%), apparent viscosity and coefficient of friction, especially 120 and 150 MPa. Meanwhile, the samples with DHPM had higher sensory scores such as particles detected (swallowing and after swallowing increasing by 38.10%–85.71% and 29.17%–62.50% respectively), smoothness (increasing by 6.9%–27.59%), and mouth-coating, indicating that DHPM treatment could improve the undesirable texture brought by ODF. In addition, DHPM treatment reduced the particle size and insoluble dietary fiber content and fragmented microstructure of ODF, it may weaken the particles detected and improve the smooth taste. The consistency coefficient of ODF decreased from 0.0091 (0 MPa) to 0.0038 (150 MPa), which could be related to the improvement of texture flow and mouth-coating. In conclusion, WOM with DHPM-treatment ODF improved the undesirable texture such as roughness and stickiness, possibly associated with the ability of DHPM to alter the particle size, apparent viscosity and microstructure of ODF. This study provided the method of modification and utilization of ODF, and provided the idea and feasible solution for preparation of WOM.

Fabrication of cinnamon essential oil nanoemulsions with high antibacterial activities via microfluidization

The high volatility and hydrophobicity of cinnamon essential oils (CiEO) limited their practical application. To enhance their stability and antibacterial activity, nanoemulsions encapsulating CiEO were prepared using hydroxypropyl-β-cyclodextrin/lauroyl arginate (HPCD/LAE) inclusion complexes through high-pressure microfluidization (HPM). Effects of HPM parameters on the stability and antibacterial properties of nanoemulsion were investigated. Results revealed that increased processing pressure and cycle numbers were associated with reduced droplet size and greater homogeneity in CiEO distribution. Storage and thermal stability were optimized at 100 MPa and seven cycles. Moreover, the nanoemulsions showed strong synergistic antibacterial against E. coli (19.79 mm) and S. aureus (23.61 mm) compared with LAE (11.52 mm and 12.82 mm, respectively) and CiEO alone (13.26 mm and 17.68 mm, respectively). This study provided new information for constructing CiEO nanoemulsion, which is suitable for use in the food industry.

Preparation of High-Toughness Cellulose Nanofiber/Polylactic Acid Bionanocomposite Films via Gel-like Cellulose Nanofibers.

This study demonstrates a procedure for preparing gel-like cellulose nanofibers (CNFs) in polyethylene glycol (PEG) to toughen polylactic acid (PLA) nanocomposite films. A well-dispersed solution of CNFs in ethanol was produced from microcrystalline cellulose by using a high-pressure microfluidizer. The fiber diameter of CNFs was found to be in the range of 80-100 nm. Ethanol was replaced by PEG using a rotary evaporator to obtain gel-like CNFs/PEG. PLA/PEG/CNF films were prepared using the solvent casting method, with the CNF content varying from 0.15 to 5 phr. The effect of CNFs on the mechanical, morphological, and thermal properties of PLA nanocomposite films was investigated. The results demonstrate that the addition of CNFs improved Young's modulus and toughness of PLA/PEG films. In contrast, a slight decrease in mechanical properties was observed when the content of CNFs reached 0.83 phr. Considère's constructions are used to explain the neck phenomena and cold drawing of nanocomposite films. The crystallization and thermal stability of PLA nanocomposite films were enhanced, with a slight decrease in cold-crystalline temperature (T cc) and an increase in decomposition temperature (T d).

Rheological properties, multiscale structure, and in vitro digestibility of a maize starch–konjac glucomannan–bamboo leaf flavonoid complex modified by dynamic high-pressure microfluidization

The effects of dynamic high-pressure microfluidization (DHPM) treatment on the rheological properties, multiscale structure and in vitro digestibility of complex of maize starch (MS), konjac glucomannan (KGM), and bamboo leaf flavonoids (BLFs) were investigated. Compared with MS, the MS–KGM–BLF complex exhibited reduced viscosity and crystallinity, along with increased lamellar thickness to 10.26 nm. MS–KGM–BLF complex had lower viscosity after DHPM treatment. The highest ordered structure and crystallinity were observed at 50 MPa, with the α value increasing from 3.40 to 3.59 and the d value decreasing from 10.26 to 9.81 nm. However, higher DHPM pressures resulted in a decrease in the α value and an increase in the d value. The highest gelatinization enthalpy and resistant starch content were achieved at 100 MPa DHPM, while the fractal structure shifted from surface fractal to mass fractal at 150 MPa. This study presents an innovative method for enhancing the properties of MS.

Comparative study on dynamic in vitro digestion characteristics of lotus seed starch-EGCG complex prepared by different processing methods

To study the effect of starch-polyphenol interaction induced by different processing methods on digestion characteristics, a dynamic in vitro human gastrointestinal system was employed to investigate the digestive characteristics of lotus seed starch–epigallocatechin gallate (EGCG) complex (LS–EGCG) prepared by different processing methods. Digestion altered crystal structure, particle size, morphology, pH, starch hydrolysis, and EGCG content. Processing broke physical barriers, reducing particle size by enzyme erosion. Enzymatic hydrolysis gradually exposed EGCG, indicated by green fluorescence. Heat and high pressure treatments enhanced starch dissolution, increasing sugar accumulation and hydrolysis. However, ultrasonic-microwave and high pressure microfluidization treatments formed dense structures, decreasing hydrolysis rates. Overall, the complex formed by high pressure microfluidization showed better enzyme resistance. The results provide a scientific basis for the development of food with quality and functional properties.

Enhancing the usability of pea protein in emulsion applications through modification by various approaches: A comparative study

The extensive utilization in food industry of pea protein is often impeded by its low water solubility, resulting in poor functional properties. Various methods, including pH-shifting (PS), ultrasonication (US), high-pressure micro-fluidization (MF), pH-shifting combined with ultrasonication (PS-US), and pH-shifting with micro-fluidization (PS-MF), were utilized to modify pea protein isolate (PPI) in order to enhance its functionality in emulsion formulation. The physicochemical properties and structural changes of the protein were investigated by assessing solubility, particle size, surface charge, protein profile, surface hydrophobicity, free sulfhydryl groups, and secondary structure content. The extent of modification induced by each treatment method on PPI-stabilized emulsions was compared based on parameters such as adsorbed interfacial protein concentration, particle size, zeta potential, and microstructure of the prepared emulsions. All modification increased the solubility of pea protein in the sequence of PS (4-fold) < MF (7-fold) < US (11-fold) < PS-US (13-fold) < PS-MF (14-fold). For single treatments, proteins dissolved more readily under US, resulting in the most uniform emulsions with small particle. The combined processes of PS-US and PS-MF further improved solubility, decreased emulsions particle size, promoted uniformity of emulsions. PS-US-stabilized emulsions displayed more smaller droplet size, narrower size distribution, and slightly higher stability than those prepared by PS-MF. The relatively higher emulsifying capacity of PPI treated by PS-US than those by PS-MF may be attributed to its higher surface hydrophobicity.

Perilla protein isolate exhibits synergistic techno-functionality through modification via sequential dynamic high-pressure microfluidization and enzymatic hydrolysis

Modification of proteins derived from plant sources is crucial for enabling their versatile applications in the food industry. In the present study, structural and functional properties of individual treated perilla protein isolate (PPI) through dynamic high-pressure microfluidization (DHPM) and enzymatic hydrolysis (EH) was compared with their sequential (DHPM-EH) treatment. Native PPI thick, sharp-edged blocks transformed into a collapsed structure with wrinkles after sequential (DHPM-EH) treatment. The synergistic effect between DHPM and EH, during sequential (DHPM-EH) treatment caused greater reduction (91.9%) in PPI particle size as compared to DHPM (87.3%), and EH (84.5%), which enhanced PPI solubility, even at low pH. Above treatments altered the conformational structure of PPI as evident from changes in surface hydrophobicity, free –SH groups conversion into –SS bonds, ultraviolet absorption and fluorescence intensities changes, and decreased α-helix, while increased β-sheets. The sequential (DHPM-EH) treated PPI showed enhanced foaming capacity (×3.59), as well as increased thermal and colloidal stability.

Effect of dynamic high-pressure microfluidization on structural, physicochemical, and functional properties of pea albumin

Effect of dynamic high-pressure microfluidization on structural, physicochemical, and functional properties of pea albumin

Gastrointestinal digestion behavior and bioavailability of greenly prepared highly loaded myofibrillar-luteolin vehicle

In this study, the highly loaded myofibrillar protein (MP)-luteolin (Lut) complexes were noncovalently constructed by using green high-pressure homogenization technology (HPH) and high-pressure micro-fluidization technology (HPM), aiming to optimize the encapsulation efficiency of flavonoids in the protein-based vehicle without relying on the organic solvent (i.e. DMSO, ethanol, etc.). The loading efficiency of Lut into MPs could reach 100 % with a concentration of 120 μmol/g protein by using HPH (103 MPa, 2 passes) without ethanol adoption. The in vitro gastrointestinal digestion behavior and antioxidant activity of the complexes were then compared with those of ethanol-assisted groups. During gastrointestinal digestion, the MP digestibility of complexes, reaching more than 70.56 % after thermal treatment, was higher than that of sole protein. The release profile of Lut encapsulated in ethanol-containing and ethanol-free samples both well fitted with the Hixson-Crowell release kinetic model (R2 = 0.92 and 0.94, respectively), and the total phenol content decreased by ≥ 40.02 % and ≥ 62.62 %, respectively. The in vitro antioxidant activity (DPPH, ABTS, and Fe2+) of the digestive products was significantly improved by 23.89 %, 159.69 %, 351.12 % (ethanol groups) and 13.43 %, 125.48 %, 213.95 % (non-ethanol groups). The 3 mg/mL freeze-dried digesta significantly alleviated lipid accumulation and oxidative stress in HepG2 cells. The triglycerides and malondialdehyde contents decreased by at least 57.62 % and 67.74 % after digesta treatment. This study provides an easily approached and environment-friendly strategy to construct a highly loaded protein–flavonoid conjugate, which showed great potential in the formulation of healthier meat products.

Development of chia oil-in-water nanoemulsions using different homogenization technologies and the layer-by-layer technique

Aim: The present study investigates the influence of various homogenization techniques, namely high-pressure valve homogenization and microfluidization, and different forms of modified sunflower lecithin, including deoiled (DL) and hydrolyzed (HL) variants, on the development of monolayer and bilayer nanoemulsions of chia oil. Methods: Oil-in-water (O/W) nanoemulsions with 5% chia seed oil were prepared using simple (0.5% DL or HL) or double-layer [0.5% DL or HL and 0.3% chitosan (Ch)] stabilization. This involved a two-step homogenization process, utilizing either microfluidization or high-pressure valve homogenization. Chia oil nanoemulsions were characterized by their zeta potential, particle size, and rheological properties. Besides, their physical stability and omega-3 content during refrigerated storage were evaluated. Results: Overall, the studied modified sunflower lecithin (DL and HL) demonstrated effective capability in stabilizing chia nanoemulsions and facilitating the formation of the double-layered structure following Ch deposition. Concerning the homogenization method, it has been demonstrated that under the same homogenization conditions, microfluidization resulted in significantly smaller droplet sizes and higher apparent viscosities compared to high-pressure valve homogenization. This discrepancy can be attributed to the design of the homogenization chambers, as microfluidization generates a narrow distribution of shear forces, while high-pressure valve homogenization yields a much broader distribution. In contrast to chia monolayer nanoemulsions, the nanoemulsions stabilized by modified sunflower lecithin-Ch demonstrated a noteworthy improvement in their overall stability. This enhancement can be ascribed to their increased apparent viscosity and the highly charged interfaces of the droplets. Furthermore, throughout the entire refrigerated storage period, the omega-3 content in all nanoemulsions remained unchanged. Conclusions: In this study, mono and bilayer chia oil nanoemulsions were successfully obtained using modified sunflower lecithin and high-energy techniques. Microfluidization outperformed high-pressure valve homogenization, resulting in smaller droplets and increased viscosity. These findings are relevant for designing stable

Synergistic stabilization of garlic essential oil nanoemulsions by carboxymethyl chitosan/Tween 80 and application for coating preservation of chilled fresh pork

Garlic essential oil (GEO) is a potential natural antioxidant and antimicrobial agent for food preservation, but its intrinsic low water-solubility, high volatility and poor stability severely limit its application and promotion. In this work, we investigated the synergistic stabilization of the GEO-in-water nanoemulsion using carboxymethyl chitosan (CCS) and Tween 80 (TW 80). Additionally, the nanoemulsion was fabricated through high-pressure microfluidization and utilized for the coating-mediated preservation of chilled pork. The garlic essential oil nanoemulsion (GEON) with 3.0 % CCS and 3.0 % TW 80 exhibited more homogeneous droplet size (around 150 nm) and narrower size distribution, while maintained long-term stability with no significant change in size during 30 d storage. Compared with free GEO, the GEONs exhibited a higher scavenging capacity to DPPH and ABTS free radicals as well as higher inhibitory effects against Escherichia coli and Staphylococcus aureus, suggesting that the encapsulation of GEO in nanoemulsion considerably improved its antioxidant and antibacterial activities. Furthermore, the results of coating preservation experiments showed that the GEON coating effectively expanded the shelf-life of chilled fresh pork for approximately one week. Altogether, this study would guide the development of GEO-loaded nanoemulsions, and promote GEON as a promising alternative for coating preservation of chilled fresh meat.

Effects of dynamic high pressure microfluidization on the physical and chemical properties of carrot (Daucus carota L.)

Effects of dynamic high-pressure micro-fluidization (DHPM) on the physical and chemical properties of carrot juice was investigated in the present study. Carotene content, turbidity, centrifugal sedimentation rate, soluble solids content, color, and particle size and distribution of the samples homogenized with different pressures were evaluated. When compared with the control sample, carotene content, turbidity, suspension stability, centrifugal sedimentation rate, color, and particle size and distribution of the carrot juice showed a significant difference at different pressures. The result indicated that DHPM could improve the physical and chemical properties of carrot juice, such as increased carotene content, stability and affecting color positively resulting in a desirable high-quality juice for the consumer. Bangladesh J. Bot. 53(1): 123-129, 2024 (March)

High-pressure microfluidisation positively impacts structural properties and improves functional characteristics of almond proteins obtained from almond meal

Almond protein isolate (API) obtained from almond meal was processed using dynamic high-pressure microfluidisation (0, 40, 80, 120, and 160 MPa pressure; single pass). Microfluidisation caused significant reductions in the particle size and increased absolute zeta potential. SDS-PAGE analysis indicated reduction in band intensity and the complete disappearance of bands beyond 80 MPa. Structural analysis (by circular dichroism, UV–Vis, and intrinsic-fluorescence spectra) of the API revealed disaggregation (up to 80 MPa) and then re-aggregation beyond 80 MPa. Significant increments in protein digestibility (1.16-fold) and the protein digestibility corrected amino acid score (PDCAAS; 1.15-fold) were observed for the API (80 MPa) than control. Furthermore, significant improvements (P < 0.05) in the functional properties were observed, viz., the antioxidant activity, protein solubility, and emulsifying properties. Overall, the results revealed that moderate microfluidisation treatment (80 MPa) is an effective and sustainable technique for enhancing physico-chemical and functional attributes of API, thus potentially enabling its functional food/nutraceuticals application.

Characterization and safety assessment of bamboo shoot shell cellulose nanofiber: Prepared by acidolysis combined with dynamic high-pressure microfluidization

The preparation of cellulose nanofiber (CNF) using traditional methods is currently facing challenges due to concerns regarding environmental pollution and safety. Herein, a novel CNF was obtained from bamboo shoot shell (BSS) by low-concentration acid and dynamic high-pressure microfluidization (DHPM) treatment. The resulting CNF was then characterized, followed by in vitro and in vivo safety assessments. Compared to insoluble dietary fiber (IDF), the diameters of HIDF (IDF after low-concentration acid hydrolysis) and CNF were significantly decreased to 167.13 nm and 70.97 nm, respectively. Meanwhile, HIDF and CNF showed a higher crystallinity index (71.32 % and 74.35 %). Structural analysis results indicated the successful removal of lignin and hemicellulose of HIDF and CNF, with CNF demonstrating improved thermostability. In vitro, a high dose of CNF (1500 μg/mL) did not show any signs of cytotoxicity on Caco-2 cells. In vivo, no death was observed in the experimental mice, and there was no significant difference between CNF (1000 mg/kg·bw) and control group in hematological index and histopathological analysis. Overall, this study presents an environmentally friendly method for preparing CNF from BSS while providing evidence regarding its safety through in vitro and in vivo assessments, laying the foundation for its potential application in food.

Enhanced Stability of Oral Vitamin C Delivery: A Novel Large-Scale Method for Liposomes Production and Encapsulation through Dynamic High-Pressure Microfluidization.

In recent years, nanocarriers have been widely used as an effective solution for oral administration of pharmaceuticals. However, there is still an urgent need to speed up their translation to clinical practice. Cost-effective and industrially scalable methodologies are still needed. Herein, the production of vitamin C-loaded liposomes for nutraceutical purposes has been investigated and optimized by adopting a High-Pressure Homogenizer. Initially, the impact of process parameters on particles size, distributions, and morphology was explored. The findings document that the pressure and cycle manipulation allow for control over liposome size and polydispersity, reaching a maximum encapsulation efficiency exceeding 80%. This significantly improves the storage stability of vitamin C, as demonstrated by monitoring its antioxidant activity. Furthermore, the in vitro simulation of gastrointestinal digestion shows that liposomes could protect the active substance from damage and control its release in the gastrointestinal fluid. Thus, the whole nanodelivery system can contribute to enhancing vitamin C bioavailability. In conclusion, the results indicate that this innovative approach to producing vitamin C liposomes holds promise for clinical translation and industrial scale-up. Indeed, by utilizing food-grade materials and straightforward equipment, it is possible to produce stable and functional liposomes suitable for health products.

High-pressure microfluidization enhanced the stability of sodium caseinate-EGCG complex-stabilized fish oil emulsion

Improving the emulsion-stabilizing effect of protein by chemical or physical modification has been paid much attention recently. Here, sodium caseinate (CS) was treated by high-pressure-microfluidization (HPM) under 0–100 MPa, and was further complexed with (-)-epigallocatechin-3-gallate (EGCG) to form an excellent emulsifier that stabilized fish oil emulsions. Results showed that HPM treatment (especially 80 MPa) significantly changed the secondary structure of CS, and 80 MPa-PCS-EGCG had the best emulsifying and antioxidant activities. In addition, after HPM treatment and EGCG bonding, CS formed a thicker interface layer on the surface of oil droplets, which could better protect the fish oil from the influence by oxygen, temperature and ion concentration. Moreover, the fish oil emulsion stabilized by PCS-EGCG complex significantly delayed the release of free fatty acids subjected to in vitro digestion. Conclusively, HPM-treated CS-EGCG complex could be a potential emulsifier to improve the stability of fish oil emulsions.

Effect of Temperature, Oil Type, and Copolymer Concentration on the Long-Term Stability of Oil-in-Water Pickering Nanoemulsions Prepared Using Diblock Copolymer Nanoparticles.

A poly(glycerol monomethacrylate) (PGMA) precursor was chain-extended with 2,2,2-trifluoroethyl methacrylate (TFEMA) via reversible addition-fragmentation chain transfer (RAFT) aqueous emulsion polymerization. Transmission electron microscopy (TEM) studies confirmed the formation of well-defined PGMA52-PTFEMA50 spherical nanoparticles, while dynamic light scattering (DLS) studies indicated a z-average diameter of 26 ± 6 nm. These sterically stabilized diblock copolymer nanoparticles were used as emulsifiers to prepare oil-in-water Pickering nanoemulsions: either n-dodecane or squalane was added to an aqueous dispersion of nanoparticles, followed by high-shear homogenization and high-pressure microfluidization. The Pickering nature of such nanoemulsion droplets was confirmed via cryo-transmission electron microscopy (cryo-TEM). The long-term stability of such Pickering nanoemulsions was evaluated by analytical centrifugation over a four-week period. The n-dodecane droplets grew in size significantly faster than squalane droplets: this is attributed to the higher aqueous solubility of the former oil, which promotes Ostwald ripening. The effect of adding various amounts of squalane to the n-dodecane droplet phase prior to emulsification was also explored. The addition of up to 40% (v/v) squalane led to more stable nanoemulsions, as judged by analytical centrifugation. The nanoparticle adsorption efficiency at the n-dodecane-water interface was assessed by gel permeation chromatography when using nanoparticle concentrations of 4.0, 7.0, or 10% w/w. Increasing the nanoparticle concentration not only produced smaller droplets but also reduced the adsorption efficiency, as confirmed by TEM studies. Furthermore, the effect of varying the nanoparticle concentration (2.5, 5.0, or 10% w/w) on the long-term stability of n-dodecane-in-water Pickering nanoemulsions was explored over a four-week period. Nanoemulsions prepared at higher nanoparticle concentrations were more unstable and exhibited a faster rate of Ostwald ripening. The nanoparticle adsorption efficiency was monitored for an aging nanoemulsion prepared at a copolymer concentration of 2.5% w/w. As the droplets ripened over time, the adsorption efficiency remained constant (∼97%). This suggests that nanoparticles desorbed from the shrinking smaller droplets and then readsorbed onto larger droplets over time. Finally, the effect of temperature on the stability of Pickering nanoemulsions was examined. Storing these Pickering nanoemulsions at elevated temperatures led to faster rates of Ostwald ripening, as expected.