Amphiphilic polymers facilitated solid-phase extraction coupled with ultra-performance liquid chromatography-tandem mass spectrometry for direct extraction and analysis of zearalenone and zearalanone in corn juice samples.

In this work, amphiphilic polymers synthesized from carboxylated carbon nanotubes stabilized high internal phase emulsions are demonstrated to be capable of direct extracting zearalenone and zearalanone in samples consisting of an oil-water emulsion system. Under optimal conditions, the maximum adsorption capacities for zearalenone and zearalanone are 17.27 and 13.26 mg/g. The adsorption is mainly attributed to π-π interaction, hydrophobic interaction, and hydrogen-bonding interaction for zearalenone and zearalanone. The adsorption isotherms reveal that the adsorption of zearalenone and zearalanone on amphiphilic polymers synthesized from carboxylated carbon nanotubes stabilized high internal phase emulsions follows Freundlich model with multilayer and heterogeneous adsorption due to the presence of multiple kinds of adsorption sites. The relative recoveries of the spiked zearalenone and zearalanone in corn juice samples range from 85% to 93% with relative standard deviations lower than 3.52%. The results manifest the high efficiency of amphiphilic polymers synthesized from carboxylated carbon nanotubes stabilized high internal phase emulsions for the adsorption and separation of analytes in the oil-water emulsion system. This study provides a new perspective on adsorbent engineering for the adsorption application in heterogeneous media.

Chitosan/Virgin-Coconut-Oil-Based System Enriched with Cubosomes: A 3D Drug-Delivery Approach.

Emulsion-based systems that combine natural polymers with vegetable oils have been identified as a promising research avenue for developing structures with potential for biomedical applications. Herein, chitosan (CHT), a natural polymer, and virgin coconut oil (VCO), a resource obtained from coconut kernels, were combined to create an emulsion system. Phytantriol-based cubosomes encapsulating sodium diclofenac, an anti-inflammatory drug, were further dispersed into CHT/VCO- based emulsion. Then, the emulsions were frozen and freeze-dried to produce scaffolds. The scaffolds had a porous structure ranging from 20.4 to 73.4 µm, a high swelling ability (up to 900%) in PBS, and adequate stiffness, notably in the presence of cubosomes. Moreover, a well-sustained release of the entrapped diclofenac in the cubosomes into the CHT/VCO-based system, with an accumulated release of 45 ± 2%, was confirmed in PBS, compared to free diclofenac dispersed (80 ± 4%) into CHT/VCO-based structures. Overall, the present approach opens up new avenues for designing porous biomaterials for drug delivery through a sustainable pathway.

Phenolic Compound Profile and Antioxidant Capacity of Flax (Linum usitatissimum L.) Harvested at Different Growth Stages.

The profile of phenolic compounds changes during the growth of a plant and this change affects its antioxidant potential. The aim of this research has been to find the growth stage of flax with the highest antioxidant capacity, and to determine the phenolic compounds responsible for such a capacity. Flax was harvested in six growth stages: from stem extension to mature seeds. The phenolic compounds were identified using LC-TOF-MS and quantified in an extract and in the fresh matter (FM) of each growth stage. The radical scavenging activity against ABTS•+ and DPPH•, the ferric-reducing antioxidant power (FRAP), and the antioxidant activity in the ß-carotene-linoleic acid emulsion system were determined. Mono- and di-C-glycosyl flavones were found to be the most abundant phenolics of the aerial parts of flax, which also showed the highest content of isoorientin (210-538 µg/g FM). Coniferin, its derivative, and hydroxycinnamic acid derivatives were also detected. The plant was richer in flavone C-glycosides from stem extension to seed ripening (1105-1413 µg/g FM) than at the mature seed stage (557 µg/g FM). Most of the individual flavone C-glycoside contents in the extracts decreased when increasingly older plants were considered; however, the isoorientin content did not change significantly from the steam extension to the seed ripening stages. The antiradical activity against ABTS•+ and FRAP was higher for the aerial parts of the flax harvested at the flowering, brown capsule, and seed ripening stages, mainly due to the presence of flavone C-glycosides. The oxidation of ß-carotene-linoleic acid emulsion was instead inhibited more effectively by the extracts from plants at the brown capsule and mature seed stages. Coniferin and its derivative were significantly involved in this activity. The extracts from the aerial parts of the flax harvested from flowering to seed ripening could be a valuable source of flavone C-glycosides for use as nutraceuticals and components of functional foods.

High-efficiency bioconversion of phytosterol to bisnoralcohol by metabolically engineered Mycobacterium neoaurum in a micro-emulsion system.

21-Hydroxy-20-methylpregn-4-en-3-one (4-HBC, bisnoralcohol) is a crucial intermediate for the synthesis of steroidal drugs. Significant challenges including by-products formation and poor substrate solubility were still confronted in its main synthetic route by microbial conversion from phytosterol. Construction of a direct bioconversion pathway to 4-HBC and an efficient substrate emulsion system is therefore urgently required. In this study, three novel isoenzymes of 3-ketosteroid-Δ1-dehydrogenase (KstD) and 3-ketosteroid 9α-hydroxylase (KsH) in Mycobacterium neoaurum were excavated and identified as KstD4, KstD5, and KsHA3. A strain capable of fully directing the synthesis of 4-HBC was metabolically engineered via serial genetic deletion combined with enhanced expression of cholesterol oxidase (ChOx2) and enoyl-CoA hydratase (EchA19). Moreover, a micro-emulsion system combined with soybean oil and hydroxypropyl-ß-cyclodextrin improved substrate solubility and bioavailability. In batch fermentation, molar yield of 96.7% with 39.5 g L-1 4-HBC was obtained from 50 g L-1 phytosterol. Our findings demonstrate the potential for industrial-scale biosynthesis of 4-HBC.

Vine-Twining Inclusion Behavior of Amylose towards Hydrophobic Polyester, Poly(ß-propiolactone), in Glucan Phosphorylase-Catalyzed Enzymatic Polymerization.

This study investigates inclusion behavior of amylose towards, poly(ß-propiolactone) (PPL), that is a hydrophobic polyester, via the vine-twining process in glucan phosphorylase (GP, isolated from thermophilic bacteria, Aquifex aeolicus VF5)-catalyzed enzymatic polymerization. As a result of poor dispersibility of PPL in sodium acetate buffer, the enzymatically produced amylose by GP catalysis incompletely included PPL in the buffer media under the general vine-twining polymerization conditions. Alternatively, we employed an ethyl acetate-sodium acetate buffer emulsion system with dispersing PPL as the media for vine-twining polymerization. Accordingly, the GP (from thermophilic bacteria)-catalyzed enzymatic polymerization of an α-d-glucose 1-phosphate monomer from a maltoheptaose primer was performed at 50 °C for 48 h in the prepared emulsion to efficiently form the inclusion complex. The powder X-ray diffraction profile of the precipitated product suggested that the amylose-PPL inclusion complex was mostly produced in the above system. The 1H NMR spectrum of the product also supported the inclusion complex structure, where a calculation based on an integrated ratio of signals indicated an almost perfect inclusion of PPL in the amylosic cavity. The prevention of crystallization of PPL in the product was suggested by IR analysis, because it was surrounded by the amylosic chains due to the inclusion complex structure.

Novel Carboxymethyl Cellulose-Based Hydrogel with Core-Shell Fe3O4@SiO2 Nanoparticles for Quercetin Delivery.

A nanocomposite composed of carboxymethyl cellulose (CMC) and core-shell nanoparticles of Fe3O4@SiO2 was prepared as a pH-responsive nanocarrier for quercetin (QC) delivery. The nanoparticles were further entrapped in a water-in-oil-in-water emulsion system for a sustained release profile. The CMC/Fe3O4@SiO2/QC nanoparticles were characterized using dynamic light scattering (DLS), Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), a field emission scanning electron microscope (FE-SEM), and a vibrating sample magnetometer (VSM) to obtain insights into their size, stability, functional groups/chemical bonds, crystalline structure, morphology, and magnetic properties, respectively. The entrapment and loading efficiency were slightly improved after the incorporation of Fe3O4@SiO2 NPs within the hydrogel network. The dialysis method was applied for drug release studies. It was found that the amount of QC released increased with the decrease in pH from 7.4 to 5.4, while the sustained-release pattern was preserved. The A549 cell line was chosen to assess the anticancer activity of the CMC/Fe3O4@SiO2/QC nanoemulsion and its components for lung cancer treatment via an MTT assay. The L929 cell line was used in the MTT assay to determine the possible side effects of the nanoemulsion. Moreover, a flow cytometry test was performed to measure the level of apoptosis and necrosis. Based on the obtained results, CMC/Fe3O4@SiO2 can be regarded as a novel promising system for cancer therapy.

Study on the stability of deoxyArbutin in an anhydrous emulsion system.

The skin-whitening agent, deoxyArbutin, is a potent tyrosinase inhibitor that is safer than hydroquinone and arbutin. However, it is thermolabile in aqueous solutions, where it decomposes to hydroquinone. Pharmaceutical and cosmetic emulsions are normally oil-in-water (o/w) or water-in-oil (w/o) systems; however, emulsions can be formulated with no aqueous phase to produce an anhydrous emulsion system. An anhydrous emulsion system could offer a stable vehicle for compounds that are sensitive to hydrolysis or oxidation. Therefore, to enhance the stability of deoxyArbutin in formulations, we chose the polyol-in-silicone, anhydrous emulsion system as the basic formulation for investigation. The quantity of deoxyArbutin and the accumulation of hydroquinone in both hydrous and anhydrous emulsions at various temperatures were analyzed through an established high performance liquid chromatographic (HPLC) method. The results indicated that water increased the decomposition of deoxyArbutin in the formulations and that the polyol-in-silicone, oil-based, anhydrous emulsion system provided a relatively stable surrounding for the deoxyArbutin that delayed its degradation at 25 °C and 45 °C. Moreover, the composition of the inner hydrophilic phase, containing different amounts of glycerin and propylene glycol, affected the stability of deoxyArbutin. Thus, these results will be beneficial when using deoxyArbutin in cosmetics and medicines in the future.

Enzymatic enrichment of n-3 polyunsaturated fatty acid glycerides by selective hydrolysis.

Most natural oils are low in n-3 polyunsaturated fatty acids (n-3 PUFAs) content, which limits their application in health products. In this study, n-3 PUFAs in glyceride form were selectively enriched by lipase-mediated hydrolysis of n-3 PUFA-containing oils. First, commercial lipases were screened, and the lipase AY "Amano" 400SD from Candida cylindracea was the best choice in producing n-3 PUFA glycerides from tuna oil. Subsequently, the hydrolysis conditions were optimized. Under the optimal conditions, the highest n-3 PUFA content in the glyceride fraction was found to be 57.7% after enzymatic hydrolysis. Addition of Ca2+ to the system significantly shortened the reaction time from 10 to 4 h. When algal oil was used as substrate, total PUFA contents in the glyceride fraction were 89.9%. This study provides an efficient enzymatic process to produce n-3 PUFA-enriched glyceride concentrates and demonstrates that AY "Amano" 400SD can effectively discriminate against n-3 PUFAs during hydrolysis.

Development, Characterization, and Clinical Investigation of a New Topical Emulsion System Containing a Castanea sativa Spiny Burs Active Extract.

The study focused on the development and characterization of an O/W emulsion for skincare containing Castanea sativa spiny burs extract (CSE) as functional agent. The emulsion was stable and had suitable physicochemical and technological properties for dermal application and CSE showed no cytotoxicity in spontaneously immortalized keratinocytes (HaCaT) at active concentrations. A single-blind, placebo-controlled, monocentric study was designed to evaluate the skin tolerability and the skin performance of the CSE-loaded emulsion on healthy human volunteers. An improvement was observed in skin biomechanical properties such as hydration, skin elasticity and a reduction in the periorbital wrinkles in 30 days without altering the skin barrier function, sebum, pH, and erythema values. A significant skin moisturizing effect was detected while the skin barrier function was preserved. The selected natural ingredient combined with the designed formulation and the optimized preparation method has led to a final product that satisfies the physico-chemical and technological requirements underlying the safety of use and the formulative stability over time. With no negative skin reactions and highly significant effects on skin elasticity, wrinkles, and moisturization, the CSE-based emulsion achieved very satisfying outcomes representing a promising functional formulation for skin care.

Using a stable pre-emulsified canola oil system that includes porcine plasma protein hydrolysates and oxidized tannic acid to partially replace pork fat in frankfurters.

The objective of this study was to evaluate the effect of two different forms of canola oil (pure liquid form or a pre-emulsified form that includes porcine plasma protein hydrolysates modified by oxidized tannic acid) used for pork back-fat substitutions on the physico-chemical characteristics of frankfurters. When compared to the control group, partial replacement of pork back-fat using a pre-emulsified canola oil system exhibited excellent water and fat binding capacities, quicker relaxation times and lower amounts of saturated fatty acids (SFAs) (P < .05), as well as increased hardness, gumminess, and chewiness (P < .05) when verified by dynamic rheology analysis. Additionally, higher replacement ratio of pre-emulsified canola oil did significantly decrease the overall acceptability than the control group (P < .05). Our results indicate that pre-emulsified canola oil provided greater improvement with respect to the physical characteristics of partial pork back-fat replaced frankfurters, when compared to pure canola oil inclusion.

Anticancer and antibacterial effects of a clove bud essential oil-based nanoscale emulsion system.

BACKGROUND AND PURPOSE: The essential oil derived from clove buds (Syzygium aromaticum) has been used as a chemopreventive agent in Ayurvedic medicine. The antiviral, antibacterial, and anticancer properties of its chemo-skeleton have motivated this study to explore its efficacy in pharmaceutics. METHODS: Nanoscale-based emulsions were prepared by employing a spontaneous emulsification technique through self-assembly using varying concentrations of Tween 20 and Tween 80 surfactants. Their physicochemical properties and stability were studied in order to choose an optimum formulation which was clear and stable. The cytotoxicity of the stable oil-based emulsion system was evaluated using MTT assay, colony formation assay, and Annexin V-FITC assay against the thyroid cancer cell line (HTh-7). RESULTS: All three methods verified apoptosis and reduction in cancer cell proliferation, making the formulation a promising candidate as an alternative cancer drug. The oil-based emulsion system was also tested for its antibacterial properties against Staphylococcus aureus. Membrane permeability studies proved its efficacy to permeate through cell membrane, thereby increasing the leakage of cytoplasmic contents. CONCLUSION: Many current treatments for cancers are aggressive yet ineffective. This study positions the clove bud-based nanoscale emulsion as a suitable candidate for further in vivo studies and trials as a cancer drug.

Grafting phenolics onto milk protein via conjugated polymerization for delivery of multiple functionalities: Synthesis and characterization.

A synthetic scenario for functionalization of ß-lactoglobulin (ßLg) with polymeric units containing caffeic acid (ßLg-polyCA) was developed; and all intermediates and final products were structurally confirmed using nuclear magnetic resonance spectroscopy, matrix assisted laser desorption ionization time-of-flight mass spectrometry, and physico-chemically characterized using differential scanning calorimetry and circular dichroism. The antioxidant properties and emulsion stability of ßLg, ßLg-CA conjugate and ßLg-polyCA based systems containing high percentage of fish oil (50%) were evaluated; and ßLg-polyCA presented the highest antioxidant and free radical-scavenging activity based on DPPH, ABTS and HS scavenging assays (92.4, 87.92 and 67.35% respectively). Thiobarbituric acid (TBARS) test demonstrated that compared to native ßLg, ßLg-polyCA afford up 4-5 fold of inhibition of oxidative rancidity and displayed drastic secondary structure changes. Compared to native ßLg based emulsions, ßLg-polyCA had larger oil droplet sizes, stronger negative zeta potentials (-69.9 mv), narrower size distributions (PDI: 0.22) and less creaming index.

Recoverable amphiphilic polyoxoniobates catalyzing oxidative and hydrolytic decontamination of chemical warfare agent simulants in emulsion.

Amphiphilic polyoxoniobates (PONbs), [CnH2n+1N(CH3)3]7HNb6O19 (for 1, n=14; for 2, n=16; and for 3, n=18), were successfully prepared by the electrostatic interaction of hexaniobate anions with quaternary ammoniums containing long alkyl chain, and thoroughly characterized by using various techniques including Fourier transform infrared (FT-IR), 1H nuclear magnetic resonance (1H NMR), 13C nuclear magnetic resonance (13C NMR), thermogravimetric analysis (TGA), and elemental analysis. All three PONbs were used in the emulsion systems to catalytically decontaminate chemical warfare agent simulants and the influences of quaternary ammonium cations, polyanions, and amount of catalyst on the catalytic efficiency have been evaluated. Under optimal conditions, catalyst 3 in emulsion can completely convert both a the nerve agent simulant, diethyl cyanophosphonate (DECP), in 2h by hydrolysis and a sulfur mustard simulant, 2-chloroethyl ethyl sulfide (CEES), in 20min by oxidation using nearly stoichiometric 3% aqueous H2O2. Moreover, the amphiphilic catalyst 3 can be simply separated and readily reused for five recycles without obvious losing its activity. To the best of our knowledge, this study represents the first example where the emulsions of PONbs are used in the catalysis.

Adding functionality to milk-based protein: Preparation, and physico-chemical characterization of ß-lactoglobulin-phenolic conjugates.

Multi-functional phenolic emulsifiers were prepared by covalently coupling ß-Lactoglobulin (ßLg) to caffeic acid (CA) using crosslinker chemistry at different pH conditions (pH 2.5, 6.0, and 8.5). The resulting bioconjugates were characterized by MALDI-TOF MS, differential scanning calorimetry (DSC), fluorescence-quenching, infrared and circular dichroism spectroscopies. Furthermore, the emulsifying and antioxidant properties of ßLg-CA conjugates were evaluated and compared to native ß-Lactoglobulin and the non-covalent ß-lactoglobulin/caffeic complex (ßLg/CA). Results showed: 1) An optimal molar ratio (8:1) of caffeic acid to ßLg was obtained at pH 6; 2) DPPH activity of ßLg-CA increases as the number of CA units coupled increases; 3) ßLg-CA conjugates displayed comparable or superior water solubility than native ßLg and ßLg/CA. Moreover, DSC results showed that coupling of CA with ßLg significantly increased the thermal stability of ßLg. In summary, ßLg-CA conjugates can act as effective antioxidant emulsifiers and stabilizers and may find application in food and cosmetic industries.

Effects of Pre-Converted Nitrite from Red Beet and Ascorbic Acid on Quality Characteristics in Meat Emulsions.

We investigated the effects of fermented red beet extract and ascorbic acid on color development in meat emulsions. The pH of meat emulsions containing red beet extract decreased with an increase in the amount of extract added. The redness of the treated meat emulsions was higher than that of the control with no added nitrite or fermented red beet extract (p<0.05), though the redness of the meat emulsions treated with fermented red beet extract only was lower than in that treated with both fermented red beet extract and ascorbic acid (p<0.05). The highest VBN, TBARS, and total viable count values were observed in the control, and these values in the meat emulsions treated with fermented red beet extract were higher than in that treated with both fermented red beet extract and ascorbic acid (p<0.05). E. coli and coliform bacteria were not found in any of the meat emulsions tested. Treatment T2, containing nitrite and ascorbic acid, had the highest overall acceptability score (p<0.05); however, there was no significant difference between the T2 treatment and the T6 treatment, which contained 10% pre-converted nitrite from red beet extract and 0.05% ascorbic acid (p>0.05). The residual nitrite content of the meat emulsions treated with ascorbic acid was lower than in those treated without ascorbic acid (p<0.05). Thus, the combination of fermented red beet extract and ascorbic acid could be a viable alternative to synthetic nitrite for the stability of color development in meat emulsions.