BioaccessibilityEvaluation by In Vitro Digestion of MicroencapsulatedExtracts of Habanero Pepper Leaves Obtained Through an Optimized Spray-DryingProcess

Abstract

Habanero pepper (Capsicum chinense Jacq.)leaves, a major by-product of pepper cultivation in the Yucatán Peninsula, arean underexploited source of phenolic compounds with relevant antioxidantpotential. In this work, phenolic-rich extracts obtained with a cholinechloride-glucose Natural Deep Eutectic Solvent (NADES) and ultrasound-assistedextraction were microencapsulated by spray-drying using maltodextrin and Guargum. The microcapsules were analyzed using Raman spectroscopy, total polyphenolcontent (TPC), and antioxidant capacity (Ax), and were subsequently subjectedto in vitro gastrointestinal digestion to assess their bioaccessibility.Raman spectra confirmed the formation of a maltodextrin-Guar-gum matrix withbroad glycosidic bands (480-1450 cm^-1) and CH-stretching at ≈2900 cm^-1,indicative of polymer-phenolic interactions. From de experimental design, theformulation containing 5% Guar gum at 100 ℃ reached the highest intestinal TPC(31.00 ± 0.30 mg GAE/100 g powder) and increased TPC bioaccessibility at the intestinalphase (283.28 ± 3.22%), evidencing efficient enzymatic release of boundphenolics. The greatest pre-digestion antioxidant capacity (19.56 ± 0.33% DPPHinhibition) corresponded to 7.5% GG at 104 ℃, while intestinal antioxidantrecovery peaked at 17.34 ± 0.14% (7.8% GG, 89.4 ℃). The optimal TPCbioaccessibility value obtained was 358.3%, under optimal spray-dryingconditions, consisting of 4% guar gum and an inlet temperature of 104 ℃. Overall,the synergy between NADES-based extraction and optimized spray-drying enabled astable, digestion-responsive encapsulation system that substantially enhancedphenolic retention and intestinal bioaccessibility, supporting its applicationas a sustainable strategy to valorize C. chinense leaves intoantioxidant-rich functional ingredients.

Keywords

Naturaldeep eutectic solvents / Spray drying / Microencapsulation / Capsicum chinense / Polyphenols / Antioxidant capacity / Bioaccessibility / In vitro digestion

Supplementary Materials

The supporting information associated with this article is available at https://www.sciepublish.com/article/pii/826: Figure S1: Calibration curve for the determination of total polyphenols in microencapsulated Capsicum chinense leaf extract experiments under in vitro digestion; Table S1: Total polyphenol content of microencapsulated Capsicum chinense extract experiments during in vitro digestion; Table S2: Antioxidant capacity of microencapsulated Capsicum chinense extract experiments during in vitro digestion; Table S3: Bioaccessibility results of total polyphenol content during in vitro digestion in the gastric and intestinal phases; Table S4: Regression Coefficients of the Response Surface for the Polyphenol Bioaccessibility during the Gastric Phase of Microencapsulated Habanero Pepper Leaf Extracts; Table S5: Results of the response surface ANOVA for the polyphenol bioaccessibility during the gastric phase of microencapsulated habanero pepper leaf extracts.

Author Contributions

Author Contributions: Conceptualization, I.M.R.-B.; Methodology, K.A.A.-B. and I.M.R.-B.; Software, M.O.R.-S.; Validation, K.A.A.-B., J.V.C.-R., M.O.R.-S. and I.M.R.-B.; Formal Analysis, K.A.A.-B. and J.V.C.-R.; Investigation, K.A.A.-B., J.V.C.-R. and I.M.R.-B.; Resources, I.M.R.-B.; Data Curation, K.A.A.-B.; Writing—Original Draft Preparation, K.A.A.-B.; Writing—Review & Editing, I.M.R.-B., M.O.R.-S. and J.V.C.-R.; Visualization, I.M.R.-B. and M.O.R.-S.; Supervision, I.M.R.-B. and J.V.C.-R.; Project Administration, I.M.R.-B.; Funding Acquisition, I.M.R.-B.

Ethics Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

The data presented in this study are available on request from the corresponding author.

Funding

Kevin Alejandro Avilés-Betanzos was supported by a SECIHTI scholarship (No. 661099).

Declaration of Competing Interest

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

References

[1] Alonso-Villegas R, González-Amaro RM, Figueroa-Hernández CY, Rodríguez-Buenfil IM. The Genus Capsicum: A Review of Bioactive Properties of Its Polyphenolic and Capsaicinoid Composition. Molecules 2023, 28, 4239. DOI: 10.3390/molecules28104239

[2] Herrera-Pool E, Patrón-Vázquez J, Ramos-Díaz A, Ayora-Talavera T, Pacheco N. Extraction and Identification of Phenolic Compounds in Roots and Leaves of Capsicum chinense by UPLC-PDA/MS. J. Bioeng. Biomed. Res. 2019, 3, 17-27. Available online: https://cmibq.org.mx/jbbr/images/doc/jbbr-vol-3-no-2/jbbr-vol-3-no-2-3.pdf?utm_source=chatgpt.com (accessed on 13 October 2025).

[3] DOF . Declaratoria General de Protección de la Denominación de Origen Chile Habanero de la Península de Yucatán; Diario Oficial de la Federación: Mexico City, Mexico, 2010. Available online: https://dof.gob.mx/nota_to_doc.php%3Fcodnota%3D5063632 (accessed on 26 October 2025).

[4] Herrera-Pool E, Ramos-Díaz AL, Lizardi-Jiménez MA, Pech-Cohuo S, Ayora-Talavera T, Cuevas-Bernardino JC, et al. Effect of Solvent Polarity on the Ultrasound Assisted Extraction and Antioxidant capacity of Phenolic Compounds from Habanero Pepper Leaves (Capsicum chinense) and Its Identification by UPLC-PDA-ESI-MS/MS. Ultrason. Sonochem. 2021, 76, 105658. DOI: 10.1016/j.ultsonch.2021.105658

[5] Olguín-Rojas JA, Vázquez-León LA, Palma M, Fernández-Ponce MT, Casas L, Fernández Barbero G, et al. Re-Valorization of Red Habanero Chili Pepper (Capsicum chinense Jacq.) Waste by Recovery of Bioactive Compounds: Effects of Different Extraction Processes. Agronomy 2024, 14, 660. DOI: 10.3390/agronomy14040660

[6] Dai Y, van Spronsen J, Witkamp GJ, Verpoorte R, Choi YH. Natural Deep Eutectic Solvents as a New Extraction Medium for Bioactive Compounds. Anal. Chim. Acta 2013, 766, 61-68. DOI: 10.1016/j.aca.2012.12.019

[7] Ruesgas-Ramón M, Figueroa-Espinoza MC, Durand E. Application of Deep Eutectic Solvents (DES) for Phenolic Compounds Extraction: Overview, Challenges, and Opportunities. J. Agric. Food Chem. 2017, 65, 3591-3601. DOI: 10.1021/acs.jafc.7b01054

[8] Socas-Rodríguez B, Torres-Cornejo MV, Álvarez-Rivera G, Mendiola JA. Deep Eutectic Solvents for the Extraction of Bioactive Compounds from Natural Sources and Agricultural By-Products. Appl. Sci. 2021, 11, 4897. DOI: 10.3390/app11114897

[9] Pavlić B, Mrkonjić Ž, Teslić N, Kljakić AC, Pojić M, Mandić A, et al. Natural Deep Eutectic Solvent (NADES) Extraction Improves Polyphenol Yield and Antioxidant capacity of Wild Thyme (Thymus serpyllum L.) Extracts. Molecules 2022, 27, 1508. DOI: 10.3390/molecules27051508

[10] Duarte H, Gomes V, Aliaño-González MJ, Faleiro L, Romano A, Medronho B. Ultrasound-Assisted Extraction of Polyphenols from Maritime Pine Residues with Deep Eutectic Solvents. Foods 2022, 11, 3754. DOI: 10.3390/foods11233754

[11] Ferreira LMDMC, Pereira RR, Carvalho-Guimarães FBD, Remígio MSDN, Barbosa WLR, Ribeiro-Costa RM, et al. Microencapsulation by Spray Drying and Antioxidant capacity of Phenolic Compounds from Tucuma Coproduct (Astrocaryum vulgare Mart.) Almonds. Polymers 2022, 14, 2905. DOI: 10.3390/polym14142905

[12] Ramakrishnan Y, Adzahan NM, Yusof YA, Muhammad K, Jailani F. Effect of Wall Materials on the Spray Drying Efficiency, Powder Properties and Stability of Bioactive Compounds in Tamarillo (Solanum betaceum) Juice Microencapsulation. Powder Technol. 2018, 328, 406-414. DOI: 10.1016/j.powtec.2017.12.018

[13] Macías-Cortés E, Gallegos-Infante JA, Rocha-Guzmán NE, Moreno-Jiménez MR, Villanueva-Fierro I, Ochoa-Martínez LA, et al. Spray Drying Conditions of Antioxidant and Anti-Inflammatory Polyphenols in Microcapsules of Ultrasound-Assisted Extract of Salvilla (Buddleja scordioides Kunth). ACS Food Sci. Technol. 2022, 2, 1574-1585. DOI: 10.1021/acsfoodscitech.2c00212

[14] Li Y, Tang B, Chen J, Lai P. Microencapsulation of plum (Prunus salicina Lindl.) phenolics by spray drying and storage stability. Food Sci. Technol. 2018, 38, 530-536. DOI: 10.1590/1678-457x.09817

[15] Bińkowska W, Szpicer A, Stelmasiak A, Wojtasik-Kalinowska I, Półtorak A. Microencapsulation of Polyphenols and Their Application in Food Technology. Appl. Sci. 2024, 14, 11954. DOI: 10.3390/app142411954

[16] Bernal-Millán MJ, Gutiérrez-Grijalva EP, Contreras-Angulo L, Muy-Rangel MD, López-Martínez LX, Heredia JB, et al. Spray-Dried Microencapsulation of Oregano (Lippia graveolens) Polyphenols with Maltodextrin Enhances Their Stability during In Vitro Digestion. J. Chem. 2022, 2022, 8740141. DOI: 10.1155/2022/8740141

[17] Avilés-Betanzos KA, Cauich-Rodríguez JV, González-Ávila M, Scampicchio M, Morozova K, Ramírez-Sucre MO, et al. Natural Deep Eutectic Solvent Optimization to Obtain an Extract Rich in Polyphenols from Capsicum chinense Leaves Using an Ultrasonic Probe. Processes 2023, 11, 1729. DOI: 10.3390/pr11061729

[18] Chel-Guerrero LD, Oney-Montalvo JE, Rodríguez-Buenfil IM. Phytochemical Characterization of By-Products of Habanero Pepper Grown in Two Different Types of Soils from Yucatán, Mexico. Plants 2021, 10, 779. DOI: 10.3390/plants10040779

[19] Avilés-Betanzos KA, Cauich-Rodríguez JV, Ramírez-Sucre MO, Rodríguez-Buenfil IM. Optimization of Spray Drying Conditions for a Capsicum chinense Leaf Extract Rich in Polyphenols Obtained by Ultrasonic Probe/NADES. ChemEngineering 2024, 8, 131. DOI: 10.3390/chemengineering8060131

[20] Pech-Pisté R, Pérez-Aranda C, Balam A, Vargas-Coronado R, Cauich-Rodríguez JV, Avilés F. Piezoimpedance of Carbon Nanotube Yarns Coupled with Raman Spectroscopy and Its Implementation for Sensing Polymerization Kinetics. Carbon 2023, 213, 118246. DOI: 10.1016/j.carbon.2023.118246

[21] Torres-Martínez BM, Vargas-Sánchez RD, Torrescano-Urrutia GR, González-Ávila M, Rodríguez-Carpena JG, Huerta-Leidenz N, et al. Use of Pleurotus ostreatus to Enhance the Oxidative Stability of Pork Patties during Storage and In Vitro Gastrointestinal Digestion. Foods 2022, 11, 4075. DOI: 10.3390/foods11244075

[22] Aguilera-Chávez SL, Gallardo-Velázquez T, Meza-Márquez OG, Osorio-Revilla G. Spray Drying and Spout-Fluid Bed Drying Microencapsulation of Mexican Plum Fruit (Spondias purpurea L.) Extract and Its Effect on In Vitro Gastrointestinal Bioaccessibility. Appl. Sci. 2022, 12, 2213. DOI: 10.3390/app12042213

[23] Chel-Guerrero LD, Castañeda-Corral G, López-Castillo M, Scampicchio M, Morozova K, Oney-Montalvo JE, et al. In Vivo Anti-Inflammatory Effect, Antioxidant capacity, and Polyphenolic Content of Extracts from Capsicum chinense By-Products. Molecules 2022, 27, 1323. DOI: 10.3390/molecules27041323

[24] Avilés-Betanzos KA, Oney-Montalvo JE, Cauich-Rodríguez JV, González-Ávila M, Scampicchio M, Morozova K, et al. Antioxidant Capacity, Vitamin C and Polyphenol Profile Evaluation of a Capsicum chinense By-Product Extract Obtained by Ultrasound Using Eutectic Solvent. Plants 2022, 11, 2060. DOI: 10.3390/plants11152060

[25] Yuen SN, Choi SM, Phillips DL, Ma CY. Raman and FTIR Spectroscopic Study of Carboxymethylated Non-Starch Polysaccharides. Food Chem. 2009, 114, 1091-1098. DOI: 10.1016/j.foodchem.2008.10.053

[26] Mir SA, Bosco SJD, Bashir M, Shah MA, Mir MM. Physicochemical and Structural Properties of Starches Isolated from Corn Cultivars Grown in Indian Temperate Climate. Int. J. Food Prop. 2017, 20, 821-832. DOI: 10.1080/10942912.2016.1184274

[27] Kizil R, Irudayaraj J, Seetharaman K. Characterization of Irradiated Starches by Using FT-Raman and FTIR Spectroscopy. J. Agric. Food Chem. 2002, 50, 3912-3918. DOI: 10.1021/jf011652p

[28] Różyło R, Szymańska-Chargot M, Zdunek A, Gawlik-Dziki U, Dziki D. Microencapsulated red powders from cornflower extract—Spectral (FT-IR and FT-Raman) and antioxidant characteristics. Molecules 2022, 27, 3094. DOI: 10.3390/molecules27103094

[29] Sakurai YCN, Pires IV, Ferreira NR, Moreira SGC, Silva LHMD, Rodrigues AMDC. Preparation and Characterization of Natural Deep Eutectic Solvents (NADESs): Application in the Extraction of Phenolic Compounds from Araza Pulp (Eugenia stipitata). Foods 2024, 13, 1983. DOI: 10.3390/foods13131983

[30] Milani A, del Zoppo M, Tommasini M, Zerbi G. The Effect of Intermolecular Dipole–Dipole Interaction on Raman Spectra of Polyconjugated Molecules: Density Functional Theory Simulations and Mathematical Models. J. Phys. Chem. B 2008, 112, 1619-1625. DOI: 10.1021/jp075763o

[31] Monge Neto AÁ, Bergamasco RDC, de Moraes FF, Medina Neto A, Peralta RM, Khodarahmi R. Development of a Technique for Psyllium Husk Mucilage Purification with Simultaneous Microencapsulation of Curcumin. PLoS ONE 2017, 12, e0182948. DOI: 10.1371/journal.pone.0182948

[32] Mudgil D, Barak S, Khatkar BS. Guar Gum: Processing, Properties and Food Applications—A Review. J. Food Sci. Technol. 2014, 51, 409-418. DOI: 10.1007/s13197-011-0522-x

[33] Ahmed HB, Emam HE. Environmentally Exploitable Biocide/Fluorescent Metal Marker Carbon Quantum Dots. RSC Adv. 2020, 10, 42916-42929. DOI: 10.1039/d0ra06383e

[34] Buljeta I, Pichler A, Šimunović J, Kopjar M. Polysaccharides as carriers of polyphenols: Comparison of freeze-drying and spray-drying as encapsulation techniques. Molecules 2022, 27, 5069. DOI: 10.3390/molecules27165069

[35] Drozłowska E, Starowicz M, Śmietana N, Krupa-Kozak U, Łopusiewicz Ł. Spray-Drying Impact the Physicochemical Properties and Formation of Maillard Reaction Products Contributing to Antioxidant capacity of Camelina Press Cake Extract. Antioxidants 2023, 12, 919. DOI: 10.3390/antiox12040919

[36] Samborska K. Powdered honey—Drying methods and parameters, types of carriers and drying aids, physicochemical properties and storage stability. Trends Food Sci. Technol. 2019, 88, 133-142. DOI: 10.1016/j.tifs.2019.03.019

[37] Hu H, Wang H, Zhang Y, Kan B, Ding Y, Huang J. Characterization of Genes in Guar Gum Biosynthesis Based on Quantitative RNA-Sequencing in Guar Bean (Cyamopsis tetragonoloba). Sci. Rep. 2019, 9, 10991. DOI: 10.1038/s41598-019-47518-5

[38] Xue H, Feng J, Tang Y, Wang X, Tang J, Cai X, et al. Research progress on the interaction of the polyphenol–protein–polysaccharide ternary systems. Chem. Biol. Technol. Agric. 2024, 11, 95. DOI: 10.1186/s40538-024-00632-7

[39] Karrar E, Mahdi AA, Sheth S, Mohamed Ahmed IA, Manzoor MF, Wei W, et al. Effect of maltodextrin combination with gum arabic and whey protein isolate on the microencapsulation of gurum seed oil using a spray-drying method. Int. J. Biol. Macromol. 2021, 171, 208-216. DOI: 10.1016/j.ijbiomac.2020.12.045

[40] Laureanti EJG, Paiva TS, de Matos Jorge LM, Jorge RMM. Microencapsulation of Bioactive Compound Extracts Using Maltodextrin and Gum Arabic by Spray and Freeze-Drying Techniques. Int. J. Biol. Macromol. 2023, 253, 126969. DOI: 10.1016/j.ijbiomac.2023.126969

[41] Čujić-Nikolić N, Stanisavljević N, Šavikin K, Kalaušević A, Nedović V, Bigović D, et al. Application of Gum Arabic in the Production of Spray-Dried Chokeberry Polyphenols, Microparticles Characterisation and In Vitro Digestion Method. Lek. Sirovine 2018, 38, 9-16. DOI: 10.5937/leksir1838009C

[42] Mahdi AA, Mohammed JK, Al-Ansi W, Ghaleb ADS, Al-Maqtari QA, Ma M, et al. Microencapsulation of Fingered Citron Extract with Gum Arabic, Modified Starch, Whey Protein and Maltodextrin Using Spray Drying. Int. J. Biol. Macromol. 2020, 152, 1125-1134. DOI: 10.1016/j.ijbiomac.2019.10.201

[43] Navarro-Flores MJ, Ventura-Canseco LMC, Meza-Gordillo R, Ayora-Talavera TDR, Abud-Archila M. Spray drying encapsulation of a native plant extract rich in phenolic compounds with combinations of maltodextrin and non-conventional wall materials. J. Food Sci. Technol. 2020, 57, 4111-4122. DOI: 10.1007/s13197-020-04447-w

[44] Robert P, Gorena T, Romero N, Sepúlveda E, Chávez J, Sáenz C. Encapsulation of Polyphenols and Anthocyanins from Pomegranate (Punica granatum) by Spray Drying. Int. J. Food Sci. Technol. 2010, 45, 1386-1394. DOI: 10.1111/j.1365-2621.2010.02270.x

[45] Šavikin K, Živković J, Alimpić Aradski A, Živković J, Menković N, Stević T, et al. Effect of Type and Concentration of Carrier Material on the Encapsulation of Pomegranate Peel Using Spray Drying Method. Foods 2021, 10, 1968. DOI: 10.3390/foods10091968

[46] Ricci A, Arboleda Mejia JA, Versari A, Chiarello E, Bordoni A, Parpinello GP. Microencapsulation of Polyphenolic Compounds Recovered from Red Wine Lees: Process Optimization and Nutraceutical Study. Food Bioprod. Process. 2022, 132, 1-12. DOI: 10.1016/J.FBP.2021.12.003

[47] Pieczykolan E, Kurek MA. Use of Guar Gum, Gum Arabic, Pectin, Beta-Glucan and Inulin for Microencapsulation of Anthocyanins from Chokeberry. Int. J. Biol. Macromol. 2019, 129, 665-671. DOI: 10.1016/j.ijbiomac.2019.02.073

[48] Cea-Pavez I, Manteca-Bautista D, Morillo-Gomar A, Quirantes-Piné R, Quiles JL. Influence of the Encapsulating Agent on the Bioaccessibility of Phenolic Compounds from Microencapsulated Propolis Extract during In Vitro Gastrointestinal Digestion. Foods 2024, 13, 425. DOI: 10.3390/foods13030425

[49] González Morales AN, López-Giraldo LJ, Sogamoso González E, Moscote Chinchilla Y. Evaluation of the Efficiency of Encapsulation and Bioaccessibility of Polyphenol Microcapsules from Cocoa Pod Husks Using Different Techniques and Encapsulating Agents. Processes 2025, 13, 3094. DOI: 10.3390/pr13103094