Patent Publication Number: US-10773231-B2

Title: Method for producing colloidosome microcapsules

Description:
TECHNICAL FIELD 
     This invention relates to a process for the preparation of colloidosomes-type microcapsules, which employs nano- or microparticles of water-insoluble solids obtained by ionic gelation as an emulsifying material and microcapsule shell-forming. The particles that stabilize the emulsion are fixed at the interface by polyelectrolyte adsorption, cross-linking, heat treatment or treatment with fatty acid emulsions. 
     INVENTION BACKGROUND 
     A colloidosome is defined as a microcapsule whose shell is formed by colloidal particles (with particle sizes usually less than 1 μm), which are adsorbed on the encapsulated substance and subsequently stabilized forming a shell by its cross-linking, fusion or sintering (Rossier, 2009). 
     The first report of colloidosomes-type microcapsules elaboration corresponds to the Velev and cols (1996) work, which used latex particles to stabilize and form 1-octanol colloidosomes. The method called soft template (soft template, (Rossier-Miranda et al., 2009)), uses the emulsion droplets as the base on which the colloidal solid particles forming the shell are adsorbed generating greater control and adsorption efficiency. In the method called hard template (hard template) solid particles are used as the colloidosome template. 
     Dismore and cols (2002) made colloidosomes without pretreatment, using polymethylmethacrylate (PMMA) particles immersed in a continuous organic phase of decahydronaphthalene in the drops of water presence. This method allows controlling the temperature and the heating time to adjust the porosity and strength of the shell, achieving versatility in the release kinetics of the encapsulated compounds. However, the high vitreous transition temperature of PMMA (92-142° C.) limits its use in thermolabile systems. 
     Guillot (2009) and Fujiwara (2010) reported colloidosomes made from clay minerals (v.g. montmorilonite and Laponite) and sodium silicate in ammonium acid carbonate (NH 4 HCO 3 ) presence, using a W/O/W double emulsion as template. The proposed procedures, although successful in the colloidosome-type particles formation, are very difficult to scale at the industrial level and are more profiled for academic applications that allow the study and characterization of structured systems at microscopic level. 
     Liu and cols (2010) reported the production of hollow colloidosomes using nanosheets of Mg/Al double hydroxide in layers (LDH for its acronym in English). The interface structure allowed to be modulated depending on the type of organic solvent used for the emulsified oil phase extraction. Even though this modulation type of the interface structure of the colloidosome-type microcapsule is a great prospect for food applications, the use of organic solvents and the long times required for the adsorption processes, make non-viable its scaling at an industrial level. 
     The document WO 2009091255 discloses a process where colloidal particles of triglycerides, monoglycerides or diglycerides, proteins and cells are used as emulsion stabilizers and proteins and/or polysaccharides to fix the shell by a coacervation process. The main disadvantage of this process is the mono, di and triglycerides use, which, when melted, destabilize the colloidosome shell. 
     Wang and cols (2012) elaborated CaCO 3  colloidosomes by sunflower oil emulsions generation and subsequent fixation of the solid particles in the shell, by CaCO 3  crystals coprecipitation by reacting CaCl 2 ) with CO 2  in-situ. The colloidosomes generated with this methodology showed high resistance to the system water evaporation, as well as delaying the flavors release. While this process is feasible to be scaled at the industrial level, high concentrations of NaCl as a byproduct are generated, which must be eliminated by an additional washing step. 
     The document WO2009/037482 discloses the colloidosome-type microcapsules elaboration, using sterically stabilized polymer particles in the emulsions elaboration with the subsequent formation of the shell by increasing the system temperature at a temperature higher than the polymer vitreous transition. In this process a lower particles adsorption is generated on the interface with the consequent instability of the system when drying. 
     The patent WO2009/148598 describes the microfluidization technology use for the stabilized drops elaboration with polymeric, colloidal or lipid particles with potential use as an encapsulating medium for interest substances. The main drawback of this process is the use of polymer particles that necessarily require heat treatment for its sintering and forming a stable shell. 
     Obviously, the prior art shows the need to develop and optimize processes for obtaining colidosomes-type microcapsules from materials with application in the food industry, in the cosmetics field or in the pharmaceutical area. 
     BRIEF INVENTION DESCRIPTION 
     This invention relates to a process for colloidosomes-type microcapsules preparation. By means of an ionic gelation process on nano- and micrometric solid particles, the surface chemistry of these particles is modified to produce emulsions of O/W type. The particles adsorbed on the emulsion drops are subsequently fixed by charged macromolecules adsorption and polyvalent ions addition, by heat treatment, or by cross-linking. 
     Once the particles are fixed, the suspension can be dried to obtain powder colloidosomes. 
     By means of the invention process, microcapsules are obtained by using shell-forming particles with a high adsorption on the oil-water interface, which allow increasing the efficiency of the emulsification process and the encapsulated material concentration. 
    
    
     
       BRIEF FIGURES DESCRIPTION 
         FIG. 1 . Calcium carbonate particle size distribution superficially modified by ionic gelation: i) dry and agglomerated (red line); ii) milled and in aqueous suspension (green line) (Example 1). 
         FIG. 2 . Optical micrograph of oleic acid emulsion stabilized with calcium carbonate microparticles superficially modified by ionic gelation (Example 1). 
         FIG. 3 . Particle size distribution of oleic acid emulsion stabilized with calcium carbonate microparticles superficially modified by ionic gelation (Example 1). 
         FIG. 4 . Scanning electron microphotograph of oleic acid colloidosome with spray-dried calcium carbonate shell (Example 1). 
         FIG. 5 . Particle size distribution of oleic acid colloidosomes with spray-dried calcium carbonate shell (Example 1). 
         FIG. 6 . Colloidosome thermogram of oleic acid with spray-dried calcium carbonate shell (Example 1). 
         FIG. 7 . Particle size distribution of sub-micrometer titanium dioxide superficially modified by ion gelation: i) dry and agglomerated (red line); ii) milled and in aqueous suspension (green line) (Example 2). 
         FIG. 8 . Optical Micrograph of stabilized sunflower oil emulsion with sub-micrometric particles of titanium dioxide superficially modified by ionic gelation (Example 2). 
         FIG. 9 . Particle size distribution of stabilized sunflower oil emulsion with sub-micrometer titanium dioxide superficially modified by ionic gelation (Example 2). 
         FIG. 10 . Scanning electron micrographs of sunflower oil colloidosomes with spray-dried titanium dioxide shell (Example 2): left side increase of 500×, right side increase of 4300×. 
         FIG. 11 . Particle size distribution of sunflower oil colloidosomes with spray-dried titanium dioxide shell (Example 2). 
         FIG. 12 . Sunflower oil colloidosomes thermogram with spray-dried titanium dioxide shell (Example 2). 
         FIG. 13 . Particle size distribution of micrometer calcium phosphate superficially modified by ionic gelation: i) dry and agglomerated (red line); ii) milled and in aqueous suspension (green line) (Example 3). 
         FIG. 14 . Optical micrograph of mono and diglyceride mixture emulsion stabilized with micrometric calcium phosphate particles superficially modified by ionic gelation (Example 3). 
         FIG. 15 . Particle size distribution of mono and diglyceride mixture emulsion stabilized with micrometric calcium phosphate particles superficially modified by ionic gelation (Example 3). 
         FIG. 16 . Scanning electron micrographs of mono and diglyceride mixture colloidosomes with spray-dried calcium phosphate shell (Example 3): left side increase of 500×, right side increase of 5000×. 
         FIG. 17 . Particle size distribution of mono and diglyceride mixture colloidosome with spray-dried calcium phosphate shell (Example 3). 
         FIG. 18 . Colloidosome thermogram of mono and diglyceride mixture with spray-dried calcium phosphate shell (Example 3). 
         FIG. 19 . Particle size distribution of micrometric kaolin superficially modified by ionic gelation: i) dry and agglomerated (red line); ii) milled and in aqueous suspension (green line) (Example 4). 
         FIG. 20 . Optical micrograph of canola oil emulsion stabilized with micrometric kaolin particles superficially modified by ionic gelation (Example 4). 
         FIG. 21 . Particle size distribution of canola oil emulsion stabilized with micrometric kaolin particles superficially modified by ionic gelation (Example 4). 
         FIG. 22 . Scanning electron micrograph of canola oil colloidosomes with spray-dried kaolin shell (Example 4). 
         FIG. 23 . Optical micrograph of stabilized canola oil colloidosome with kaolin micrometric particles superficially modified by ionic gelation and with fatty acid reinforcement (Example 4). 
         FIG. 24 . Scanning electron micrograph of canola oil colloidosomes with kaolin shell, reinforced with fatty acids and spray-dried (Example 4). 
         FIG. 25 . Canola oil colloidosomes thermogram with kaolin shell reinforced with fatty acids and spray-dried (Example 4). 
     
    
    
     DETAILED DESCRIPTION OF THE INVENTION 
     This invention describes a process of making colloidosomes-type microcapsules of liquid phases insoluble in water, using nanoparticles, microparticles or their combination, superficially modified obtained by ionic gelation. 
     To obtain the colloidosome, an oil-in-water (O/W) emulsion is initially generated stabilized with nano- or solid microparticles insoluble in water and obtained by ionic gelation. The emulsion stabilized with the particles is produced by applying to the disruptive forces system such as shear, cavitation, collision between particles, pressure drop or the combination of two or more types of these disruptive forces. 
     Subsequently, the particles are fixed to the interface by polyelectrolytes adsorption, heat treatment, cross-linking, treatment with emulsion of a saturated fatty acid or fatty acids mixture, generating the colloidosome with the insoluble phase in water encapsulated in the core and coated by the shell particles. The colloidosomes obtained can have sizes between 100 nm and several millimeters. The mechanical stability of colloidosomes obtained in suspension makes it possible to carry out drying processes to achieve colloidosomes in the form of dry powder. 
     The process of ionic gelation on particles is achieved by polyvalent cations addition to an insoluble solids suspension, previous sub-environment thermal treatment that allows a controlled gelation of the charged macromolecules. The subsequent increase in temperature consolidates the shell formation over the insoluble solid particles. The detailed characteristics and conditions of the ionic gelation process are described in the document CO2013203104, which is incorporated in its entirety as a reference. 
     The suspension of solid particles treated superficially by ionic gelation is subsequently dried, generating particles agglomerates. These agglomerates are redispersed in water and subjected to a milling process, to generate individual particles that are used to emulsify a liquid phase insoluble in water by applying the mentioned disruptive forces. 
     The emulsion stabilized with the particles, requires the particles fixation in the liquid interface of the drops by polyelectrolytes adsorption, heat treatment, cross-linking, treatment with emulsion of a saturated fatty acid or fatty acids mixture, generating a colloidosome, where the insoluble phase in water corresponds to the core and the particles form the shell. 
     The colloidosomes typical diameters are between 100 nm and several millimeters. The wet system produces typically spherical colloidosomes with a liquid phase content insoluble in water between a 5% and 80%. The colloidosomes in aqueous suspension can be subsequently dried by spray drying, lyophilization, tray heating or other alternative drying methods. The polyelectrolytes adsorption, crosslinking or particles sintering degree, determines the shell mechanical properties, and consequently, its stability in the drying processes. Typically, colloidosomes can be dried at temperatures up to 250° C. without causing rupture of their shell. 
     The liquid phases colloidosomes elaboration process insoluble in water using particles superficially modified by ionic gelation of this invention, comprises the following steps:
         a) develop a negatively charged polyelectrolytes solution, adjust the pH at a certain value between 5.0 and 8.0 and cool;   b) elaborate a suspension of insoluble solids in water, adjusting its pH;   c) mix the polyelectrolytes solution obtained in a) with the suspension of water-insoluble solids obtained in b), applying agitation to the system and controlling its temperature;   d) add a solution of polyvalent ions to the aqueous suspension obtained in c) and heat;   e) dry the aqueous suspension until get an agglomerated solid particles dry powder superficially modified;   f) redisperse the dry solid particles in water by applying disruptive forces to prevent the agglomerates formation;   g) emulsify an insoluble liquid in water using as emulsifier the suspension obtained in (f) and optionally bring the system temperature to more than 40° C.;   h) add a polyvalent cations solution to the emulsion stabilized by particles obtained in step (g);   i) add a solution of a polyelectrolyte negatively charged to the emulsion stabilized by particles obtained in step (h) generating colloidosomes in suspension;   j) repeat steps (h) and (i) to generate shells with greater mechanical stability;   k) optionally adding the emulsion stabilized by particles obtained in step (g) to an emulsion of saturated fatty acids to generate shells of greater mechanical stability; and   l) optionally drying the suspended colloidosome from step (j) or (k) to get powder colloidosomes. Optional drying of the colloidosome obtained in step (j) or (k) can be done in the presence of polyelectrolytes.       

     The characteristics of the colloidosomes elaboration process as well as the characteristics of the colloidosomes generated by said process are described in detail. These characteristics can be interchanged to describe both the process and the colloidosome. 
     Water insoluble solids are nanoparticles, microparticles or their combination, which preferably must generate surface charge when dispersed in water 
     as a product of dissociation of their functional groups when interacting with water or another protic solvent. In a preferred modality, the insoluble solids for the ionic gelation process are metallic and non-metallic minerals or other insoluble solids such as phyllosilicates, polymer particles and insoluble solids obtained via synthesis, extraction or by bioprocesses. 
     The particle diameter of the water insoluble solid suitable for the ionic gelation process is between 100 nm and several millimeters. Compact solid particles are preferred for the surface modification by ionic gelation process, although other morphologies can also be employed. The solids concentration in the system is usually lower than 80%. 
     The shell-forming polyelectrolytes are typically proteins, polysaccharides or synthetic polymers negatively charged. In a preferred modality, the proteins include milk proteins, vegetable origin proteins, gelatin, albumins and mixtures thereof. Salts of these proteins such as sodium caseinate and calcium caseinate can also be used. Useful polysaccharides to be used for solids surface modification include hydrocolloids such as gum arabic, xanthan, alginate salts, cellulose derivatives, pectin salts, carrageenans, guar gum and mixtures thereof. 
     It is necessary to achieve an adequate hydration and interaction between the macromolecules and the surface of the solid insoluble in water, for which it is convenient to lower the system temperature at temperatures below 10° C. To induce the polyelectrolytes ionic gelation on the surface of the insoluble solid, a source of polyvalent cations is added to the solids suspension in the presence of the macromolecules. The source of polyvalent cations is preferably a soluble salt or slightly insoluble in water. In a preferred modality, the polyvalent cations source is a CaCl 2 ) solution at a concentration no greater than 2 molar. 
     Once the macromolecules adsorption process on the surface of the insoluble solid is carried out, the temperature of the system is increased to induce its ionic gelation, which is achieved at temperatures close to 25° C., although in some cases an increase in the temperature of the system of up to 80° C. may be required. The colloidosome suspension is subsequently dried, preferably by spray drying, to generate agglomerated and superficially modified dry solid particles by the ionic gelation process. 
     The agglomerates of solid particles are redispersed in water, to then generate individual particles by applying disruptive forces such as shear or pressure changes, without excluding others such as cavitation or particles collision. The aqueous suspension of deagglomerated solid particles is used as the emulsifying system of a water insoluble liquid with the application of these same disruptive forces. 
     The generated emulsion is stabilized by the adsorbed particles in the liquid interface, which confer colloidal and mechanical stability to the emulsion drops. The typical size of emulsion drops is ten times or more the diameter of the stabilizing particle and the emulsified oil ratio: stabilizing particles are between 0.1:1 and 5:1. 
     Once the emulsion stabilized by particles is generated, the solid particles are fixed to generate a stable shell and, consequently, to produce the colloidosome. For this, a solution of a negatively charged polyelectrolyte previously hydrated is added and, if necessary, cooled to temperatures below 10° C. Subsequently, the formation of a stabilizing film of the particles adsorbed on the emulsion drops is induced by the addition of a divalent cations source. 
     It is also possible to make the particles fixation by the addition of cross-linking agents, heat treatment, coacervation or other methods. In a preferred modality, the cross-linking agent is selected from the group consisting of alcohols, aldehydes (e.g., glutaraldehyde), vinyls, ketones, proteins and enzymes (e.g., transglutaminase). The particles fixation by heat treatment should induce flocculation or coalescence of the particles on the interface of the emulsion drops. 
     Once the particles are fixed on the liquid interface, the colloidosomes are dried to generate a colloidosomes dry powder, preferably by spray drying. Other methods such as lyophilization or tray drying are equally viable for the colloidosomes drying. Depending on the conditions of the drying process, individual colloidosomes or colloidosomes agglomerates can be generated. The colloidosomes mechanical stability in the aqueous dispersion makes it possible to preserve the shell integrity during the drying, generating colloidosomes in the form of powder with recovery efficiency greater than 70%. 
     The following examples illustrate the invention, without the inventive concept being restricted thereto. 
     EXAMPLES 
     Example 1. Oleic Acid Colloidosomes Elaboration with Micrometric-Sized Calcium Carbonate Particles 
     540 g of a sodium caseinate solution (5% w/w) were prepared by hydration for more than 2 hours and cooling to 5° C. and adjusting the pH to 6.5. In parallel, 412 g of a calcium carbonate suspension (67.0% w/w were prepared and average particle size of 3 μm), pH was adjusted to 6.5 and cooled to 5° C. This suspension was mixed with the sodium caseinate solution by mechanical agitation. 
     Subsequently, 48 g of a calcium chloride solution (4.1% w/w) were added, the temperature of the whole system was increased to 25° C. and dried in a spray dryer at an inlet temperature of 200° C., suction of 40 m 3 /h, feed pump to 8 mL/min and an incoming air flow to 1200 L/h, to obtain microencapsulated calcium carbonate. 
     The calcium carbonate dry agglomerated particles were redispersed in water to form a suspension to 30% (w/w) which was subsequently milled in a pearl mill for at least 30 minutes, thus achieving the calcium carbonate agglomerates destruction.  FIG. 1  shows the change in particle size of the agglomerated system and the ground system in suspension. 
     To elaborate the emulsion stabilized with particles, 100 g of the calcium carbonate suspension (30%) were taken of deagglomerated particles, mixed with 15 g of oleic acid, and diluted with water to obtain a system with a 70% of water. This system was homogenized by applying high shear using a rotor stator equipment to 15000 rpm for 10 minutes, after which they were added 0.3 g of CaCl 2 ). 
     The particles adsorption on the liquid interface was corroborated by optical microscopy ( FIG. 2 ), where the formation of emulsion drops stabilized with calcium carbonate microparticles is evidenced. The particle size distribution of this system is shown in  FIG. 3 . Then, 100 g of the emulsion stabilized with particles were taken and 3 g of sodium caseinate were added, stirred for 2 hours and cooled to 5° C. Finally, 0.22 g of NaCl were added and the system temperature was increased up to 25° C. The colloidosomes suspension was spray-dried at an inlet temperature of 180° C., suction of 30 m 3 /h, feed pump with speed of 6 mL/min and incoming air flow of 1000 L/h. 
     The dry colloidosomes formation was evidenced in the scanning electron microphotographs of  FIG. 4 . The particle size distribution is presented in  FIG. 5 . The oleic acid content was corroborated by thermogravimetric analysis ( FIG. 6 ), where an encapsulated content in the colloidosome of approximately 28% was observed. 
     Example 2. Sunflower Oil Colloidosomes Elaboration with Titanium Dioxide Particles of Sub-Micrometric Size 
     200 g of titanium dioxide were dispersed in 200 g of a sodium caseinate solution (0.5% w/w), using a cowles-type disperser for 32 minutes at 5° C. To this suspension was added 341 g of a sodium caseinate solution (6.22% w/w) hydrated previously for more than 2 hours and maintaining the temperature of 5° C. and pH adjusted in 6.5, to generate a system with a solids concentration of 30%. 
     From the previous suspension, 200 g were taken and 32.8 g of a CaCl 2 ) (5.0% w/w) solution, the system temperature was increased 5° C. to 25° C. and dried in a spray dryer with an inlet temperature of 200° C., suction of 40 m 3 /h, feeding pump with a flow of 8 mL/min and incoming air flow of 1200 L/h, to so obtain microencapsulated titanium dioxide. 
     The dried agglomerated particles of titanium dioxide were redispersed in water to form a suspension at 30% (w/w), which was later milled in a pearl mill for at least 30 minutes to destroy the titanium dioxide agglomerates.  FIG. 7  shows the agglomerated system particle size change and of the milled system. 
     To make the emulsion stabilized with particles, 100 g of the titanium dioxide suspension (30%) were taken of deagglomerated particles, mixed with 30 g of sunflower oil and the system was homogenized by applying high pressure (1000 bar). The particles adsorption on the liquid interface was corroborated by optical microscopy ( FIG. 8 ), where the emulsion drops formation stabilized with titanium dioxide microparticles is evidenced. The particle size distribution of this system is shown in  FIG. 9 . 
     Subsequently, 100 g of the sunflower oil emulsion stabilized with particles were taken and 3 g of sodium caseinate were added. The colloidosomes suspension was spray-dried at an inlet temperature of 180° C., a suction of 30 m 3 /h, feed pump with speed of 6 mL/min and an incoming air flow of 1000 L/h. 
     The dry colloidosomes formation could be evidenced by the scanning electron microphotographs shown in  FIG. 10 , while the particle size distribution is shown in  FIG. 11 . The content of sunflower oil was corroborated by thermogravimetric analysis ( FIG. 12 ), where sunflower oil encapsulated content in the colloidosome close to 32% was observed. 
     Example 3. Colloidosomes Elaboration of a Mono and Diglycerides Mixture with Micrometric Calcium Phosphate Particles 
     540 g of a sodium caseinate solution (5% w/w) were prepared by hydration for more than 2 hours and pH adjustment to 6.5. In parallel, 412 g of a calcium phosphate suspension (67% w/w and average particle size of 3 μm) were prepared and with adjustment of pH to 6.5, which was mixed with the sodium caseinate solution while maintaining mechanical agitation. 
     Subsequently, 48 g of a CaCl 2 ) (4.1% w/w) solution to 25° C. were dried and the mixture was dried in a spray dryer at an inlet temperature of 200° C., suction of 40 m 3 /h, feed pump with speed 8 mL/min and an incoming air flow of 1200 L/h, to obtain microencapsulated calcium phosphate. 
     The calcium phosphate dry agglomerated particles were redispersed in water to obtain a suspension at 25% (w/w), which was then milled in a pearl mill for at least 30 minutes to destroy the calcium phosphate agglomerates.  FIG. 13  shows the change in particle size of the agglomerated system and the milled system. 
     To make the emulsion stabilized with particles, 100 g of the calcium carbonate suspension (25%) of deagglomerated particles were taken and heated up to 55° C. In parallel way 25 g of a mono and diglycerides mixture were heated up to 55° C. and were added slowly to the suspension of deagglomerated particles by applying stirring to 15000 rpm using a rotor-stator equipment. 
     The particles adsorption on the liquid interface was corroborated by optical microscopy ( FIG. 14 ), where the emulsion drops formation stabilized with calcium phosphate microparticles is evidenced. The particle size distribution of this system is presented in  FIG. 15 . Finally, 50 g of a 10% sodium caseinate solution (w/w) and 8.0 g of a CaCl 2  dihydrate to 5% (w/w) solution. 
     The colloidosomes suspension is spray-dried at an inlet temperature of 180° C., suction of 30 m 3 /h, feeding pump with speed of 6 mL/min and incoming air flow of 1000 L/h. The dry colloidosomes formation was evidenced in the scanning electron microphotographs ( FIG. 16 ), with a particle size distribution presented in  FIG. 17 . The mono and diglyceride content was corroborated by thermogravimetric analysis ( FIG. 18 ), where an encapsulated material content close to 37% was observed. 
     Example 4. Canola Oil Colloidosomes Elaboration with Particles of Micrometric Kaolin and Reinforced with Fatty Acids 
     500 g of a suspension were prepared by mixing 200 g of kaolin with 295 g of sodium caseinate solution in a cowles-type disperser at 1000 rpm. To the resulting suspension having 40% of solid material, the pH was adjusted to 6.5 and 166.7 g of a sodium caseinate solution (9% w/w) were added with stirring and 5° C. during 2 hours and 30 g of CaCl 2 ) dihydrate (5% w/w) solution maintaining 5° C. of temperature. 
     Subsequently, the temperature of the entire system was increased to 25° C. and dried in a spray dryer at an incoming temperature of 200° C., suction of 40 m 3 /h, feed pump to 8 mL/min and incoming air flow at 1200 L/h. 30 g of solid material obtained by spray drying were taken, dispersed in 70 g of water and stirred for 2 minutes at 15000 rpm using a rotor-stator equipment. 
       FIG. 19  shows the change in particle size of the agglomerated system and the milled system. 
     The obtained system was diluted by adding 85 g of water and then 60 g of canola oil were dispersed using a rotor-stator type disperser at 12000 rpm maintaining the stirring for two minutes after the total oil addition. The particles adsorption on the liquid interface was corroborated by optical microscopy ( FIG. 20 ), where the formation of emulsion drops stabilized with kaolin microparticles is evidenced. The particle size distribution of this system is presented in  FIG. 21 . 
     To the colloidosomes suspension were added 55 g of sodium caseinate solution (10% w/w) applying stirring at 500 rpm and heating the system between 40° C. and 50° C. for 5 minutes. Then, this system was cooled to 5° C. maintaining the agitation to then adding 8 g of CaCl 2 ) solution dihydrated. The system temperature was increased to 25° C. and spray-dried at an inlet temperature of 180° C., suction 30 m 3 /h, feeding pump with a speed of 6 mL/min and an incoming air flow at 1000 L/h. The dried product is made up of canola oil drops coated with kaolin particles, as shown in the SEM photomicrographs ( FIG. 22 ). 
     Alternatively, canola oil colloidosomes with a kaolin particle shell can be reinforced with a fatty shell. For this, an emulsion was prepared using 20 g of a 1:1 mixture by mass of stearic acid/palmitic acid and 180 g of water, homogenized at a temperature above 70° C. for 5 minutes with a rotor-stator disperser (12000 rpm). 
     100 g of a colloidosomes suspension were taken with 110 g of fatty acids emulsion at 70° C. and by mechanical stir they were dispersed with a disperser (500 rpm) and then cooled to room temperature. Colloidosomes coated with fatty compounds are evidenced in the optical micrograph of  FIG. 23 . 
     This system is subsequently brought to a temperature of 5° C., to then continue with the process described above to obtain dry microcapsules reinforced with a layer of saturated fat, as can be seen in the SEM photomicrograph of  FIG. 24 . The content of canola oil was determined by thermogravimetric analysis ( FIG. 25 ), where a content of encapsulated material close to 36% was observed. 
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