Search

The purpose of this study was to evaluate the effects of dietary supplementation of Quillaja saponin (QS) on growth performance, nutrient digestibility, fecal microbiota, and fecal gas emission in growing pigs. A total of 50 crossbred growing pigs [(Yorkshire × Landrace) × Duroc] with an initial body weight of 23.83 ± 1.95 kg were randomly assigned to one of two treatments for a 56 d trial with five replicate pens per treatment and five pigs (two barrows and three gilts) per pen. Dietary treatments including control diet and control diet supplemented with 200 mg·kg−1 QS. The average daily gain was significantly increased during days 0–56, whereas the fecal ammonia emission on day 56 and fecal coliform bacteria counts on day 28 were significantly decreased in pigs fed with QS-containing diet. However, dietary supplementation of QS had no significant effects on apparent total tract digestibility. In conclusion, dietary supplementation of 200 mg·kg−1 QS had beneficial effects on growth performance, fecal microbiota, and fecal gas emission in growing pigs. Considering the carry-over effects, the adaption period should be at least 28 d when supplementing 200 mg·kg−1 QS to the diet of growing pigs for improving the growth performance.

Plant saponin extracts are currently of increasing importance for the environment and climate. Saponin extracts are used for the regeneration of soils contaminated by environmental pollutants such as polycyclic aromatic hydrocarbons. Furthermore, as feed additives they may have the potential to reduce the release of the greenhouse gas methane in ruminants, or to reduce ammonia emissions in pigs and broilers. Quillaja extracts play a major role in emission reduction. Some commercially available Quillaja extracts have been adulterated by blending with an inexpensive Madhuca saponins extract. Since a quantitation has not been performed so far, the present work aims at the development of a quantitative method for the determination of the main ingredient Mi-saponin A of Madhuca. For this purpose, a semi-preparative HPLC method for the isolation of Mi-saponin A from mahua seed cake, which is the press residue of the seed kernels of Madhuca longifolia, was developed. Using 1H NMR, 13C NMR, and high resolution mass spectrometry, the identity was unambiguously confirmed and the purity was excellent. It was used as standard for quantitation of Mi-saponin A in various currently purchased commercially available Quillaja saponin extracts. The concentration of Mi-saponin A was between 1.68 and 2.65 % (w/w), which reveals that the blending of Quillaja saponin extracts with Madhuca is still relevant. Moreover, the developed method for the quantitation of Mi-saponin A is also suitable for lower concentration. Thus, Mi-saponin A was found at a concentration of 0.10 % (w/w) in Gypsophila saponin extracts, confirming that there was a contamination with this Madhuca saponin.

This study reports the applicability of quillaja saponin (QS) as a vigorous and environmentally friendly shale swelling inhibitor. QS is a natural surfactant, which is extracted from herbal sources. The inhibition strength of this surfactant was assessed through various experiments, such as sedimentation, inhibition, filtration, particle size, Scanning Electron Microscope (SEM) images, and cutting recovery. Data obtained from these tests illustrated that QS greatly inhibits clays from swelling. The optimal concentration for QS in this intend was 10 g/L. Compatibility of this surfactant with other common additives was also investigated, which showed that it is totally compatible. Finally, the potential inhibition mechanism was assessed through thermal gravimetric analysis (TGA), zeta potential, and contact angle measurement experiments. Surface coating, and wettability alteration of clay particles to the oil-wet state was recognized as the most probable mechanism.

Many lipophilic polyphenols have low bioavailability because of their poor solubility and chemical stability within the human gut. The encapsulation of these polyphenols within digestible lipid droplets can improve their solubility and stability. However, there is currently a poor understanding of how the molecular and physicochemical properties of specific polyphenols impact these characteristics. In this study, the factors influencing the solubility and stability of different polyphenols (curcumin, resveratrol, and quercetin) under simulated gastrointestinal conditions were examined when they were delivered in the form of soybean oil-in-water nanoemulsions containing quillaja saponin-coated droplets (d32 ≈ 0.15 μm; ζ = -63 mV; pH 5). The polyphenols were loaded into the lipid droplets using a pH-driven method, which is based on the pH-dependent electrical charge, oil-water partitioning, and water-solubility of these molecules. The encapsulation efficiency of all three polyphenols was relatively high (75-87%). However, their chemical stability under gastrointestinal conditions (i.e., the % remaining after exposure to gastrointestinal conditions) differed considerably: quercetin (44%), curcumin (92%), and resveratrol (100%). This effect was mainly attributed to the lower logD value of quercetin (2.17) than those of resveratrol (3.39) and curcumin (4.12). As a result, a high fraction (>50%) of quercetin was located within the aqueous gastrointestinal fluids, where it would be more prone to chemical degradation or precipitation. The fraction of the polyphenols solubilized in the gastrointestinal fluids (bioaccessibility) followed a different trend: curcumin (57%) < quercetin (73%) < resveratrol (76%). This effect was attributed to the chemical instability and/or binding of curcumin with other molecules in the simulated intestinal conditions. These results provide useful information for designing nanoemulsion-based delivery systems to improve the efficacy of lipophilic polyphenols.

We systematically investigated the impact of oil droplet diameter (≈0.15, 1.6, and 11 μm) on the bioaccessibility of three oil-soluble vitamins (vitamin A palmitate, vitamin D, and vitamin E acetate) encapsulated within soybean oil-in-water emulsions stabilized by quillaja saponin. Lipid digestion kinetics decreased with increasing droplet size due to the reduction in oil-water interfacial area. Vitamin bioaccessibility decreased with increasing droplet size from 0.15 to 11 μm: 87 to 39% for vitamin A; 76 to 44% for vitamin D; 77 to 21% for vitamin E. Vitamin bioaccessibility also decreased as their hydrophobicity and molecular weight increased, probably because their tendency to remain inside the oil droplets and/or be poorly solubilized by the mixed micelles increased. Hydrolysis of the esterified vitamins also occurred under gastrointestinal conditions: vitamin A palmitate (∼90%) and vitamin E acetate (∼3%). Consequently, the composition and structure of emulsion-based delivery systems should be carefully designed when creating vitamin-fortified functional food products.

A 58-year-old non-atopic chemical worker complained about work-related asthma and rhinoconjunctivitis about 4 years after exposure to quillaja bark and soapnut. Bronchial hyperresponsiveness was demonstrated after withdrawal of medication for 12 hours. Skin prick tests with extracts from quillaja bark and soapnut from the workplace were positive, but ImmunoCAP was positive only with quillaja bark, probably due to the low protein content of the extract from soapnut. Sensitizations to quillaja bark and soapnut, but not to saponin were demonstrated by immunoblot. An inhalation test with a dosimeter was positive with the soapnut extract. A link between disease and exposure was documented by serial measurements of exhaled nitric oxide at and off work, despite preventive measures. A diagnosis of occupational allergy due to quillaja bark and soapnut was made. Further exposure reduction was recommended.

Vaccine adjuvants are compounds that enhance/prolong the immune response to a co-administered antigen. Saponins have been widely used as adjuvants for many years in several vaccines – especially for intracellular pathogens – including the recent and somewhat revolutionary malaria and shingles vaccines. In view of the immunoadjuvant potential of Q. brasiliensis saponins, the present study aimed to characterize the QB-80 saponin-rich fraction and a nanoadjuvant prepared with QB-80 and lipids (IMXQB-80). In addition, the performance of such adjuvants was examined in experimental inactivated vaccines against Zika virus (ZIKV). Analysis of QB-80 by DI-ESI-ToF by negative ion electrospray revealed over 29 saponins that could be assigned to known structures existing in their congener Q. saponaria, including the well-studied QS-21 and QS-7. The QB-80 saponins were a micrOTOF able to self-assembly with lipids in ISCOM-like nanoparticles with diameters of approximately 43 nm, here named IMXQB-80. Toxicity assays revealed that QB-80 saponins did present some haemolytical and cytotoxic potentials; however, these were abrogated in IMXQB-80 nanoparticles. Regarding the adjuvant activity, QB-80 and IMXQB-80 significantly enhanced serum levels of anti-Zika virus IgG and subtypes (IgG1, IgG2b, IgG2c) as well as neutralized antibodies when compared to an unadjuvanted vaccine. Furthermore, the nanoadjuvant IMXQB-80 was as effective as QB-80 in stimulating immune responses, yet requiring fourfold less saponins to induce the equivalent stimuli, and with less toxicity. These findings reveal that the saponin fraction QB-80, and particularly the IMXQB-80 nanoadjuvant, are safe and capable of potentializing immune responses when used as adjuvants in experimental ZIKV vaccines.

This study aims to determine the content and total levels of secondary metabolites of ethanol extract of moringa seed (Moringa oleifera Lam.). Extraction of moringa seed using the maceration method to obtain a filtrate. The filtrate obtained was concentrated using a rotary evaporator to obtain a concentrated extract. The extract was tested qualitatively for alkaloids, flavonoids, saponins, and tannins using a suitable reagent with the test parameters. The quantitative test was using UV-Visible spectrophotometry. Alkaloids using test parameters equivalent total alkaloid quinine, flavonoids use parameter test equivalent of the total flavonoids quercetin, saponin using test parameters from Quillaja total saponins and tannins quantitative bark using test parameters total tannin tannic acid equivalent. The result showed that moringa seed positive for alkaloids characterized by orange deposition, flavonoids are characterized by the formation of the yellow color orange, saponin their stable foam, and tannins are marked in black. Quantitative test results alkaloids of 916,87 µg/g, flavonoids of 0.255%, saponin of 6.367%, and tannin of 3,724.5 µg/g. Keywords: Moringa oleifera Lam., secondary metabolites, spectrophotometry

Yellow Fever Virus (YFV) and Chikungunya Virus (CHIV) are neglected reemerging pathogens that cause comorbidities worldwide. Since no antiviral drug is prescribed for those infections, there is a demand on researching compounds that inhibit viral replication. Saponins are amphiphilic compounds that already demonstrated in vitro activity against enveloped virus. Therefore, two purified saponin fractions from Quillaja spp. were evaluated regarding their antiviral potential against YFV and CHIKV. The cell line used in this study was VERO (African green monkey kidney cells) since it is permissive to the replication of both viruses. The antiviral activity of both saponins fractions was screened using the plaque reduction assay protocol. Although saponins did not inhibited YFV replication, they strongly inhibited CHIKV. To confirm the absence of antiviral activity of Quillaja saponins against YFV, the cytopathic effect inhibition assay was performed also. Further studies are required to determine the antiviral mechanisms involved in the CHIKV inhibition.

The cytotoxic mechanism of the saponin QS-21 and its aglycone quillaic acid (QA) was studied on human gastric cancer cells (SNU1 and KATO III). Both compounds showed in vitro cytotoxic activity with IC50 values: 7.1 μM (QS-21) and 13.6 μM (QA) on SNU1 cells; 7.4 μM (QS-21) and 67 μM (QA) on KATO III cells. QS-21 and QA induce apoptosis on SNU1 and KATO III, as demonstrated by TUNEL, Annexin-V and Caspase Assays. Additionally, we performed in silico docking studies simulating the binding of both triterpenic compounds to key proteins involved in apoptotic pathways. The binding energies (∆Gbin) thus calculated, suggest that the pro-apoptotic protein Bid might be a plausible target involved in the apoptotic effect of both triterpenic compounds. Although QA shows some antiproliferative effects on SNU1 cells cultured in vitro, our results suggest that QS-21 is a more powerful antitumor agent, which merits further investigation regarding their properties as potential therapeutic agents for gastric cancer.

The antimicrobial properties of Pelargonic acid (PA), a component of tomatoes, makes it an attractive candidate as a food additive and sanitizer. The antimicrobial efficacy of PA emulsions generated using surfactants: Tween 80, Triton X100, Sodium Dodecyl Sulfate (SDS) and Quillaja Saponin was evaluated against Salmonella serotypes Newport, Oranienburg and Typhimurium. Micelle/dropletsize, and minimal inhibitory concentrations (MIC) were determined. Surfactant type and concentration significantly influenced the antimicrobial efficacy of PA (p Salmonella Newport was the most (p S. Newport and 15.62 mM against S. Oranienburg/S. Typhimurium, respectively. Addition of PA to Trypticase Soy Broth resulted in a decreased growth rate and an increased lag phase duration. Cells exposed to PA formed elongated filaments (>5 µm). Additionally, Salmonella serotypes Typhimurium and Newport also formed floccular biofilms. PA emulsions at a concentration of 31.25 mM generated using 1% SDS and 1% Quillaja saponin resulted in >6 log CFU/ml reduction in Salmonella population. Althought all PA emulsions evalauted inhibited Salmonella, morphological changes to this antimicrobial varied substantially among the Salmonella serotypes tested.

Plant-derived protein can present antinutrients (ANs) in its composition. The ANs can interact with the protein, affecting its solubility and functional properties, such as gelation. This work evaluated the effect of three ANs, namely phytic acid (PA), tannic acid (TA), and Quillaja bark saponin (QBS), on the gelation and solubility of soy (SPI), pea (PPI), and rice protein isolate (RPI). The ANs altered the protein isolates gelation and solubility. PA decreased the solubility and gelation of the three protein isolates at pH 3.0. The TA was the AN that most decreased the solubility and gelation characteristics of SPI and PPI at both pHs analyzed. QBS increased the gelation of SPI at pH 3.0 but decreased the final gel strength of RPI at the same pH. These results show that the knowledge of the presence of ANs in the protein isolates is of fundamental relevance for the processing of vegetable proteins. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s13197-020-04458-7) contains supplementary material, which is available to authorized users.

This study aims to determine the content of secondary metabolites and the levels of total secondary metabolites in ethanol extract of Okra (Abelmoschus esculentus L. Moench) fruit with UV-Vis Spectrophotometry method. Okra (Abelmoschus esculentus L. Moench) fruit extract was prepared by maceration method with 96% ethanol. The extract was concentrated using a rotary evaporator that followed by a qualitative test for alkaloids, flavonoids, saponins, and tannins using suitable reagents with the test parameters, while for the qualitative assay using a UV-Vis Spectrophotometry. Qualitative analysis of alkaloids using test parameters of total alkaloids equivalent to quinine, flavonoid using parameter of total flavonoids equivalent to quercetin, saponin using parameters of standard saponins total from the Quillaja Bark, and tannin using test parameters of total tannins equivalent to tannic acid. Qualitative test results indicated that the ethanol extract of Okra fruit tested positive for alkaloids content wich characterized by the orange deposition, flavonoids characterized by the formation of an orange-yellow color, a stable foam for saponins and tannins marked in black. The quantitative analysis resulted alkaloids of 2228.06 mg/gram, flavonoids of 2.79 mg/gram, saponins of 10.03 mg/gram, and tannins of 1973.27 mg/gram. Keywords: Okra fruit, secondary metabolites, UV- Vis spectrophotometry, qualitative, quantitative.

Knowledge of binary emulsifiers' influence on the formation and stability of emulsion-based products is still limited. The aim of this study was to investigate the emulsifying properties of Quillaja saponin-egg lecithin mixtures at different concentration ratios (r = 5:0, 4:1, 3:2, 2:3, 1:4, and 0:5) with total emulsifier concentration set to 0.5% or 1.0% (w/w). For this, oil-in-water emulsions (10% oil, pH 7) were prepared via high-pressure homogenization. Furthermore, emulsion stability against different environmental stresses was tested. All the binary emulsifier mixtures formed submicron sized emulsions upon homogenization. The most stable emulsions among the mixed emulsifiers were obtained at low Quillaja saponin concentration at r = 1:4 that showed similar physical stability over time to emulsions stabilized by Quillaja saponins and egg lecithin alone. The data suggested that the mixtures of Quillaja saponins and egg lecithins built mixed interfacial layers that were prone to changes over time. Emulsions stabilized by the binary mixtures were in general less stable against changes in pH and ionic strength than the emulsions stabilized by the individual emulsifiers. An exception were the emulsions at r = 1:4 that showed improved stability at pH 2 over the phase separated Quillaja saponin-stabilized emulsions at the same pH. Moreover, all the emulsions were heat stable up to 90 °C. On the other hand, none of the emulsions were stable upon freeze-thawing. These results increase our understanding of technofunctionality of binary emulsifier systems. PRACTICAL APPLICATION: Food-grade and natural emulsifier mixtures composed of Quillaja saponins and egg lecithin may be used in selected emulsion-based food or personal care product applications to replace synthetic surfactants due to issues with consumer acceptance and regulatory restrictions.

The influence of quillaja saponin on the formation and stability of water-in-oil-in-water (W1/O/W2) emulsions was investigated. In particular, the ability of these emulsions to encapsulate iron (ferrous sulfate) in a form that would not promote lipid oxidation was examined. The emulsions were formed using a two-step process. First, iron sulfate was dissolved in the aqueous phase used to form 20 wt% water-in-oil (W1/O) emulsions stabilized by polyglycerol polyricinoleate (PGPR). Second, W1/O/W2 emulsions were fabricated by homogenizing the W1/O emulsions (20 wt%) with another aqueous phase (80 wt%) containing quillaja saponin (0.5, 1, 2, 3, and 4 wt%). The diameter of the oil droplets in the W1/O/W2 emulsions decreased with increasing saponin concentration. Confocal microscopy confirmed that the W1/O/W2 emulsions consisted of tiny water droplets entrapped inside oil droplets that were dispersed in an external water phase. Rheological analysis showed the W1/O/W2 emulsions were shear-thinning fluids. The iron encapsulation efficiency of the emulsions remained high (> 82%) throughout storage for 28 days at ambient temperature. Lipid oxidation of the iron-loaded W1/O/W2 emulsions could be retarded by incorporation of rice bran stearin in the oil phase. The double emulsions developed in this work have potential for encapsulation and delivery of iron in functional foods containing oxidizable lipids.

Solid lipid nanoparticles (SLN) have emerged as a potential delivery system for bioactive ingredients, as they are associated with improving the stability of incorporated bioactive components by preventing their diffusion to the particle surface. The aim of this study was to investigate the impact of carrier lipid composition and surfactant type on the physical stability and thermal behavior of SLN containing β-carotene. As carrier lipids we used cocoa butter and/or tristearin and as surfactants we used Quillaja saponin extract in a combination with high- or low-melting lecithin. All the ingredients were food-grade. The results showed that all the SLN were physically stable and had a particle diameter Quillaja saponins and low-melting lecithin as emulsifier, which showed slightly increased particle sizes during storage due to polymorphic transition. In general, this study indicated that the introduction of heterogenous lipids can improve the physical stability of the SLN due to the more random crystal structures formed. Moreover, the polymorphic stability is dependent on the surfactants applied. These results are of interest for manufacturers in food, pharmaceutical, cosmetic and personal care industries using cocoa butter or any lipid blend to enrich their products with bioactive components.

We evaluated the effects of Quillaja saponin (400 mg/kg) on growth performance, nutrient digestibility, fecal gas emissions, and meat quality in pigs when used as a supplement in a finishing pig diet. In total, 80 finishing pigs [(Yorkshire × Landrace) × Duroc] with an average initial body weight of 61.53 ± 4.41 kg were used in a 61-day study. Pigs were allotted to one of two treatments according to initial body weight with 10 replicate pens per treatment and four pigs (two barrows and two gilts) per pen. The dietary treatments consisted of (1) a basal diet (CON), or (2) a CON + 400 mg/kg Quillaja saponin (QS) diet. The growth performance and nutrient digestibility were not affected by supplementing the diet with QS. The ammonia emissions were decreased by Quillaja saponin supplementation (P

Background This study describes the design, optimization, and stress-testing of a novel phytocannabinoid nanoemulsion generated using high-pressure homogenization. $\text {QNaturale}^{\circledR }$ QNaturale Ⓡ , a plant-derived commercial emulsifier containing quillaja saponin, was used to stabilize the lipid phase droplets in water. Stress-testing was performed on this nanoemulsion in order to evaluate its chemical and colloidal stability under the influence of different environmental factors, encompassing both physical and chemical stressors. Methods Extensive optimization studies were conducted to arrive at an ideal nanoemulsion formulation. A coarse emulsion containing 16.6 wt% CBD-enriched cannabis distillate and 83.4 wt% carrier (soybean) oil dispersed in 10 wt% $\text {QNaturale}^{\circledR }$ QNaturale Ⓡ (1.5 wt% quillaja saponin) solution after 10 homogenization cycles at a pressure of 30,000 psi produced a stable nanoemulsion. This nanoemulsion was then subjected to the stress studies. Results The optimized nanoemulsion had an average droplet diameter of ca. 120 nm and average droplet surface ζ potentials of ca. -30 mV. It was imaged and characterized by a variety of protocols. It 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, and carbonation. pH values ≤2 and moderately high salt concentrations (> 100 mM), however, destabilized the nanoemulsion, eventually leading to phase separation. Cannabis potency, determined by HPLC, was detrimentally affected by any changes in the nanoemulsion phase stability. Conclusions 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.

This study focused on the evaluation of Quillaja saponin extracts with the additional quality designation DAB-which means the abbreviation of the German Pharmacopoeia (Deutsches Arzneibuch). This label suggests that Quillaja saponin extracts marked in this way are of pharmacopoeial quality and thus stand out from other Quillaja saponin extracts. The DAB ninth edition listed Quillaia saponin as a reagent. According to DAB, its quality must be checked by thin-layer chromatography (TLC), and three closely spaced zones in a defined retention factor (Rf) interval specify the saponin reagent. All the Quillaja saponin extracts obtained from different manufacturers and labeled as DAB quality complied with the TLC test. However, the analysis with high-performance liquid chromatography-quadrupole time-of-flight-mass spectrometry (HPLC-Q-ToF-MS) clearly showed additionally an intense peak pattern of Madhuca saponins in all measured samples. The TLC test for Mahua seed cake, which is the press residue from Madhuca longifolia, surprisingly showed the same three closely spaced zones in the defined Rf interval. The three zones could be identified as Mi-saponins from Madhuca after scraping and extracting them from the stationary phase of the TLC plate and subsequent measurement by HPLC-Q-ToF-MS. Therefore, the specification of the saponin reagent in DAB characterizes erroneously Madhuca saponins that are not listed as a saponin plant source for the saponin reagent.

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.

Consumer demand for ‘natural’ products has led the food industry to substitute synthetic ingredients with others of natural origin. Nanoemulsions are thermodynamically unstable systems, so an emulsifier is necessary to maintain their stability during processing and storage. The aim of this work was to study the replacement of a synthetic emulsifier (Tw80: Tween 80) with one of natural origin (QS: quillaja saponins), evaluating its effect on the physical stability of nanoemulsions under different treatments (ionic strength and temperature). Avocado oil-based nanoemulsions were prepared using different emulsifier mixtures: 5% soy lecithin (SL) and 1% Tw80, which was replaced by different QS proportions (1:0, 3:1, 1:1, 1:3, 0:1 Tw80:QS). Results showed that by increasing QS proportion particle size, polydispersity index and electronegativity of nanoemulsions also increased. All nanoemulsions remain stable after centrifugation process, since no creaming formation was observed. The ionic strength treatment had an effect on zeta potential values, but not on PS. The incorporation of quillaja saponin in nanoemulsions increases their thermal stability. Thus, the ternary emulsifier mixture (Tw80-QS-SL) can be used in the elaboration of pasteurized and sterilized emulsion-based food products.

The food, cosmetics, and personal care industries are increasingly interested in replacing synthetic colorants with natural alternatives. Anthocyanin is a renewable and sustainable source of plant-derived pigment. However, their application in commercial products is often limited because of their susceptibility to chemical degradation, which leads to color fading and/or a change in hue. In this study, we examined the possibility of protecting anthocyanin from degradation by encapsulating them within the inner water phase of a water-in-oil-in-water (W/O/W) emulsion. Polyglycerol polyricinoleate (PGPR) was used as a hydrophobic emulsifier, while quillaja saponin was used as a hydrophilic emulsifier. First, W/O emulsions containing 20% aqueous phase and 80% oil phase (PGPR in corn oil) were prepared using a microfluidizer. Second, W/O/W emulsions were prepared by blending 20% W/O emulsion with 80% aqueous phase (saponin in water) using a high-shear mixer. Changes in color, particle size, charge, and anthocyanin leakage of the emulsions were measured when the external aqueous phase was changed from pH 7 to 3. Anthocyanin encapsulation significantly slowed down pH-induced color changes, possibly because the internal aqueous phase had a different pH to the external one. Anthocyanin transfer between aqueous phases depended on pH, temperature, and initial location. Our results suggest that multiple emulsions may be useful for encapsulation and protection of natural colors.

Immunoadjuvant Quillaja spp. tree saponins stimulate both cellular and humoral responses, significantly widening vaccine target pathogen spectra. Host toxicity of specific saponins, fractions and extracts may be rather low and further reduced using lipid-based delivery systems. Saponins contain a hydrophobic central aglycone decorated with several sugar residues, posing a challenge for viable chemical synthesis. These, however, may provide simpler analogs. Saponin chemistry affords characteristic interactions with cell membranes, which are essential for its mechanism of action. Natural sources include Quillaja saponaria barks and, more recently, Quillaja brasiliensis leaves. Sustainable large-scale supply can use young plants grown in clonal gardens and elicitation treatments. Quillaja genomic studies will most likely buttress future synthetic biology-based saponin production efforts.

Introduction Quillaja brasiliensis (St. A. -Hil. & Tul) Mart (Quillajaceae) is a species native to South America, which is rich in saponins. Saponins are used in different industries, so there is a constant demand for this type of compound. Based on the wide range of applications for the saponins found in this species, notably as immunoadjuvants, we conducted a comprehensive study of this tree and its saponins. Objective The purpose of this work is to complete the characterisation of the immunoadjuvant saponin fraction from Q. brasiliensis leaves and further study the saponin fraction obtained from Q. brasiliensis bark. Methodology Saponin fractions were studied using mass spectrometry in combination with classical methods of monosaccharide and methylation analysis. We performed direct infusion and liquid chromatography/electrospray ionisation ion trap multiple-stage mass spectrometry (DI-ESI-IT-MSn and LC-ESI-IT-MS2 ). Results Seventy-five saponins, 21 from leaves and 54 from bark, were tentatively identified according to their molecular mass, fragmentation pattern and chromatographic behaviour. This work represents the first investigation of saponins from the bark of Q. brasiliensis and some of them presented new structural motifs not previously reported in the genus Quillaja. Conclusion The efficiency and selectivity of the data dependent LC-MS2 method allowed the rapid profiling of saponins from Q. brasiliensis. The results of the monosaccharide and methylation analysis performed in saponins from Q. brasiliensis fractions and Q. saponaria Molina (Quillajaceae) fraction gives further support to the structures proposed according to the mass spectral data, validating the strategy used in the present work.

Emulsifiers can often be used in combination to improve their functional performance. In this study, we examined the potential of combining two natural emulsifiers, hydrolyzed rice glutelin (HRG) and quillaja saponin (QS), on the formation and stability of oil-in-water emulsions. In particular, we investigated the impact of total emulsifier level, QS-to-HRG ratio, pH, ionic strength, and temperature on the mean particle diameter (d3,2), ξ-potential, creaming stability, and microstructure of the emulsions. The size of the oil droplets produced by homogenization decreased as the QS-to-HRG ratio increased from 1:4 to 1:1, but remained constant after that. The ξ-potential and dimensions of the droplets coated by the mixed emulsifiers were reasonably similar to those coated by QS alone, suggesting that the adsorbed saponins dominated the interfacial properties in the mixed systems. We also determined the physical stability of emulsions prepared using HRG, QS, or 1:1 QS-HRG after exposure to pH changes (pH 3–7), salt addition (0–500 mM NaCl, pH 7) and thermal treatment (30–90 °C 30 min, 200 mM NaCl, pH 7). QS-emulsions were stable to aggregation and creaming over a wider pH range than the other emulsions. Conversely, the mixed emulsions had better stability to high salt levels and temperatures (pH 7), which was attributed to a thicker interfacial layer causing stronger steric repulsion. This research may increase the utilization of plant-based emulsifiers in food and beverage applications.

Consumer concerns about synthetically derived food additives have increased current research efforts to find naturally occurring alternatives. This review focuses on a group of natural surfactants, the Quillaja saponins, that can be extracted from the Quillaja saponaria Molina tree. Quillaja saponins are triterpenoid saponins comprising a hydrophobic quillaic acid backbone and hydrophilic sugar moieties. Commercially available Quillaja saponin products and their composition and properties are described, and the technofunctionality of Quillaja saponins in a variety of food, cosmetic, and pharmaceutical product applications is discussed. These applications make use of the biological and interfacial activities of Quillaja saponins and their ability to form and stabilize colloidal structures such as emulsions, foams, crystallized lipid particles, heteroaggregates, and micelles. Further emphasis is given to the complexation and functional properties of Quillaja saponins with other cosurfactants to create mixed surfactant systems, an approach that has the potential to facilitate new interfacial structures and novel functionalities.

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.

Subunit vaccines consisting of highly purified antigens require the presence of adjuvants to create effective and long-lasting protective immunity. Advances on adjuvant research include designing combination adjuvants which incorporate two or more adjuvants to enhance vaccine efficacy. Previously, an oil-in-water emulsion adjuvant (OW-14) composed of mineral oil and an inexpensive gum Arabic emulsifier has been reported demonstrating enhanced and robust immune responses when used as an adjuvant in swine subunit vaccines. This study presents a modified version of OW-14 prepared with food-grade Quillaja saponin extract (OWq). In new OWq emulsion, saponin extract served as an emulsifier for stabilization of emulsion droplets and as an immunoactive compound. The use of saponins allowed to reduce the required amount of emulsifier in the original OW-14. However, emulsion stabilized with saponins demonstrated extended physical stability even at elevated temperature (37°C). The two-dose vaccination with a classical swine fever virus (CSFV) glycoprotein E2-based vaccine formulated with OWq produced higher levels of E2-specific IgG and virus neutralizing antibodies in pigs in contrast with animals that received the vaccine adjuvanted with oil only. In addition, new OWq adjuvant was safe to use in the vaccination of pigs.

A novel nanoparticle (NP) delivery system was designed for encapsulation of bioactive compounds for which quercetin was used as a model. A combination of almond gum (AG) and shellac as biopolymers using the antisolvent method was employed to fabricate the NPs. The influence of several significant preparation factors was investigated. Bare shellac particles showed extreme aggregation when exposed to pH 1.2 (gastric acid conditions), whereas the colloidal system containing 0.7% AG was capable to form NPs with particle size of 197nm with no sign of physio-chemical instability at pH 1.2 and with an encapsulation efficiency (EE) of 78%. Addition of small amounts of polysorbate 80 and Quillaja Saponin led to a drastic decrease in particle size to 90 and 116nm, respectively, with an encapsulation efficiency higher than 98%. The morphology characterization of the NPs with Cryo-SEM revealed that most NPs were spherical with rough surfaces, indicating that AG adsorbed onto the shellac surface as was also reflected by electrokineic measurements. DSC and FTIR analyses indicated that inclusion of quercetin within the NPs coincided with a transition from the crystalline to the amorphous state. To conclude, this study presented the formulation of promising nanocarriers applicable not only in the food but also in the pharmaceutical industry.

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.

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.

Saponins are surface active glycosides which can be extracted from a range of different plant species. This class of biosurfactants is of much current interest because of their biocompatibility and biosustainability, and a range of other interesting properties. The Quillaja saponin, the focus of this paper, is extensively used in foods, beverages and cosmetics due to their accessibility and agency approval. Aspects of their wider application rests with their use in combination with different synthetic surfactants, to optimise and tailor performance and functionality. Characterising and understanding the self-assembly of such mixtures is an essential requirement for that wider exploitation. The surface activity of a range of different saponins has been extensively reported, but the self-assembly properties of the Quillaja saponin, and its mixture with different synthetic surfactants has not been reported in any detail. Of particular interest is how the bidesmodic structure of the Quillaja saponin and the mixing with the different cosurfactants affects the evolution in the micelle structure, and this is the focus of this study. Small angle neutron scattering, SANS, has been used to characterise the micelle formation of the Quillaja saponin and of its mixtures with the anionic surfactant sodium dodecyl sulfate, SDS, and the nonionic surfactants, dodecaethylene glycol monododecyl ether, C12E12, and pentaethylene glycol monododecyl ether, C12E5. The Quillaja saponins form relatively small globular micelles with a relatively small aggregation number. In mixtures with SDS and the nonionic surfactants the micelles retain their globular structure. In the mixtures the micelle aggregation number evolves towards the value for the pure synthetic surfactant component, as the packing constraints change with the decreasing mole fraction of saponin in the micelle. The evolution in the micelle structure and size reflects the change in the structure and geometry of the associated surfactant components, and is largely dominated by the Quillaja structure. The results provide an interesting insight into the factors controlling the self-assembly in saponin – surfactant mixtures, and how the saponin structure affects the self-assembly.

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.