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Dragon's Blood Sap Microencapsulation within Whey Protein Concentrate and Zein Using Electrospraying Assisted by Pressurized Gas Technology

RiuNet. Repositorio Institucional de la Universitat Politécnica de Valéncia
  • Escobar-García, Juan David
  • Prieto, Cristina
  • Pardo-Figuerez, Maria
  • Lagaron, Jose M.
[EN] Dragon's blood sap (DBS) obtained from the bark of Croton lechleri (Müll, Arg.) is a complex herbal remedy of pharmacological interest due to its high content in polyphenols, specifically proanthocyanidins. In this paper, electrospraying assisted by pressurized gas (EAPG) was first compared with freeze-drying to dry natural DBS. Secondly, EAPG was used for the first time to entrap natural DBS at room temperature into two different encapsulation matrices, i.e., whey protein concentrate (WPC) and zein (ZN), using different ratios of encapsulant material: bioactive compound, for instance 2:1 w/w and 1:1 w/w. The obtained particles were characterized in terms of morphology, total soluble polyphenolic content (TSP), antioxidant activity, and photo-oxidation stability during the 40 days of the experiment. Regarding the drying process, EAPG produced spherical particles with sizes of 11.38 ± 4.34 µm, whereas freeze-drying produced irregular particles with a broad particle size distribution. However, no significant differences were detected between DBS dried by EAPG or freeze-drying in TSP, antioxidant activity, and photo-oxidation stability, confirming that EAPG is a mild drying process suitable to dry sensitive bioactive compounds. Regarding the encapsulation process, the DBS encapsulated within the WPC produced smooth spherical microparticles, with average sizes of 11.28 ± 4.28 µm and 12.77 ± 4.54 µm for ratios 1:1 w/w and 2:1 w/w, respectively. The DBS was also encapsulated into ZN producing rough spherical microparticles, with average sizes of 6.37 ± 1.67 µm and 7.58 ± 2.54 µm for ratios 1:1 w/w and 2:1 w/w, respectively. The TSP was not affected during the encapsulation process. However, a slight reduction in antioxidant activity measured by DPPH was observed during encapsulation. An accelerated photo-oxidation test under ultraviolet light confirmed that the encapsulated DBS showed an increased oxidative stability in comparison with the non-encapsulated DBS, with the stability being enhanced for the ratio of 2:1 w/w. Among the encapsulating materials and according to the ATR-FTIR results, ZN showed increased protection against UV light. The obtained results demonstrate the potential of EAPG technology in the drying or encapsulation of sensitive natural bioactive compounds in a continuous process available at an industrial scale, which could be an alternative to freeze-drying., This research was funded by the Valencian Innovation Agency (AVI) BIOENCAP project (reference number INNCAD00-18-31), H2020 EU FODIAC project (reference number 778388), and the H2020 EU projects CAPSULTEK (reference number 873827), grant PID2021-128749OB-C31 funded by MCIN/AEI/10.13039/501100011033 and by ERDF A way of making Europe. J.D.E.G. would like to thank the Spanish Ministry of Economy, Industry and Competitivity for his predoctoral grant (DI-6906). C.P. would like to thank the Valencian Ministry of Innovation, Universities, Science and Digital Society for her postdoctoral grant (CIAPOS/2021/45).




Room Temperature Nanoencapsulation of Bioactive Eicosapentaenoic Acid Rich Oil within Whey Protein Microparticles

RiuNet. Repositorio Institucional de la Universitat Politécnica de Valéncia
  • Escobar-García, Juan David
  • Prieto, Cristina
  • Pardo-Figuerez, Maria
  • Lagaron, José M.
[EN] In this study, emulsion electrospraying assisted by pressurized gas (EAPG) has been performed for the first time to entrap ca. 760 nm droplets of the bioactive eicosapentaenoic acid (EPA)-rich oil into whey protein concentrate (WPC) at room temperature. The submicron droplets of EPA oil were encapsulated within WPC spherical microparticles, with sizes around 5 µm. The EPA oil did not oxidize in the course of the encapsulation performed at 25 °C and in the presence of air, as corroborated by the peroxide value measurements. Attenuated Total Reflection-Fourier Transform Infrared spectroscopy and oxygen consumption tests confirmed that the encapsulated EPA-rich oil showed increased oxidative stability in comparison with the free oil during an accelerated oxidation test under ultraviolet light. Moreover, the encapsulated EPA-rich oil showed increased thermal stability in comparison with the free oil, as measured by oxidative thermogravimetric analysis. The encapsulated EPA-rich oil showed a somewhat reduced organoleptic impact in contrast with the neat EPA oil using rehydrated powdered milk as a reference. Finally, the oxidative stability by thermogravimetric analysis and organoleptic impact of mixtures of EPA and docosahexaenoic acid (DHA)-loaded microparticles was also studied, suggesting an overall reduced organoleptic impact compared to pure EPA. The results here suggest that it is possible to encapsulate 80% polyunsaturated fatty acids (PUFAs)-enriched oils by emulsion EAPG technology at room temperature, which could be used to produce personalized nutraceuticals or pharmaceuticals alone or in combination with other microparticles encapsulating different PUFAs to obtain different targeted health and organoleptic benefits., This research was funded by the MICIU (RTI-2018-097249B-C1), EU H2020-MSCA-RISE2017-778388-FODIAC and the EU H2020-EIC-SMEinst-2018-2020-SME instrument-873827-CAPSULTEK, CDTI-CIEN DANTIAN-IDI-20190954 and the CYTED thematic network (319RT0576). J.D.E.-G. wants to thank the Spanish Ministry of Economy, Industry and Competitivity for his predoctoral grant (DI-6906).




Nanodroplets of Docosahexaenoic Acid-Enriched Algae Oil Encapsulated within Microparticles of Hydrocolloids by Emulsion Electrospraying Assisted by Pressurized Gas

Digital.CSIC. Repositorio Institucional del CSIC
  • Prieto López, Cristina
  • Lagarón Cabello, José María
© 2020 by the authors., Long chain polyunsaturated omega-3 fatty acids (PUFAs), namely eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), are important functional ingredients due to their well-documented health benefits, but highly susceptible to oxidation. One of the most promising approaches to preserve bioactives is their encapsulation within protective matrices. In this paper, an innovative high throughput encapsulation technique termed as emulsion electrospraying assisted by pressurized gas (EAPG) was used to encapsulate at room temperature nanodroplets of algae oil into two food hydrocolloids, whey protein concentrate and maltodextrin. Spherical encapsulating particles with sizes around 5 µm were obtained, where the oil was homogeneously distributed in nanometric cavities with sizes below 300 nm. Peroxide values under 5 meq/kg, demonstrated that the oil did not suffer from oxidation during the encapsulation process carried out at room temperature. An accelerated stability assay against oxidation under strong UV light was performed to check the protective capacity of the different encapsulating materials. While particles made from whey protein concentrate showed good oxidative stability, particles made from maltodextrin were more susceptible to secondary oxidation, as determined by a methodology put forward in this study based on ATR-FTIR spectroscopy. Further organoleptic testing performed with the encapsulates in a model food product, i.e., milk powder, suggested that the lowest organoleptic impact was seen for the encapsulates made from whey protein concentrate. The obtained results demonstrate the potential of the EAPG technology using whey protein concentrate as the encapsulating matrix, for the stabilization of sensitive bioactive compounds., This research was funded by the Spanish Ministry of Science and Universities (project RTI-2018-097249-B-C21), the Valencian Innovation Agency (AVI) BIOENCAP project (reference number INNCAD00-18-31), H2020 EU FODIAC project (reference number 778388) and the H2020 EU projects CAPSULTEK (reference number 873827) and YPACK (reference number 773872)., Peer reviewed




Encapsulation of β-Carotene by Emulsion Electrospraying Using Deep Eutectic Solvents

Digital.CSIC. Repositorio Institucional del CSIC
  • Basar, Ahmet Ozan
  • Prieto López, Cristina
  • Durand, Erwann
  • Villeneuve, Pierre
  • Sasmazel, Hilal Turkoglu
  • Lagarón Cabello, José María
© 2020 by the authors., The encapsulation β-carotene in whey protein concentrate (WPC) capsules through the emulsion electrospraying technique was studied, using deep eutectic solvents (DES) as solvents. These novel solvents are characterized by negligible volatility, a liquid state far below 0 °C, a broad range of polarity, high solubilization power strength for a wide range of compounds, especially poorly water-soluble compounds, high extraction ability, and high stabilization ability for some natural products. Four DES formulations were used, based on mixtures of choline chloride with water, propanediol, glucose, glycerol, or butanediol. β-Carotene was successfully encapsulated in a solubilized form within WPC capsules; as a DES formulation with choline chloride and butanediol, the formulation produced capsules with the highest carotenoid loading capacity. SEM micrographs demonstrated that round and smooth capsules with sizes around 2 µm were obtained. ATR-FTIR results showed the presence of DES in the WPC capsules, which indirectly anticipated the presence of β-carotene in the WPC capsules. Stability against photo-oxidation studies confirmed the expected presence of the bioactive and revealed that solubilized β-carotene loaded WPC capsules presented excellent photo-oxidation stability compared with free β-carotene. The capsules developed here clearly show the significant potential of the combination of DES and electrospraying for the encapsulation and stabilization of highly insoluble bioactive compounds., This research was funded by the COST Action FP1405 (STSM reference number 41084), the MICIU project (RTI-2018-097249-B-C21), the Valencian Innovation Agency (AVI) BIOENCAP project (reference number INNCAD00-18-31), the H2020 EU FODIAC project (reference number 778388), and the H2020 EU projects CAPSULTEK (reference number 873827)., Peer reviewed




Nanostructured Valsartan Microparticles with Enhanced Bioavailability Produced by High-Throughput Electrohydrodynamic Room-Temperature Atomization

Digital.CSIC. Repositorio Institucional del CSIC
  • Prieto López, Cristina
  • Evtoski, Zoran
  • Pardo Figuérez, María
  • Hrakovsky, Julia
  • Lagarón Cabello, José María
The high-throughput drying and encapsulation technique called electrospraying assisted by pressurized gas (EAPG) was used for the first time to produce nanostructured valsartan within microparticles of excipients. Valsartan, a poorly absorbed and lipid-soluble drug, was selected since it is considered a good model for BCS class II drugs. Two different polymeric matrices were selected as excipients, i.e., hydroxypropyl methylcellulose (HPMC) and lactose monohydrate, while Span 20 was used as a surfactant. The produced 80% valsartan loading formulations were characterized in terms of morphology, crystallinity, in vitro release, in vitro Caco-2 cells’ permeability, and in vivo pharmacokinetic study. Spherical microparticles of ca. 4 μm were obtained within which valsartan nanoparticles were seen to range from 150 to 650 nm. Wide-angle X-ray scattering and differential scanning calorimetry confirmed that valsartan had a lower and/or more ill-defined crystallinity than the commercial source, and photon correlation spectroscopy and transmission electron microscopy proved that it was dispersed and distributed in the form of nanoparticles of controlled size. In vitro dissolution tests showed that the HPMC formulation with the lowest API particle size, i.e., 150 nm, dissolved 2.5-fold faster than the commercial valsartan in the first 10 min. This formulation also showed a 4-fold faster in vitro permeability than the commercial valsartan and a 3-fold higher systemic exposure than the commercial sample. The results proved the potential of the EAPG processing technique for the production of safe-to-handle microparticles containing high quantities of a highly dispersed and distributed nanonized BCS class II model drug with enhanced bioavailability., This research was funded by the Spanish Ministry of Science and Universities (project RTI-2018-097249-B-C1), the Valencian Innovation Agency BIOENCAP project (reference no. INNCAD00-18-31), the H2020 EU projects FODIAC (reference no. 778388), the CAPSULTEK (reference no. 873827), and the CYTED thematic network (reference no. 319RT0576)., Peer reviewed




Room Temperature Nanoencapsulation of Bioactive Eicosapentaenoic Acid Rich Oil within Whey Protein Microparticles

Digital.CSIC. Repositorio Institucional del CSIC
  • Escobar García, Juan D.
  • Prieto López, Cristina
  • Pardo Figuérez, María
  • Lagarón Cabello, José María
In this study, emulsion electrospraying assisted by pressurized gas (EAPG) has been performed for the first time to entrap ca. 760 nm droplets of the bioactive eicosapentaenoic acid (EPA)-rich oil into whey protein concentrate (WPC) at room temperature. The submicron droplets of EPA oil were encapsulated within WPC spherical microparticles, with sizes around 5 µm. The EPA oil did not oxidize in the course of the encapsulation performed at 25 °C and in the presence of air, as corroborated by the peroxide value measurements. Attenuated Total Reflection—Fourier Transform Infrared spectroscopy and oxygen consumption tests confirmed that the encapsulated EPA-rich oil showed increased oxidative stability in comparison with the free oil during an accelerated oxidation test under ultraviolet light. Moreover, the encapsulated EPA-rich oil showed increased thermal stability in comparison with the free oil, as measured by oxidative thermogravimetric analysis. The encapsulated EPA-rich oil showed a somewhat reduced organoleptic impact in contrast with the neat EPA oil using rehydrated powdered milk as a reference. Finally, the oxidative stability by thermogravimetric analysis and organoleptic impact of mixtures of EPA and docosahexaenoic acid (DHA)-loaded microparticles was also studied, suggesting an overall reduced organoleptic impact compared to pure EPA. The results here suggest that it is possible to encapsulate 80% polyunsaturated fatty acids (PUFAs)-enriched oils by emulsion EAPG technology at room temperature, which could be used to produce personalized nutraceuticals or pharmaceuticals alone or in combination with other microparticles encapsulating different PUFAs to obtain different targeted health and organoleptic benefits., This research was funded by the MICIU (RTI-2018-097249B-C1), EU H2020-MSCA-RISE-2017-778388-FODIAC and the EU H2020-EIC-SMEinst-2018-2020-SME instrument-873827-CAPSULTEK, CDTI-CIEN DANTIAN-IDI-20190954 and the CYTED thematic network (319RT0576)., Peer reviewed




Bioavailability enhancement of nanostructured microparticles of carvedilol

Digital.CSIC. Repositorio Institucional del CSIC
  • Prieto López, Cristina
  • Evtoski, Zoran
  • Pardo Figuérez, María
  • Lagarón Cabello, José María
A high throughput encapsulation technique termed as electrospraying assisted by pressurized gas technology (EAPG) was used to produce nano-within-micro structures of a Biopharmaceutics Classification System (BCS) Class II model drug. Carvedilol is a lipid soluble compound, poorly absorbed in the gastrointestinal tract. The produced formulations were characterized in terms of morphology, crystallinity, in vitro dissolution test, in vitro Caco-2 cells permeability and in vivo pharmacokinetics in rats. Spherical microparticles with a carvedilol loading of 80% were produced with sizes around 4 μm. DLS and TEM suggested that carvedilol is released in the form of nanoparticles of controlled size when the microparticles are put in solution, and WAXS and DSC confirmed that carvedilol was in an amorphous state. In vitro dissolution tests showed that the produced microparticles dissolved 4-fold faster than the commercial carvedilol in the first 30 min. The apparent permeability in Caco-2 cells of the produced formulations was approximately 2.5-fold higher than the apparent permeability of the commercial carvedilol. The preliminary pharmacokinetic assay suggested a reduction in 2 h of the Cmax for the prepared formulations, but due to the high variability observed, the results need to be confirmed in further studies. This work showed the potential of nanostructured microparticles of an API via EAPG to increase dissolution rate and hence the bioavailability of a BCS Class II drug., This research was funded by the Spanish Ministry of Science and Universities (project RTI-2018-097249-B-C21), the Valencian Innovation Agency BIOENCAP project (reference number INNCAD00-18-31), the H2020 EU projects FODIAC (reference number 778388) CAPSULTEK (reference number 873827) and the CYTED thematic network (code 319RT0576)., Peer reviewed




CAPSULTEK® Nº. 873827 - Unlocking the potential of thermo-sensitive bioactive ingredients through a disruptive encapsulation method.

Digital.CSIC. Repositorio Institucional del CSIC
  • Talón Argente, Emma
  • Palau Aloy, José Luis
  • Markwort, Lars
The dataset is made available under the Open Database License. Any rights in individual contents of the database are licensed under the Database Contents License. Please, read the full ODbL 1.0 license text for the exact terms that apply. Users of the dataset are free to: Share: copy, distribute and use the database, either commercially or non-commercially. Create: produce derivative works from the database. Adapt: modify, transform and build upon the database. Under the following conditions: Attribution: You must attribute any public use of the database, or works produced from the database. For any use or redistribution of the database, or works produced from it, you must make clear to others the license of the original database. Share-Alike: If you publicly use any adapted version of this database, or works produced from an adapted database, you must also offer that adapted database under the ODbL., The objective of the Capsultek project is to scale-up the electrospraying assisted by pressurized gas (EAPG) technology for microencapsulation and drying of thermolabile active ingredients to subsequently validate it, reaching a production capacity of 10 tons/year in our facilities in Spain.
Bearing in mind the aim of the project, the information related to the Data Management Plan (DMP), data collection registry, is provided.
In relation to the data collection, the purpose was to analyze the real performance of our equipment and to improve our process for the up-scale phase. The data generated are related with temperatures, flowrates and other variables to demonstrate that we can accomplish our objectives., CAPSULTEK has received funding from the European Union ́s Horizon 2020 research and innovation programme under grant agreement Nº 873827, Peer reviewed
Proyecto: EC/H2020/873827




Development of Multilayer Ciprofloxacin Hydrochloride Electrospun Patches for Buccal Drug Delivery

Digital.CSIC. Repositorio Institucional del CSIC
  • Teno, Jorge
  • Pardo Figuérez, María
  • Figueroa López, Kelly J.
  • Prieto López, Cristina
  • Lagarón Cabello, José María
Bacterial infections in the oral cavity can become a serious problem causing pain, sores and swelling for several weeks. This type of infection could be alleviated using mucoadhesive delivery systems, allowing local administration of the antibiotic to inhibit bacterial spreading. This work reports the development of a multilayer antibiotic patch containing ciprofloxacin hydrochloride (CPX)-loaded electrospun fibers for the treatment of such infections. For this, the release kinetics of the CPX-loaded fibers was modulated using different ratios of polyester blends. The selected reservoir layer was analyzed by scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), wide angle x-ray scattering (WAXS) and differential scanning calorimetry (DSC). These analyses confirmed the presence and good distribution of the drug in the fibers and that the drug is in an amorphous state within the reservoir layer. To enhance mucoadhesion whilst ensuring drug directionality, the reservoir layer was assembled to a backing and an adhesive layer. This multilayer patch was assessed in terms of in vitro drug release, adhesion and antimicrobial properties. The multilayer strategy showed excellent antimicrobial properties over time and also a strong adhesion patch time in the volunteers for an average of 7 h. These results highlight the capabilities of multilayer electrospun patches as platforms to treat oral infections., This research was funded by the Spanish Ministry of Science and Universities (project RTI-2018-097249-B-C21), the Valencian Innovation Agency BIOENCAP project (reference number INNCAD00-18-31), the H2020 EU projects FODIAC (reference number 778388), CAPSULTEK (reference number 873827), the CYTED thematic network (code 319RT0576) and BIOINICIA’s internal project ASOPHARM., Peer reviewed




Comparison of the Stability of a Camu Camu Extract Dried and Encapsulated by Means of High-Throughput Electrospraying Assisted by Pressurized Gas

Digital.CSIC. Repositorio Institucional del CSIC
  • Escobar-García, Juan David
  • Prieto López, Cristina
  • Talón Argente, Emma
  • Lagarón Cabello, José María
This study explores the impact on the stability of drying and the encapsulation of a camu camu extract (CCX) using the non-thermal, high-throughput electrospraying assisted by pressurized gas (EAPG) technique. The dried and encapsulated products by the EAPG processing techniques were compared in terms of total soluble phenolic compounds, antioxidant activity, and storage stability. Whey protein concentrate (WPC) and zein (ZN) were selected as the protective excipients for encapsulation. Dried and encapsulated products were obtained in the form of microparticles, which were smaller and more spherical in the case of the encapsulates. No significant differences were observed in the total polyphenolic content (TSP), and only relatively small differences in the antioxidant capacity were measured among samples. The generated products were subjected to various storage conditions to assess their stability and the preservation of the TSP and the antioxidant properties, i.e., 0% relative humidity (RH) and 4 °C; 0% RH and 21 °C; 23% RH and 21 °C; 56% RH and 21 °C; and UV light exposure. The results indicated that ZN encapsulation notably enhanced the retention of total soluble polyphenols and the antioxidant activity compared to WPC and dried CCX, especially in the ratio of 2:1 (encapsulating polymer: dried CCX). This study demonstrates the potential of protein-based encapsulation, particularly using ZN, for stabilizing bioactive compounds against degradation mechanisms induced by humidity, temperature, or ultraviolet radiation exposure., This research was funded by the Valencian Innovation Agency (AVI) BIOENCAP project (reference number INNCAD00-18-31), the H2020 EU FODIAC project (reference number 778388), and the H2020 EU projects CAPSULTEK (reference number 873827), grant PID2021-128749OB-C31 funded by MCIN/AEI/10.13039/501100011033, and by ERDF A Way of Making Europe., Peer reviewed