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Hypothesis This study compared the interfacial and emulsification properties of tea saponins, quillaja saponins, and Tween 80. We hypothesized that tea saponins are an effective and sustainable source of plant-based emulsifiers that could replace synthetic or animal-based emulsifiers in many commercial applications. Experiments Interfacial tension measurements were used to characterize the behavior of the three surfactants at an oil-water interface. The emulsifying properties of the surfactants were determined by preparing oil-in-water emulsions containing 10 wt% medium chain triglycerides (MCT) and varying surfactant levels (0.1–2 wt%) using high-pressure homogenization (pH 7). The impact of surfactant type on emulsion formation and stability was determined by measuring particle size, zeta–potential, microstructure, and creaming stability. Findings The tea saponins were capable of producing nano-scale droplets (d32

The removal of heavy metals from contaminated soil/water by biosurfactants depends on the ability of biosurfactant to form complexes with metals. The interaction of quillaja saponin, a nonionic natural biosurfactant, with heavy metal ions, viz. chromium(VI) and lead(II), has been studied spectrophotometrically and conductometrically in order to obtain detailed information for the saponin–metal complexes. Both the methods indicate complex formation between saponin and metal ions. Critical micelle concentration of saponin in the absence and presence of metal ions obtained by both the methods was in good agreement. The thermodynamic parameters, viz. free energy, enthalpy and entropy change, for micellization have been determined.

The influence of plant-based (gum arabic and quillaja saponin) and animal-based (whey protein isolate, WPI) emulsifiers on the production and stability of vitamin E-fortified emulsions was investigated. Their impact on lipid digestibility and vitamin bioaccessibility was also studied utilizing an in vitro gastrointestinal tract. WPI and saponin produced smaller emulsions than gum arabic. All emulsions had good storage stability at room temperature (4 weeks, pH 7). Saponin- and gum arabic-emulsions were resistant to droplet aggregation from pH 2 to 8 because these emulsifiers generated strong electrosteric repulsion. WPI-coated droplets flocculated around pH 5 due to a reduction in charge near their isoelectric point. Lipid digestion was slower in saponin-emulsions, presumably because the high surface activity of saponins inhibited their removal by bile acids and lipase. Vitamin bioaccessibility was higher in WPI- than in saponin- or gum arabic-emulsions. This information may facilitate the design of more efficacious vitamin-fortified delivery systems.

The stabilization of interfaces with mixed emulsifiers composed of natural ingredients is of increasing interest in order to modulate stability of food colloids. To this purpose, we first investigated the emulsifying properties of binary emulsifier mixtures composed of natural and food-grade ingredients. We prepared oil-in-water emulsions (10% oil) stabilized by Quillaja saponin-sodium caseinate mixtures at different concentration ratios (r = 5:0, 4:1, 3:2, 2:3, 1:4, 0:5) with the total concentration set to 0.5% and pH set to 7 or 3. Second, we tested the emulsion stability against environmental stresses. The emulsions produced with binary emulsifier mixtures at low Quillaja saponin ratios (r = 2:3 and 1:4, pH 7) showed the best overall performance in terms of generating small and physically stable emulsion droplets similarly to the emulsions stabilized with Na-caseinate or Quillaja saponins alone, whereas at higher Quillaja saponin ratios (r = 4:1, 3:2) the emulsions were unstable. At r = 2:3 and 1:4, the emulsions were also stable at ≤90 °C and ≤200 mM NaCl. Furthermore, they were also stable upon pH change to 2, whereas emulsions stabilized with Quillaja saponins alone (r = 5:0) flocculated. These results demonstrated that the emulsifying properties of mixed binary emulsifiers comprising natural components and subsequent stability of the emulsions is highly dependent on the applied concentration ratio. This study provides valuable insights into the techno-functionality of binary emulsifier systems.

Saponins form a group of plant-produced glycosides with potential as biopesticide ingredients. The environmental fate of saponins has never been fully investigated. In the present study, we use QS-18, a specific saponin from Quillaja saponaria as an example, to quantify hydrolysis under different conditions of pH, temperature and water chemical composition. Saponin hydrolysis in buffer solutions was base-catalyzed and followed first-order kinetics. Thus, hydrolysis was slow at pH 5.1 with a half-life of 330 ± 220 d (26 °C), which increases to 0.06 ± 0.01 d at pH 10.0. Hydrolysis rates were highly sensitive to temperature with an activation energy of 56.9 ± 14.2 kJ mol-1 at pH 7.2. In strong contrast, hydrolysis in lake waters (pH 6.4-8.2) produced different patterns with a fast initial dissipation of 25 to 60% of the added saponin within the first five hours, followed by an extremely slow reaction with 25 to 75% unreacted saponin left after reaction times longer than 120 h. The fast dissipation followed by slow hydrolysis in lake water was hypothesized to be attributed to sorption and/or flocculation of saponins by inorganic nanoparticles and/or solutes in the lake water followed by inactivation of hydrolysis due to the sorption/flocculation. The present study demonstrates that saponins may hydrolyze slowly under acidic and cold conditions. In addition, it demonstrates that dissipation kinetics in natural waters may deviate substantially from the kinetics predicted based on laboratory experiments with "clean" buffered solutions. This emphasizes the need for a deeper understanding of the processes affecting the dissipation kinetics of potential toxins under natural conditions, as fate models based on laboratory derived kinetic data may be seriously flawed.

Objective: The aim of this study was to assess cytotoxic activity of extracts and fractions from the Paramignya trimera root (PTR) and Phyllanthus amarus (PA) against two pancreatic cancer cell lines (primary: BxPc3 and secondary: CFPAC1). Materials and Methods: The root of PT and whole plant of PA were used in this study. The extracts and fractions from the PTR and PA were prepared using microwave-assisted extraction and high-performance liquid chromatography, respectively. The cytotoxic activity was assessed using the Dojindo Cell Counting Kit-8 assay. Results: The findings showed impressive cytotoxic capacity of the PTR extract against both pancreatic cancer cells of BxPc3 and CFPAC1 in a range of concentrations from 50 to 200 μg/mL, which was higher than those of ostruthin (67 μM), gemcitabine (50 nM), and four its fractions (50 μg/mL), and to be comparable to a saponin-enriched extract from Quillaja bark at 200 μg/mL. In contrast, the cytotoxic capacity of the PA extract and nine its fractions against these pancreatic cancer cell lines was significantly lower (P < 0.05) than those of gemcitabine (50 nM) and Quillaja bark extract (200 μg/mL) and being comparable to phyllanthin (4.8 μM). The IC50 values of the PTR extract against BxPc3 and CFPAC1 cancer cells were 32.12 and 36.65 μg/mL, respectively, which was much lower than that of the PA extract against CFPAC1 cancer cells (128.81 μg/mL). Conclusion: The outcomes obtained from this study reveal that the PTR extract is a lead source for the potential development of novel antipancreatic cancer drugs and/or functional foods.

The concentration ratio of binary surfactant systems can alter the interfacial layer properties, and consequently affect emulsion stability. To study the effect of varying concentration ratios (r, % w/w) of interfaces containing Quillaja saponins and other naturally occurring food-grade surfactants on their emulsifying properties, we evaluated particle size, ζ-potential, and appearance of 10% oil-in-water emulsions (pH 7) stabilized by Quillaja saponin - protein or Quillaja saponin - lecithin mixtures. Quillaja saponin - Na-caseinate mixtures (r = 0.3:0.2, 0.2:0.3, and 0.1:0.4) formed small emulsion droplets (d43: 0.2 – 0.3 μm), whereas at r = 0.4:0.1, the emulsions contained micron-sized droplets (d43: 1.217 ± 0.558 μm). Emulsions formed by Quillaja saponin - pea protein mixtures flocculated at r = 0.3:2.0, 0.2:3.0, and 0.1:4.0. In contrast, Quillaja saponin - rapeseed lecithin or egg lecithin mixtures were able to generate small emulsion droplets (d43: 0.2 – 0.5 μm) at all tested concentration ratios. The formation of stable emulsions using binary mixtures of naturally occurring emulsifiers was more dependent on the concentration ratio of the proteins than lecithins, indicating that the interfacial behavior of Quillaja saponins is more sensitive to the presence of proteins. This was attributed to attractive molecular interactions yielding complexes at interfaces and/or in the continuous phase. This study shows that food-grade binary surfactant mixtures can be used to form stable emulsions with properties being tunable by altering the mixing ratios between surfactants.

International audience; Quil-A is a purified extract of saponins with strong immunoadjuvant activity. While specific molecules have been identified and tested in clinical trials, Quil-A is mostly used as a totum of the Quillaja Saponaria bark extract. Quality control of the extract stability is usually based on the monitoring of specific saponins, whereas the comparison of samples with an initial chromatogram seems more appropriate. A reference fingerprint based on comprehensive two-dimensional liquid chromatography offers a rapid detection of nonconform samples. To fulfill quality control constraints, off-line configuration using basic instrumentation was promoted. Hence, reversed-phase liquid chromatography × reversed-phase liquid chromatography and hydrophilic interaction chromatography × reversed-phase liquid chromatography methods with ultraviolet and single-quadrupole mass spectrometry detection were kinetically optimized. The reversed-phase liquid chromatography × reversed-phase liquid chromatography method used a pH switch between dimensions to maximize orthogonality. Despite diagonalization, it led to a high peak capacity of 831 in 2 h. On the other hand, the combination of hydrophilic interaction chromatography and reversed-phase liquid chromatography offered a larger orthogonality but a lower, yet satisfactory peak capacity of 673. The advantages of both methods were illustrated on degraded samples, where the reversed-phase liquid chromatography × reversed-phase liquid chromatography contour plot highlighted the loss of fatty acid chains, while the hydrophilic interaction chromatography × reversed-phase liquid chromatography method was found useful to evidence enzymatic loss of sugar moieties.

Lipophilic polyphenol compounds (LPCs) are claimed to exhibit a broad spectrum of biological activities that may improve human health and wellbeing, including antioxidant, anti-inflammatory, and anti-cancer properties. Nanoemulsion-based delivery systems have been developed to encapsulate LPCs so as to increase their food matrix compatibility, physicochemical stability, and bioavailability. LPCs vary in their structural features, including the number and position of phenolic hydroxyl, ketone, and aliphatic groups, which results in different molecular, physicochemical, and gastrointestinal properties. In this study, we examined the impact of plant-based carrier oils (coconut, sunflower, and flaxseed oils) and LPC type (curcumin, resveratrol, and quercetin) on the in vitro gastrointestinal fate of polyphenols loaded into quillaja saponin-stabilized nanoemulsions. Coconut oil contains high levels of medium-chain saturated fatty acids (MC-SFAs), sunflower oil contains high levels of long-chain monounsaturated fatty acids (LC-MUFAs), and flaxseed oil contains high levels of long-chain polyunsaturated fatty acids (LC-PUFAs). The encapsulation efficiency and gastrointestinal stability of the LPCs were slightly lower in the MC than the LC oils. Differences in the gastrointestinal stability of the three LPCs were linked to differences in their oil–water partition coefficients. Some of the LPCs inhibited lipid digestion for certain oil types. In particular, resveratrol retarded the digestion of all three oils, but it still had the highest GIT stability and bioaccessibility. This study provides valuable information about the gastrointestinal fate of LPC-loaded nanoemulsions and highlights important differences in the behavior of LPCs with different characteristics. This knowledge may facilitate the design of more effective plant-based delivery systems for bioactive lipophilic polyphenols.

The discoloration of chlorophyll (Chl) by light is an ongoing issue for green beverages in the food industry. To suppress the discoloration of Chl in aqueous solution, the effects of different emulsifiers were investigated on the discoloration of Chl under ultraviolet (UV) irradiation to determine their potential application for use as food additives. Sucrose fatty acid ester (SE), sorbitan fatty acid ester (TW), and quillaja saponin (QS) were used as emulsifiers, while Triton X-100 (TX) was used for reference. The discoloration of Chl was measured using a color difference meter. The species of Chl in solution were determined using ultraviolet-visible (UV-Vis), fluorescence, and circular dichroism (CD) spectroscopy, and the particle size of Chl in solution was determined using dynamic light scattering. The Chl aggregates were observed by the observation of increased peak areas at longer wavelengths in the UV spectra of Chl, in addition to a reduced fluorescence intensity. The CD spectra showed that the Chl aggregates were arranged in a random structure. Furthermore, the average particle size of the Chl aggregates was determined to be approximately 100 nm. SE and QS were found to significantly enhance the formation of self-aggregates due to their high hydrophilicities compared to those of TW and TX. As a result, SE and QS protect themselves from light to suppress the discoloration of Chl. The present results therefore suggest that SE and QS are suitable emulsifiers to address the problem of Chl discoloration in beverages, such as green tea and vegetable juices. PRACTICAL APPLICATION: Chlorophyll (Chl), a green pigment present in vegetables and green tea, is discolored by light. In this study, it was found that emulsifiers (sucrose fatty acid ester and quillaja saponin) suppress the discoloration of Chl. The implementation of these emulsifiers as food additives would enable green tea or green vegetable juices to maintain their colors for long periods and could contribute significantly to the beverage industry.

Nanoadjuvants that combine immunostimulatory properties and delivery systems reportedly bestow major improvements on the efficacy of recombinant, protein-based vaccines. Among these, self-assembled micellar formulations named ISCOMs (immune stimulating complexes) show a great ability to trigger powerful immunological responses against infectious pathogens. Here, a nanoadjuvant preparation, based on saponins from Quillaja brasiliensis, was evaluated together with an experimental Zika virus (ZIKV) vaccine (IQB80-zEDIII) and compared to an equivalent vaccine with alum as the standard adjuvant. The preparations were administered to mice in two doses (on days zero and 14) and immune responses were evaluated on day 28 post-priming. Serum levels of anti-Zika virus IgG, IgG1, IgG2b, IgG2c, IgG3 were significantly increased by the nanoadjuvant vaccine, compared to the mice that received the alum-adjuvanted vaccine or the unadjuvanted vaccine. In addition, a robust production of neutralizing antibodies and in vitro splenocyte proliferative responses were observed in mice immunized with IQB80-zEDIII nanoformulated vaccine. Therefore, the IQB80-zEDIII recombinant preparation seems to be a suitable candidate vaccine for ZIKV. Overall, this study identified saponin-based delivery systems as an adequate adjuvant for recombinant ZIKV vaccines and has important implications for recombinant protein-based vaccine formulations against other flaviviruses and possibly enveloped viruses.

Quillaja genus (Quillajaceae family) is endemic to South America, where is represented by two species, Quillaja saponaria and Quillaja brasiliensis. One outstanding characteristic of these forest tree species is their production of saponins, a family of amphipathic glycosides, involved in the defensive response of plants against biotic and abiotic factors. Saponins are metabolites of economic importance due to their chemical and physical properties. Basic and applied research efforts performed during the last decades, mainly on Q. saponaria, have placed these compounds as an important raw material in industrial areas, such as food and beverage, cosmetics, vaccine production, biopesticides, among others. In this review, we summarize information on saponins from Quillaja species during the last years, analyzing current developments by application areas, as well as their chemical composition and properties. We also describe the general advances in revealing saponin biosynthesis pathways, related genes and Quillaja genomes, as well as the conservation status, domestication processes, and perspectives in the context of implementing genetic improvement programs.

Quillaja brasiliensis (A. St.-Hil. & Tul.) Mart. is currently one of the only two species of the only genus in the Quillajaceae D. Don (Dicotyledoneae) family. Recently, remarkable developments have been achieved regarding its domestication, propagation, and biochemical study. The first results on the structural study of the complex mixture of saponins produced by different organs of the tree have been published. Analogously to the related species Q. saponaria Molina, its saponins, either alone or in colloidal formulations, have been proved to be very effective as adjuvants in experimental vaccines, being able to elicit an early humoral and cellular response against the co-administered antigens. In consideration of the growing knowledge on Q. brasiliensis, this species constitutes a very promising botanical source for the development of valuable products for the biotechnological industry and research.

1. Two series of studies were conducted to determine the effects of a combination of ground plant material derived from Quillaja saponaria trees and Yucca schidigera plants (QY) as sources of saponin, on performance, productivity, nutrient digestibility and ileal morphology of growing broilers. In each trial, 480 Cobb male birds were allocated equally to 24 pens at one-day-of-age according to body weight 2. The studies consisted of two identical floor pen trials in which performance and nutrient digestibility were assessed and two trials where performance and ileal morphology were determined. In each trial, 0, 250 or 500 ppm QY were included in feed given to the broilers from 1-35 or 1-42 d of age, respectively. Eight (digestibility) or 12 (morphology) randomised replicate pens were used. 3. In the digestibility trials, two birds per pen were moved to metabolism cages at d 21. Excreta was collected for a five-day period (d 21 to 25) for the determination of apparent total tract digestibility of dry and organic matter, fat and ash and nitrogen retention. For intestinal morphology, ileal segments were collected from four birds/pen on d 21 to determine villus height and crypt depth. Performance data were collected in each trial series. 4. Results showed that feeding graded levels of QY produced significant linear improvements in performance and productivity at d 35, and similar linear effects were observed for N retention and all apparent digestibility measurements. Morphology data showed that birds receiving 250 and 500 ppm QY had significantly increased villus height 5. These results indicated that QY exerted a positive influence on the intestinal tract by increasing the absorptive surface and improving nutrient digestibility. These effects were considered to be associated with the performance improvements recorded in both experiments.

In this research, we assessed the detergency properties along with chemical characteristic of the surfactant extracts from the most frequently cited detergent plants in Northern Thailand, namely, Sapindus rarak, Acacia concinna, and Litsea glutinosa. Moreover, as to provide the sustainable option for production of such valuable ingredients, plant tissue culture (PTC) as alternative method for industrial metabolite cultivation was also proposed herein. The results illustrated that detergent plant extracts showed moderate in foaming and detergency abilities compared with those of synthetic surfactant. The phytochemical analysis illustrated the positive detection of saponins in L. glutinosa plant extracts. The highest callus formation was found in L. glutinosa explant cultured with MS medium supplemented with 2.0 mg/L Indole-3-acetic acid (IAA). The callus extract was chemical elucidated using chromatography, which illustrated the presence of saponin similar to those from the crude leaf and Quillaja saponin extracts. Compact mass spectrometry confirmed that the surfactant was of the steroidal diagnostic type.

Acute hyposalinity stress in shrimp ponds is a natural occurrence during the rainy season and the provision of functional ingredients that will elicit resistance to this stress is needed. Quillaja saponin (QS), a triterpenoid saponin derived from the bark of Quillaja saponaria, is reported to trigger innate immunity in vitro and in vivo in shrimps. The present study aims to evaluate its potential as growth-promoter and its immune-enhancement effects against salinity stress. Three separate experiments were done: (a) an attractability test; (b) a 30-day feeding trial to determine optimal inclusion of QS; and (3) an acute hyposalinity stress test following feeding the experimental diets. Attractability tests showed that all the Quillaja saponin inclusions significantly attracted more shrimps than the control diet (p

This study describes the design, optimization, and stress-testing of a novel phytocannabinoid nanoemulsion generated using high-pressure homogenization. QNaturale®, a plant-derived commercial emulsifier containing quillaja saponin, was used to stabilize the lipid phase droplets in water. Optimization studies revealed that after 10 homogenization cycles at a pressure of 30,000 psi in the presence of 10 wt% QNaturale® (1.5 wt% quillaja saponin), average nanoemulsion droplet diameters were ca.120 nm and average droplet surface zeta-potentials were ca. -30 mV for a lipid phase comprising 16.6 wt% CBD-enriched cannabis extract and 83.4 wt% carrier (soybean) oil. The optimized nanoemulsion proved to be stable to droplet agglomeration and phase separation upon storage under ambient conditions for 6 weeks, as well as under a variety of physical stressors such as heat, cold, dilution, andcarbonation. pH values under 2 and moderately high salt concentrations (> 100 mM), however, destabilized 0the CDCBD nanoemulsion, eventually leading to phase separation. Cannabis potency, determined by HPLC, was detrimentally affected by any changes in the nanoemulsion phase stability. Quillaja saponin stabilized cannabidiol (CBD)-enriched nanoemulsions are stable, robust systems even at low emulsifier concentrations, and are therefore significant from both a scientific as well as a commercial perspective.

Bacillus megaterium strain O1 was isolated from a soapnut (Sapindus saponaria) surface and degrades Quillaja saponin as a sole carbon source. We report the draft genome sequence of B. megaterium O1, which has an estimated size of 5.1 Mb. Study of this isolate will provide insight into mechanisms of saponin degradation.

Enterotoxigenic E. coli (ETEC) is a leading cause of moderate-to-severe diarrhoea. ETEC colonizes the intestine through fimbrial tip adhesin colonization factors and produces heat-stable and/or heat-labile (LT) toxins, stimulating fluid and electrolyte release leading to watery diarrhoea. We reported that a vaccine containing recombinant colonization factor antigen (CfaEB) targeting fimbrial tip adhesin of the colonization factor antigen I (CFA/I) and an attenuated LT toxoid (dmLT) elicited mucosal and systemic immune responses against both targets. Additionally, the toll-like receptor 4 ligand second-generation lipid adjuvant (TLR4-SLA) induced a potent mucosal response, dependent on adjuvant formulation. However, a combination of vaccine components at their respective individual optimal doses may not achieve the optimal immune profile. We studied a subunit ETEC vaccine prototype in mice using a response surface design of experiments (DoE), consisting of 64 vaccine dose-combinations of CfaEB, dmLT and SLA in four formulations (aqueous, aluminium oxyhydroxide, squalene-in-water stable nanoemulsion [SE] or liposomes containing the saponin Quillaja saponaria-21 [LSQ]). Nine readouts focusing on antibody functionality and plasma cell response were selected to profile the immune response of parenterally administered ETEC vaccine prototype. The data were integrated in a model to identify the optimal dosage of each vaccine component and best formulation. Compared to maximal doses used in mouse models (10 µg CfaEB, 1 µg dmLT and 5 µg SLA), a reduction in the vaccine components up to 37%, 60% and 88% for CfaEB, dmLT and SLA, respectively, maintained or even maximized immune responses, with SE and LSQ the best formulations. The DoE approach can help determine the best vaccine composition with a limited number of experiments and may accelerate development of multi-antigen/component ETEC vaccines.

This study investigated the loading behavior of Quillaja saponin as a model surface-active cargo on (NP) nanoparticles prepared with various hydrophobic polymers and using different organic solvents through emulsification/solvent evaporation, and the impact of NP surface hydrophobicity upon the cytotoxic and hemolytic properties of the loaded entity. A superficial monolayered arrangement of saponins on NP was established (R2 > 0.9) for all NP, as the saponin loading values complied with the Langmuir adsorption isotherm over the entire concentration range. Next, based on the measurement of interfacial tension between formulation phases, and the subsequent use of Gibb’s adsorption isotherm, the packing density (Гexc) and loading of saponins on various nanospheres could be predicted with good correlation with the actual values (R2 > 0.95). The results demonstrated that the hydrophobicity of the polymeric matrix was the major determinant of saponin packing density on the nanospheres. Finally, the impact of NP surface properties upon saponin biological interactions was investigated, where a linear correlation was found between the NP surface hydrophobicity and their hemolytic properties (R2 ≅ 0.79), and cytotoxicity against two cancer cell lines (R2 > 0.76). The surface hydrophobicity of the polymeric NP seemingly governed the NP–cell membrane binding, which in turn determined the amount of membrane-bound saponins per unit NP surface area. As the saponins exert their cytotoxicity mainly through strong permeabilization of the cell membrane, a higher amount of NP–membrane association governed by a more hydrophobic matrix can lead to higher levels of cytotoxicity. These findings highlight the importance of a detailed characterization of NP surface properties, particularly in case of surface-active cargos, for these dictate the side effects and biological interactions of the delivery system.

This work introduces VaxiPatch, a novel vaccination system comprised of subunit glycoprotein vaccine antigens, adjuvants and dermal delivery. For this study, rHA of influenza virus B/Colorado/06/2017 was incorporated into synthetic virosomes, and adjuvant liposomes were formed with QS-21 from Saponaria quillaja, with or without the synthetic TLR4 agonist 3D - (6-acyl) PHAD. These components were concentrated and co-formulated into trehalose with dye. Dermal delivery was achieved using an economical 37-point stainless steel microneedle array, designed for automated fill/finish by microfluidic dispensers used for mass production of immunodiagnostics. Vaccine and adjuvant are deposited to form a sugar glass in a pocket on the side of each of the tips, allowing skin penetration to be performed directly by the rigid steel structure. In this study, Sprague Dawley rats (n = 6 per group) were vaccinated by VaxiPatches containing 0.3 µg of rHA, 0.5 µg QS-21 and 0.2 µg 3D - (6-acyl) PHAD and dye, resulting in antigen-specific IgG titers 100-fold higher than 4.5 µg of FluBlok (p = 0.001) delivered intramuscularly. Similarly, hemagglutination inhibition titers in these animals were 14-fold higher than FluBlok controls (p = 0.01). Non-adjuvanted VaxiPatches were also compared with rHA virosomes injected intramuscularly. Accelerated shelf life studies further suggest that formulated virosomal antigens retain activity for at least two months at 60° C. Further, co-formulation of a dye could provide a visible verification of delivery based on the temporary pattern on the skin. A room-temperature-stable vaccination kit such as VaxiPatch has the potential to increase vaccine use and compliance globally.

Electrically charged food-grade biopolymers can be used to form multilayer coatings around the lipid droplets in oil-in-water emulsions using a sequential layer-by-layer electrostatic deposition approach. In principle, this approach can be used to improve the stability and enhance the functionality of food emulsions. In this study, multilayer coatings were formed from saponins, polypeptides, and polysaccharides using medium chain triglyceride (MCT) lipid droplets as templates (pH 4.0). First, an emulsion containing negatively charged lipid droplets was created using quillaja saponin (QS) as an anionic emulsifier. Second, these anionic droplets were coated with a cationic polypeptide (poly-L-lysine, PLL) to form positively-charged droplets. Finally, these cationic droplets were coated with a negatively-charged polysaccharide, either pectin (PE) or κ-carrageenan (KC), to form anionic droplets. Overall, the 1-layer emulsions had the best resistance to salt, pH, and heat, indicating that quillaja saponins were effective emulsifiers. The 2-layer emulsions had better pH-stability than the 3-layer emulsions, which tended to strongly aggregate under acidic conditions. Conversely, the 3-layer emulsions had better salt-stability than the 2-layer emulsions, which tended to aggregate strongly even at low salt levels (50–100 mM NaCl). All the emulsions were relatively stable to heating (90 °C, 30 min). Overall, our results provide useful insights into the formulation of stable multilayer emulsions from food-grade emulsifiers and biopolymers. There appears to be little advantage to using the multilayer technology to enhance the physical stability of saponin-coated lipid droplets, but there may be advantages in terms of extending their functional properties, which will be explored in future studies.

A series of studies was carried out to determine the anticoccidial effects of a product derived from plant material sourced from Quillaja saponaria and Yucca schidigera. These plants are known to contain high concentrations of triterpenoid and steroidal saponins, substances that are known to display an array of biological effects. Battery tests involving individual Eimeria acervulina, Eimeria maxima, and Eimeria tenella infections and graded levels of a quillaja/yucca combination (QY) (0, 200, 250, and 300 ppm) were conducted. Body weight gain, coccidial lesion scores, and total oocysts per gram of feces (OPG) were used to evaluate anticoccidial effects. In addition, three floor pen trials evaluated the effects of 250 ppm QY in the control coccidial infections. The first pen trial measured the effects of 250 ppm QY, both alone and in combination with 66 ppm salinomycin (Sal), in a 2 3 2 factorial treatment arrangement. Two additional 42-day pen studies assessed the effects 250 ppm QY in birds vaccinated for coccidiosis. Data from the three battery trials indicated that at doses of 250 ppm QY or more, weight gain was improved, E. acervulina and E. tenella lesion scores were reduced, and OPG was lowered. In general, OPG was reduced by about 50% across all species by 250 and 300 ppm QY. Results of the pen study indicated that 250 ppm QY and Sal, when fed individually, reduced OPG and lesion scores and improved final performance. However, when QY and Sal were administered concurrently, further significant reductions in OPG occurred. The final performance of broilers vaccinated for coccidiosis was also improved at 250 ppm QY, as was OPG at both 21 and 28 days. Thus, at QY doses of 250 ppm or more, anticoccidial activity was evident but lacked the potency exhibited by many standard anticoccidials. When combined with either Sal or a live coccidiosis vaccine, QY improved the anticoccidial effects and performance of these anticoccidial methods.

We have synthesized a number of Quillaja saponaria Molina (QS) saponin analogues with a different C28 sugar unit, which features either 3, 4-diacetyl groups or a 3, 4-cyclic carbonate group at the reducing end fucoside to mimic the naturally occurring saponin adjuvant QS-7. Immunological evaluations of these analogues in BALB/c mice indicate that truncating the C28 oligosaccharide of the natural product to the tetrasaccharide (as in 5d (β)) could retain the adjuvant’s activity in enhancing IgG1 and IgG2a productions, albeit the activity is lower than that of QS-21. Further truncation or changing stereochemistry of glycosidic linkage between the tetrasaccharide and the triterpenoid quillaic acid (QA) core or within the tetrasaccharide eliminated the saponins’ adjuvant activity in terms of IgG production. On the other hand, increasing resemblance to QS-7 increased adjuvant activity and led to saponin 3’s similar IgG1 and IgG2a activities to QS-21’s, indicating that the unique adjuvant activities of QS saponins are determined by their specific structures.

In this study, two saponins-rich plant extracts, viz. Saponaria officinalis and Quillaja saponaria, were used as surfactants in an oil-in-water (O/W) emulsion based on hempseed oil (HSO). This study focused on a low oil phase content of 2% v/v HSO to investigate stable emulsion systems under minimum oil phase conditions. Emulsion stability was characterized by the emulsification index (EI), centrifugation tests, droplet size distribution as well as microscopic imaging. The smallest droplets recorded by dynamic light scattering (droplets size v. number), one day after the preparation of the emulsion, were around 50&ndash, 120 nm depending the on use of Saponaria and Quillaja as a surfactant and corresponding to critical micelle concentration (CMC) in the range 0&ndash, 2 g/L. The surface and interfacial tension of the emulsion components were studied as well. The effect of emulsions on environmental bacteria strains was also investigated. It was observed that emulsions with Saponaria officinalis extract exhibited slight toxic activity (the cell metabolic activity reduced to 80%), in contrast to Quillaja emulsion, which induced Pseudomonas fluorescens ATCC 17400 growth. The highest-stability samples were those with doubled CMC concentration. The presented results demonstrate a possible use of oil emulsions based on plant extract rich in saponins for the food industry, biomedical and cosmetics applications, and nanoemulsion preparations.