Microencapsulation of oils using whey protein/gum arabic coacervates

Microencapsulating sunflower oil, lemon and orange oil flavour was investigated using complex coacervation of whey protein/gum arabic (WP/GA). At pH 3.0–4.5, WP and GA formed electrostatic complexes that could be successfully used for microencapsulation purposes. The formation of a smooth biopolymer shell around the oil droplets was achieved at a specific pH (close to 4.0) and the payload of oil (i.e. amount of oil in the capsule) was higher than 80%. Small droplets were easier to encapsulate within a coacervate matrix than large ones, which were present in a typical shell/core structure. The stability of the emulsion made of oil droplets covered with coacervates was strongly pH-dependent. At pH 4.0, the creaming rate of the emulsion was much higher than at other pH values. This phenomenon was investigated by carrying out zeta potential measurements on the mixtures. It seemed that, at this specific pH, the zeta potential was close to zero, highlighting the presence of neutral coacervate at the oil/water interface. The influence of pH on the capsule formation was in accordance with previous results on coacervation of whey proteins and gum arabic, i.e. WP/GA coacervates were formed in the same pH window with and without oil and the pH where the encapsulation seemed to be optimum corresponded to the pH at which the coacervate was the most viscous. Finally, to illustrate the applicability of these new coacervates, the release of flavoured capsules incorporated within Gouda cheese showed that large capsules gave stronger release and the covalently cross-linked capsules showed the lowest release, probably because of a tough dense biopolymer wall which was difficult to break by chewing.     

Preparation of microcapsules with narrow-size distribution by complex coacervation: effect of sodium dodecyl sulphate concentration and agitation rate

In this paper, microcapsules with narrow-size distribution, in which the core materials are a kind of suspension containing pigment scarlet powders dispersed in dyed tetrachloroethylene with Span-80 as an emulsifier, are prepared by complex coacervation through controlling sodium dodecyl sulphate (SDS) concentration and agitation rate. The microcapsules, formed in optimized process of 0.01 wt% SDS and 800 rpm, are approximately 40 microm in diameter. The phase diagram for the gelatin/SDS/water system indicates that the concentration of SDS in the experiments is outside of the complex formation zone to form a gelatin-SDS complex. Consequently, SDS preferential adsorbs and enriches on the surface of the core droplets due to its higher surface activity. Then, gelatin deposits with SDS at the core droplet/water interface to form a primary layer of complexation. Subsequently, with the pH lower than the isoelectric point of gelatin, complex coacervate of gelatin and gum arabic grows on the primary layer surface and finally deposits on the droplets to form a secondary layer. On the whole, the research indicates that the existence of SDS not only decreases the droplet diameters and centralizes the droplets size distribution, but also accelerates coacervation of gelatin and gum arabic to reach the core droplet/water interface, forming no aggregating microcapsules.     

Preparation of microcapsules with narrow-size distribution by complex coacervation: Effect of sodium dodecyl sulphate concentration and agitation rate

In this paper, microcapsules with narrow-size distribution, in which the core materials are a kind of suspension containing pigment scarlet powders dispersed in dyed tetrachloroethylene with Span-80 as an emulsifier, are prepared by complex coacervation through controlling sodium dodecyl sulphate (SDS) concentration and agitation rate. The microcapsules, formed in optimized process of 0.01 wt% SDS and 800 rpm, are ～40 μm in diameter. The phase diagram for the gelatin/SDS/water system indicates that the concentration of SDS in the experiments is outside of the complex formation zone to form a gelatin–SDS complex. Consequently, SDS preferential adsorbs and enriches on the surface of the core droplets due to its higher surface activity. Then, gelatin deposits with SDS at the core droplet/water interface to form a primary layer of complexation. Subsequently, with the pH lower than the isoelectric point of gelatin, complex coacervate of gelatin and gum arabic grows on the primary layer surface and finally deposits on the droplets to form a secondary layer. On the whole, the research indicates that the existence of SDS not only decreases the droplet diameters and centralizes the droplets size distribution, but also accelerates coacervation of gelatin and gum arabic to reach the core droplet/water interface, forming no aggregating microcapsules.     

Role of pH in the coacervation of the systems: gelatin-water-ethanol and gelatin-water-sodium sulphate

Phase boundary determination, coacervate volume measurements and analysis of the phases have been made to assess the influence of pH on the coacervation of gelatin solutions by ethanol and sodium sulphate. Coacervation was found to be pH dependent. In the ethanol system coacervation was noticeable only within a pH range in the vicinity of the isoionic point; at other pH values either a viscous gel phase or floccules occurred. In the sodium sulphate system, coacervation occurred at all pH values examined. The effect of pH in changing the overall charge on the gelatin molecule is explained in relation to the formation of gelatin coacervates. Finally, the role of the coacervate phase in the microencapsulation of oil and solid particulates is discussed.     

Study of complex coacervation of gelatin with sodium carboxymethyl guar gum: microencapsulation of clove oil and sulphamethoxazole

Complex coacervation of gelatin with sodium carboxymethyl guar gum was studied. The coacervation was studied as a function of pH, colloid composition and concentration. The efficiency of coacervation was followed by measuring viscosity, coacervate yield and turbidity of test solutions. All the measurements suggest that, as the amount of sodium carboxymethyl guar gum (CMGG) in the colloids increases, the pH at which maximum coacervation happens decreases. Effective coacervation could be realized over the pH range of 2.5-4.0 using different compositions. The efficiency of the CMGG/gelatin system to encapsulate oils and solid particles is demonstrated by successful encapsulation of oil of cloves and sulphmethoxazole.     

Characterization of sodium carboxymethylcellulose-gelatin complex coacervation by chemical analysis of the coacervate and equilibrium fluid phases

The complex coacervation of sodium carboxymethylcellulose (SCMC) and gelatin has been characterized by chemical analyses of the coacervate and equilibrium fluid phases. The phenol-sulphuric acid (for SCMC) and Lowry (for gelatin) assays were used. Chemically analysed coacervate yield was used to predict optimum coacervation conditions, which occurred at a SCMC-gelatin mixing ratio of 3:7 at pH 3.5. The effects of pH, colloid mixing ratio and total colloid concentration on coacervate yield and composition were studied. The colloid mixing ratio, at which the peak coacervate yields occurred varied with coacervation pH. Increase in the total colloid concentration suppressed coacervation, resulting in a coacervate of higher water content. A similar coacervation mechanism was seen for two viscosity grades SCMC. However, because of the different degree of substitution of these two grades the SCMC-gelatin coacervates had different SCMC contents.     

Characterization of microcapsules by confocal laser scanning microscopy: structure, capsule wall composition and encapsulation rate.

The potential of confocal laser scanning microscope (CLSM) has been evaluated for characterizing microcapsules. The aim was to visualize the polymer distribution within the particle wall, and to localize and to quantify the encapsulated oil phase. Microcapsules were prepared by complex coacervation: the oil phase, gelatine, and arabic gum were labelled with fluorescent markers. For all compounds it was proved that fluorescence labelling did not alter physico-chemical properties critical to the encapsulation process. Labelling of the inner oil phase allowed us to identify and to localize, three-dimensionally, the encapsulated compound. A homogeneous distribution for both gelatine and arabic gum throughout the capsule wall was observed. The addition of fluorescently labelled casein as a macromolecular model compound to the coacervate resulted in an inhomogeneous distribution of casein within the wall material, the highest concentration of casein was found at the oil-wall interface. To determine the encapsulation rate, CLSM pictures of the microcapsule samples were acquired using different fluorescence labels for the microcapsule wall polymers and the incorporated oil phase, respectively. By applying computational image analysis, the volumes of the different phases were calculated. Comparing the results of non-destructive image analysis with those obtained by degradation, extraction and chemical analysis, a linear relation was found with correlation coefficients better than 0.980.     

Visualization and quantification of polymer distribution in microcapsules by confocal laser scanning microscopy (CLSM)

Confocal laser scanning microscopy (CLSM) was employed in order to characterize microcapsules. Microcapsules were prepared by complex coacervation: gelatin and arabic gum were labelled with fluorescent markers. In the capsule wall a homogeneous distribution for both gelatin and arabic gum throughout the capsule wall was depicted. By the use of CLSM and a computational image analysis the quantification of the labelled polymer in the wall material was possible. Adding fluorescently labelled casein as a macromolecular model compound to the coacervation process, a gradiental distribution in the wall material was observed with highest concentration of casein at the oil-wall interface. The influence of casein concentration on its deposition behaviour in the capsule wall was imaged successfully and thereafter quantified by computational image analysis.     

Preparation of microcapsules from complex coacervation of Gantrez-gelatin. I. Development of the technique

Trials to induce complex coacervation between two grades Gantrez-AN polymer (G-AN), and Type A gelatin were made. Physical parameters influencing the coacervation process were studied. Maximum coacervation was attained when the pH of the gelatin solution was at 6.8. Increasing the molecular weight of Gantrez decreased the ratio of combination of both polymers. The ratio for optimum coacervation was 1:4 for Gantrez-AN 119-gelatin system and 2:3 for Gantrez-AN 149-gelatin system with total colloid concentration of 2.5 g per cent w/v in both cases. High stirring speed gave almost spherical uniform coacervates. Recovery of the product as water-insoluble discrete units required the use of formaldehyde and isopropanol for coacervate denaturation and flocculation, respectively.     

Influence of pH adjustment on microcapsules obtained from complex coacervation of gelatin and acacia

Abstract

The coacervation behaviour of commercial grade gelatin and acacia mixtures was studied with five different acids to adjust the coacervation pH, i.e. HCI, HNO₃, H₂SO₄, acetic acid, and citric acid. The electrical equivalence pH value (EEP) of the polymer mixture was determined by means of a streaming current detector (SCD). With all acids-except H₂SO₄-maximum coacervate yield was observed at the EEP. Using H₂SO₄ the EEP was found at a lower pH value than compared with the point of maximum coacervate yield. The quantity of coacervate at the EEP was significantly reduced in the presence of H₂SO₄ whereas with all other acids, almost no differences were found. The dependence of the coacervate volume on the added amount of acid did not change in parallel to the dry coacervate yield and there was no coincidence of the maximum coacervate volume and the EEP. The barrier properties of the capsule shells of corresponding microcapsules using indomethacin as a model drug were examined by dissolution studies. Indomethacin microcapsules showed the slowest release rate when the coacervation pH was adjusted to the EEP and not to the pH of maximum coacervate yield. As expected from the coacervation behaviour, dissolution profiles of the microcapsules were quite similar even when different acids were used for pH adjustment.     

Spontaneous Formation of Small Sized Albumin/acacia Coacervate Particles

Abstract— Microgel coacervate particles form spontaneously on mixing aqueous solutions of oppositely charged albumin and acacia, under specific conditions of pH, ionic strength, and polyion concentration, close to but not at the optimum conditions for maximum coacervate yield. The mean particle diameter of these coacervate particles is approximately 6 μm when suspended in aqueous media, as determined by HIAC/Royco particle analysis. The geometric standard deviation of the particles falls in the range 1·2–1·9 μm. The particle size was not dependent on the method of emulsification of the coacervate in the equilibrium phase, or on the stirring speed applied during the manufacturing process. The microgel particles were stable on storage, for periods up to forty-six days, without the addition of a chemical cross-linking agent, or the application of heat. Stability was measured with respect to the change in particle size of samples stored at different temperatures. The non-cross-linked microcapsules were also shown to be stable on pH change, to pH values outside the coacervation pH range. At the optimum conditions for maximum coacervate yield the albumin/acacia system formed a very viscous coacervate phase, which was unsuitable for microcapsule preparation. The Theological properties of albumin/acacia and gelatin/acacia complex coacervates optimized for maximum coacervate yield were compared. The albumin/acacia coacervate was shown to be three orders of magnitude more viscous than the gelatin/acacia system.     

Characterization of albumin-alginic acid complex coacervation

Complex coacervation of albumin and alginic acid has been investigated to characterize this process, and to prepare a microencapsulation system suitable for the encapsulation of live cells, protein and polypeptide drugs. The optimum conditions of pH, ionic strength and total polyion concentration were in accordance with predictions based on the method of Burgess & Carless (1984). Albumin/alginic acid complex coacervation appears to fit the Vies-Aranyi model for complex coacervation. Coacervation was limited compared with other polypeptide/polysaccharide systems such as gelatin and acacia, with albumin/alginic acid complex precipitates rather than complex coacervates forming under certain conditions. In particular coacervation was limited to concentrations below 0.5% w/v. At concentrations between 0.35 and 0.5% w/v both complex coacervation and precipitation occurred, and at concentrations above 0.5% w/v only precipitation was detected. The albumin/alginic acid complex coacervate is very viscous and this together with the limited conditions governing the occurrence of coacervation makes this system unsuitable for the preparation of microcapsules.     

Stability and structure of protein-polysaccharide coacervates in the presence of protein aggregates

We have studied at pH 4.2 and three protein (Pr):polysaccharide (Pol) weight ratios (8:1, 2:1 and 1:1) the structure and stability of beta-lactoglobulin/acacia gum/water dispersions containing protein aggregates (BLG/AG/W) or free from aggregates (AF-BLG/AG/W). Phase diagrams were characteristic of complex coacervation. BLG/AG/W dispersions displayed a larger biphasic area than AF-BLG/AG/W dispersions, that moved towards the protein axis. It was concluded that protein aggregates affected complex coacervation both by entropic (size and molecular masses of aggregates) and enthalpic (surface properties of aggregates) effects. Laser light scattering measurements revealed that the particles diameter (d(43)) induced by demixing was controlled by protein aggregates in AF-BLG/AG/W dispersions. At 1 wt.% biopolymer concentration, particles were 15-20 times larger in AF-BLG/AG/W dispersions than in BLG/AG/W dispersions at (Pr:Pol) ratios of 2:1 or 1:1. Confocal scanning laser microscopy showed that AF-BLG/AG/W dispersions only contained spherical coacervates. BLG/AG/W dispersions contained both coacervates and aggregates coated with AG or/and BLG/AG coacervates. At a (Pr:Pol) ratio of 2:1 and 1:1, coacervates were vesicular or multivesicular. Coacervates were smaller in BLG/AG/W dispersions than in AF-BLG/AG/W dispersions. It was concluded that protein aggregates have the intrinsic ability to stabilize complex coacervates and could be used to design multifunctional delivery systems. This study showed that composite dispersions containing both protein aggregates embedded in protein-polysaccharide coacervates and free coacervates may be performed. In this respect, the design of protein aggregates with controlled size distribution and surface properties could open new possibilities both in the non-chemical control of coacervates stability and in the development of multifunctional delivery systems.     

Effect of processing parameters on the formation of spherical multinuclear microcapsules encapsulating peppermint oil by coacervation

The gelatin/gum arabic multinuclear microcapsules encapsulating peppermint oil were prepared by coacervation. The effect of various processing parameters, including the core/wall ratio, wall material concentration, pH value, as well as stirring speed on the morphology, particle size distribution, yield and loading was investigated. When the wall material concentration or the core/wall ratio increased, the morphology of multinuclear microcapsules changed from spherical to irregular and the average particle size increased, the optimal wall material concentration and the core/wall ratio were 1% and 2:1, respectively. The multinuclear spherical microcapsules with desired mean particle size can be manufactured by modulating the pH value and stirring speed. The ideal preparation conditions were pH 3.7 at 400 rpm of stirring speed. The yield of multinuclear microcapsules encapsulating peppermint oil by coacervation was approximately 90% and the processing parameters had very slight influence on the yield. When transglutaminase was used as the cross-linker instead of formaldehyde, morphology, mean particle size, yield and loading remained the same as that hardening with formaldehyde, but the particle size distribution became narrower.     

Effect of processing parameters on the formation of spherical multinuclear microcapsules encapsulating peppermint oil by coacervation

The gelatin/gum arabic multinuclear microcapsules encapsulating peppermint oil were prepared by coacervation. The effect of various processing parameters, including the core/wall ratio, wall material concentration, pH value, as well as stirring speed on the morphology, particle size distribution, yield and loading was investigated. When the wall material concentration or the core/wall ratio increased, the morphology of multinuclear microcapsules changed from spherical to irregular and the average particle size increased, the optimal wall material concentration and the core/wall ratio were 1% and 2:1, respectively. The multinuclear spherical microcapsules with desired mean particle size can be manufactured by modulating the pH value and stirring speed. The ideal preparation conditions were pH 3.7 at 400?rpm of stirring speed. The yield of multinuclear microcapsules encapsulating peppermint oil by coacervation was ～90% and the processing parameters had very slight influence on the yield. When transglutaminase was used as the cross-linker instead of formaldehyde, morphology, mean particle size, yield and loading remained the same as that hardening with formaldehyde, but the particle size distribution became narrower.     

The Influence of Surface Properties on Uptake of Oil into Complex Coacervate Microcapsules

Abstract— A range of surfactants with different hydrophile-lipophile balance (HLB) values was selected to investigate the influence of interfacial properties on the uptake of oil droplets into complex coacervate microcapsules. The well characterized gelatin/acacia complex coacervate system was used in this study and the encapsulation of squalane, and oleic acid was investigated. The surfactants investigated were Span 85, Span 80, Span 40, egg yolk lecithin, and Tween 80. Combinations of surfactants were utilized to obtain intermediate HLB values. The percentage oil encapsulated was determined gravimetrically, based on the initial concentration and the amount extracted from the microcapsules. The aqueous interfacial tension values of the oils and oil/surfactant systems were measured using the Wilhelmy plate method. The interfacial properties were correlated to the percentage oil uptake by the coacervate phase. The relative hydrophobicity/lipophilicity of the oil influenced its uptake by complex coacervate droplets. The presence of surfactant affected oil uptake, depending on the HLB value of the surfactant or surfactant mixture. Uptake of squalane by the gelatin/acacia coacervates was found to be optimized by the addition of surfactants with HLB values in the range 2·5–6. The percentage uptake of oil decreased rapidly for systems prepared containing surfactants with HLB values outside this range. No correlation was observed between oil uptake by the coacervate phase and the interfacial tension of the oil and oil/surfactant systems with double-distilled deionized water.     

Physical factors affecting microencapsulation by simple coacervation of gelatin

Gelatin has been coacervated at 60 degrees C using sodium sulphate. Interfacial tensions between coacervate and supernatant liquid, coacervate and two oils (with and without one of two drugs, clofibrate and chlormethiazole) and supernatant liquid and the oils (+/- drug) have been measured by a drop volume technique, in the presence and absence of one of three surfactants, cetrimide, sodium lauryl sulphate and hexadecyltrimethylammonium lauryl sulphate (double salt). Spreading coefficients calculated from tensions indicate that coacervate should spread readily over oil droplets in presence of double salt, less readily with cetrimide and spreading is unlikely in the presence of sodium lauryl sulphate. The sign of the charge on coacervate droplets and oil droplets was identified under different conditions and showed coacervate droplets and oil droplets have opposite charges except in the presence of sodium lauryl sulphate. Microcapsules were prepared using cetrimide or 'double salt' as emulsifier and release of drug measured. Those prepared with 'double salt' released more slowly than those prepared with cetrimide.     

Oxidation of linoleic acid encapsulated with gum arabic or maltodextrin by spray-drying

Linoleic acid was emulsified with gum arabic or maltodextrin at various weight ratios of the acid to the polysaccharide in the presence or absence of a small-molecule emulsifier. The emulsions were spray-dried to produce microcapsules. Emulsions prepared with gum arabic were smaller in droplet size and more stable than those prepared with maltodextrin, and linoleic acid in a gum arabic-based microcapsule was also most resistant to oxidation than that in a maltodextrin-based microcapsule. Although the oil droplet size in the emulsion with maltodextrin decreased and the emulsion stability was improved by addition of a small-molecule emulsifier to linoleic acid, the oxidative stability of the encapsulated linoleic acid was not significantly improved. Encapsulated linoleic acid of small droplet size oxidized more slowly than that of large droplet size.     

Oxidation of linoleic acid encapsulated with gum arabic or maltodextrin by spray-drying

Linoleic acid was emulsified with gum arabic or maltodextrin at various weight ratios of the acid to the polysaccharide in the presence or absence of a small-molecule emulsifier. The emulsions were spray-dried to produce microcapsules. Emulsions prepared with gum arabic were smaller in droplet size and more stable than those prepared with maltodextrin, and linoleic acid in a gum arabic-based microcapsule was also most resistant to oxidation than that in a maltodextrin-based microcapsule. Although the oil droplet size in the emulsion with maltodextrin decreased and the emulsion stability was improved by addition of a small-molecule emulsifier to linoleic acid, the oxidative stability of the encapsulated linoleic acid was not significantly improved. Encapsulated linoleic acid of small droplet size oxidized more slowly than that of large droplet size.     

Effect of the cross-linking agent and drying method on encapsulation efficiency of orange essential oil by complex coacervation using whey protein isolate with different polysaccharides

Orange essential oil was microencapsulated by complex coacervation with whey protein isolate (WPI): carboxymethylcellulose (CMC), WPI:sodium alginate (SA) and WPI:chitosan (CH). Effect of pH, protein:polysaccharide ratio and solid concentration on coacervation efficiency were selected for the best coacervation conditions. Tannic acid (TA), sodium tripolyphosphate, oxidised tannic acid and transglutaminase enzyme (TG) were used as cross-linking agents. Highest encapsulation efficiency (EE) for wet coacervated microcapsules ranged from 88% to 94%. Microcapsules were freeze and spray dried to evaluate their effect on its integrity. EE was higher than 80% in freeze dried coacervated microcapsules with and without cross-linking agent, but they formed a solid cake. Spray-dried samples formed a free fluid solid (10–20?µm), where the systems WPI:CMC and WPI:CH cross-linked with TA and TG, respectively showed the highest EE (47% and 50% respectively), representing 400% improvement compared to the samples without cross-linking.