Obtención de colorante en polvo liofilizado a partir de la flor de Hibiscus rosa-sinensis J. Food Sci. Gastron. (January - June 2026) 4(1): 10-16 https://doi.org/10.5281/zenodo.18294106 ISSN 3073-1283 ORIGINAL ARTICLE Obtaining freeze-dried powdered dye from Hibiscus rosa-sinensis flowers  José A. Arencibia jose.arencibia1995@gmail.com 1 Instituto de Farmacia y Alimentos, Universidad de la Habana, Cuba. Received: 27 September 2025 / Accepted: 14 December 2025 / Published online: 23 January 2026 © The Author(s) 2026 José A. Arencibia 1,2 · Dairon Iglesias 1 · Ariel A. Vergel 2 · Alicia Casariego 1 Abstract Currently, there is growing interest in bioactive plant compounds, such as anthocyanins, pigments wide- ly distributed in fruits and ﬂowers, which not only provide color but also antioxidant properties with preventive eﬀects against coronary heart disease and certain types of cancer. The objective of this study was to develop a natural powdered dye by freeze-drying Hibiscus rosa-sinensis ﬂowers, contrib- uting to the agri-food heritage through the agro-industrial use of plant species. The collected ﬂowers were subjected to hydroalcoholic extraction, concentration, and freeze-dry- ing according to the proposed experimental design. The op- timal formulation was obtained with 8.2% maltodextrin, 5% starch, and 1.863% xanthan gum. The resulting dye achieved an encapsulation eﬃciency of 83.67%, moisture content of 9.48%, hygroscopicity of 17.98%, and solubility of 90.23%, in accordance with the theoretical values determined by nu- merical optimization. These ﬁndings demonstrate the poten- tial for innovation in the natural colorant industry, aligning the valorization of plant resources with the principles of sus- tainable agri-food heritage. Keywords Hibiscus rosa-sienensis L., natural coloring, freeze-drying, anthocyanins, optimization. Resumen Actualmente, existe un creciente interés por los compuestos bioactivos vegetales, como las antocianinas, pigmentos ampliamente distribuidos en frutas y ﬂores, que no solo aportan color sino también propiedades antioxidan- tes con efectos preventivos frente a enfermedades corona- rias y ciertos tipos de cáncer. El objetivo de este estudio fue desarrollar un colorante natural en polvo mediante lioﬁliza- ción de ﬂores de Hibiscus rosa-sinensis, contribuyendo al patrimonio agroalimentario a través del aprovechamiento agroindustrial de especies vegetales. Las ﬂores recolecta- das fueron sometidas a extracción hidroalcohólica, con- centración y lioﬁlización conforme al diseño experimental propuesto. La formulación óptima se obtuvo con 8,2 % de maltodextrina, 5 % de almidón y 1,863 % de goma xantana. El colorante resultante alcanzó una eﬁciencia de encapsula- ción del 83,67 %, humedad del 9,48 %, higroscopicidad del 17,98 % y solubilidad del 90,23 %, en concordancia con los valores teóricos determinados mediante optimización numé- rica. Estos hallazgos evidencian el potencial de innovar en la industria de colorantes naturales, alineando la valorización de recursos vegetales con los principios del patrimonio agro- alimentario sostenible. Palabras clave Hibiscus rosa-sienensis L., colorante natu- ral, lioﬁlización, antocianinas, optimización. 1 Instituto de Farmacia y Alimentos, Universidad de la Habana, Cuba. 2 Facultad Tecnológica, Universidad de Santiago de Chile, Chile. 3 Departamento de Investigación y Posgrado en Alimentos, Universidad de Sonora, México. How to cite Arencibia, J. A., Iglesias, D., Vergel, A. A., & Casariego, A. (2026). Obtaining freeze-dried powdered dye from Hibiscus rosa-sinensis ﬂowers. Journal of Food Science and Gastronomy, 4(1), 10-16. https://doi.org/10.5281/zenodo.18294106

J. Food Sci. Gastron. (January - June 2026) 4(1): 10-16 11 Introduction The demand for natural food colorants has increased signiﬁcantly, driven by consumer preference for clean-label products and growing toxicological concerns associated with synthetic colorants (Gaibor et al., 2022). The ban and regulatory reevaluation of several of these additives by agencies such as the FDA have led to a boom in the search for safe natural alternatives. In this context, anthocyanins, water-soluble pigments responsible for red, blue, and purple hues, are emerging as highly promising candidates (Mejía et al., 2020). The ﬂowers of Hibiscus rosa-sinensis L. are a signiﬁcant source of anthocyanins, predominantly cyanidin- 3-sophoroside. Previous studies have demonstrated the feasibility of their extraction and application as colorants and in smart packaging systems, including solid-liquid kinetics analysis with ethanol-water solvents and evaluation of their antioxidant capacity (Pérez-Orozco et al., 2020; Mejía et al., 2023). However, the application of anthocyanins is limited by their marked instability in the face of environmental factors such as light, pH, oxygen, and temperature (Mohammadalinejhad & Kurek, 2021). In this context, microencapsulation has established itself as an eﬀective strategy for protecting the bioactive compound by incorporating it into wall matrices, increasing its stability and shelf life during storage (de Moura et al., 2018). Although this technique has been applied to anthocyanins from various sources, its optimization for H. rosa-sinensis remains poorly documented, constituting a knowledge gap that the present study addresses. In this context, the present study aims to optimize a mi- croencapsulation process to obtain powdered dye from H. rosa-sinensis ﬂowers. The research is distinguished by us- ing food-grade solvents and the selection of wall materials aimed at maximizing encapsulation eﬃciency. The resulting product combines color stability and antioxidant functional- ity, providing a relevant techno-functional beneﬁt for food preservation and positioning itself as a diﬀerentiated alterna- tive to synthetic dyes. Methodology HRS ﬂowers were collected and selected for uniformity in size, color, and absence of visible morphological altera- tions. Petals and pistils were separated and dried in an oven (YLD-6000, AISET, China) at 40 °C for 72 hours. They were then ground with a handmade grinder, sieved (0.5 mm), and stored in amber bottles with ground glass stoppers in a desic- cator until use (Arencibia et al., 2023). The extract was obtained by maceration for 24 hours at 60 °C, with 50% ethanol (v/v) acidiﬁed with 0.5% citric acid, at a solid/liquid ratio of 1 g/20 ml (Arencibia et al., 2023). The mixture was then ﬁltered using a vacuum pump (UL- VAC DTC-21), and the ﬁltrate was concentrated to 7% total solids by rotary evaporation (RV 10 Basic, IKA, Argentina) at 40 °C and 85 s -1 . To 35 ml of the concentrated extract (7% total solids), diﬀerent combinations of between 5 and 9% maltodextrin (MX) DE 20 and modiﬁed starch (MS) were added, toge- ther with xanthan gum (XG) at concentrations between 0 and 2%, according to the experimental design in Table 1. The formulations were homogenized in a magnetic stirrer (ASIN MS-500) at 750 rpm, adjusting the total solids between 20 and 25% according to the corresponding run. Finally, the samples were subjected to freeze-drying (Telstar Lioalfa-6 Lyophyliser), operating at -50 °C and 10 -2 Pa for 36 hours. The encapsulation eﬃciency (EE) was determined by quantifying total and surface anthocyanins according to Lee et al. (2005). Absorbances at 510 and 700 nm were measured in buﬀer solutions at diﬀerent pH values using spectrophoto- metry (Rayleigh UV-1601, Beijing), applying the equations proposed by Robert et al. (2010). Moisture content (Hu) was determined by drying in an oven at 105 °C (YLD-6000 AISET, China), calculated by the diﬀerence in weight before and after drying (da Rosa et al., 2019). Hygroscopicity (Hi) was determined by exposing 1 g of dye to a controlled atmosphere with saturated sodium chloride solution (75.29% RH at 25 °C) and was expressed as g of water/100 g of dry solids (Tonon et al., 2008). Solu- bility (Sol) was evaluated by quantifying the percentage of powder dissolved in distilled water through stirring, centrifu- gation, and drying of the recovered solids. It was calculated by comparing the initial weight with that of the dry solids (Cano-Chauca et al., 2005). The characterization of the optimal extract replicated the methodologies described above. The procedures were re- plicated three times to ensure accuracy. Statistical analysis was performed using simple ANOVA in Statistics (version 7, 2004, StatSoft Inc., Tulsa, USA), applying Duncan’s test (p ≤ 0.05) for comparison of means. Results and discussion The results shown in Table 1 indicate that, among the formulations evaluated, runs 3, 4, 6, and 8 presented high EE (≥ 84%) and Sol above 86%, accompanied by low levels of Hy (16.0–17.3%), suggesting the formation of denser and more cohesive matrices. Run 3 achieved the highest EE (85.7%) and one of the lowest Hy (16%), supporting the hypothesis that a balanced proportion of the three components promotes the formation of a continuous and highly soluble encapsulating matrix. Likewise, run 4 recorded the highest Sol value (88.6%). In contrast, the formulations corresponding to runs 5 and 7 showed lower

J. Food Sci. Gastron. (January - June 2026) 4(1): 10-16 12 EE values of 61.0 and 66.4%, respectively, accompanied by high Hy (up to 21.9%) and lower Sol levels, which can be attributed to the low concentration of encapsulating agents and the absence of XG in the mixture. Table 1. Experimental design and response variables Run A (%) B (%) C (%) EE (%) Hu (%) Hy (%) Sol (%) Mean (Standard deviation) Mean (Standard deviation) Mean (Standard deviation) Mean (Standard deviation) 1 7.2 6.8 0.043 71.5 (0.6) c 8.76 (0.02) bc 19.4 (0.2) ef 84.7 (0.8) ab 2 9 9 0 70.8 (0.7) c 9.8 (0.6) de 16.0 (1) a 83.9 (2.7) a 3 9 7.5 2 85.7 (0.5) e 10.1 (0.8) e 16.0 (0.9) a 87.0 (1.3) bc 4 7.2 8.94 1.1 84.2 (1) e 9.5 (0.2) de 16.8 (0.2) ab 88.6 (1.9) d 5 5 5 0 61.0 (3) a 7.91 (0.11) a 21.9 (0.5) g 85.4 (0.2) abc 6 9 5 0.74 84.7 (0.2) e 9.31 (0.41) cd 17.7 (0.4) c 85.2 (0.5) abc 7 5 9 0 66.4 (0.6) b 8.42 (0.15) ab 18.2 (0.4) cd 85.3 (0.8) abc 8 7.2 8.94 1.1 84.3 (0.8) e 9.5 (0.6) de 17.3 (0.3) bc 86.5 (0.4) abc 9 5.08 6.8 1.1 77.1 (0.6) d 8.39 (0.06) ab 19.85 (0.08) f 86.8 (1.7) abc 10 6.46 5 2 76.5 (0.4) d 9.2 (0.2) cd 18.8 (0.3) de 87.9 (1.8) cd 11 5.08 6.8 1.1 76.3 (0.2) d 8.39 (0.01) ab 19.0 (0.2) def 86.4 (1.7) abc 12 5 9 2 78.0 (3) d 9.22 (0.08) cd 18 (1) bc 87.9 (1.7) c 13 7.2 6.8 0.043 71.3 (0.3) c 8.7 (0.1) bc 19.3 (0.3) ef 84.7 (0.4) ab A: maltodextrin, B: modiﬁed starch, C: xanthan gum, EE: encapsulation eﬃciency, Hu: humidity, Hy: hygroscopicity, Sol: solubility. Diﬀerent letters indicate signiﬁcant diﬀerences (p ≤ 0.05). Table 2 presents the results of the analysis of variance (ANOVA) of the coded models used to describe the eﬀect of the independent variables on EE, Hu, Hi, and Sol of the encapsulated dye. Terms with ≤ 0.05 were considered signiﬁcant. Table 2. Analysis of variance of coded models for encapsulation eﬃciency, moisture content, hygroscopicity, and dye solubility Parameter p value EE Hu Hy Sol Model < 0.0001 < 0.0001 < 0.0001 0.0023 A < 0.0001 < 0.0001 < 0.0001 0.1238 B < 0.0001 < 0.0001 < 0.0001 0.4711 C < 0.0001 < 0.0001 0.0021 0.0004 AB < 0.0001 0.0187 - - AC 0.0002 - - - BC - - - - A 2 < 0.0001 - - - B 2 < 0.0001 0.0004 - - C 2 < 0.0001 0.0076 - - Lack of ﬁt 0.7671 0.2733 0.3081 0.6690 A: maltodextrin, B: modiﬁed starch, C: xanthan gum, EE: encapsulation eﬃciency, Hu: humidity, Hy: hygroscopicity, Sol: solubility. The models adjusted for each response variable were statistically signiﬁcant, indicating that at least one term contributes to explaining the observed variability. The lack of adjustment was not signiﬁcant, supporting the validity and representativeness of the models. In the EE model, all main terms (A, B, C), interactions (AB, AC), and quadratic terms (A², B², C²) were signiﬁcant, showing linear, nonlinear, and interactive eﬀects. For Hu, A, B, C, the AB interaction, and the quadratic terms B² and C² were signiﬁcant, reﬂecting combined and curvilinear eﬀects. In Hy, A and B were signiﬁcant, while C showed a relevant eﬀect without interaction or quadratic terms. In Sol, only C was signiﬁcant, indicating XG as a determining component. Although non-signiﬁcant terms were eliminated, Gutiérrez & De la Vara (2012) recommend preserving the hierarchy for a robust statistical interpretation. The R² values were high (78.5–99.9%). The inﬂuence of A, B, and C on the response variables (EE, Hu, Hy, and Sol) was evaluated using response surface graphs, which show the average behavior according to the adjusted model (Gutiérrez & De la Vara, 2012). The joint analysis of Figures 1-4 reveals the diﬀerential role of XG (C), MX (A), and MS (B) on the response variables evaluated. In Figure 1, XG has a positive and signiﬁcant eﬀect on EE, especially from 0.74% onwards, reaching values close to 90% in the optimal zone. In the absence of XG, EE depended more on A and B, but its combination with C generated synergistic interactions that stabilize the response (EE). In Figure 2, XG does not have a signiﬁcant linear eﬀect on Hu at levels below 2%, maintaining values between 7.5 and 10.5%. However, at 2%, a moderate increase is observed associated with its water retention capacity. MX has a reducing eﬀect (Nik Abd Rahman et al., 2024), while MS slightly increases it at high concentrations due to its hydrophilic nature. In Figure 3, Hy is dominated by A and B, with a signiﬁcant positive eﬀect of MX and a negative eﬀect of MS, like the

J. Food Sci. Gastron. (January - June 2026) 4(1): 10-16 13 Figure 1. Inﬂuence of xanthan gum on EE. a) 0%; b) 0.0434446%; c) 0.74%; d) 1.1% y e) 2%. Figure 2. Inﬂuence of xanthan gum on humidity. a) 0%; b) 0.0434446%; c) 0.74%; d) 1.1% y e) 2%. study by Tonon et al. (2008). XG acts as a modulator (C and C², p < 0.01), smoothing the gradients and stabilizing Hyat intermediate and high levels, which could be related to a denser matrix that limits Hu absorption. In Figure 4, XG shows a positive and signiﬁcant eﬀect on Sol (C and C², p < 0.01), with the maximum at 2%. Its high solubility and rapid hydration facilitate dye dispersion. Meanwhile, A and B show no signiﬁcant eﬀects or interactions with C, indicating that Sol is mainly determined by XG content. Overall, XG appears to be the main control factor in EE and Sol, a modulator of Hy, and to have a limited eﬀect on Hu, while MX and MS play opposite and complementary roles in Hy and Hu, helping to optimize the encapsulating matrix. Optimization of response variables After adjusting and validating the model, the response

J. Food Sci. Gastron. (January - June 2026) 4(1): 10-16 14 surface was explored to deﬁne optimal conditions or directions for future experimentation (Gutiérrez & De la Vara, 2012). In this study, the validated model made it possible to establish the ideal conditions for obtaining a natural powdered dye from HRS ﬂowers, simultaneously considering all process variables. Table 3 shows the constraints applied to optimize EE, Hu, Hy, and Sol. Fourteen feasible solutions were obtained, with solution 1 being selected for its greater statistical convenience: 8.2% MX, 5% MS, and 1.863% XG. Under these conditions, 85.28% EE, 9.57% Hu, 18.00% Hy, and 87.11% Sol were predicted, with a convenience of 0.701. The dye obtained under optimal conditions showed an EE of 83.67% (Table 4), which is lower than that estimated by numerical optimization. However, the relative percentage error was 1.89%, indicating high precision and experimental reliability. A lower percentage of Hu was obtained than estimated by the model. XG, with limited environmental absorption but high internal water retention, probably modulated the free water in the formulation, reducing the Figure 3. Inﬂuence of xanthan gum on hygroscopicity. a) 0%; b) 0.0434446%; c) 0.74%; d) 1.1% y e) 2.2%. Figure 4. Inﬂuence of xanthan gum on solubility. a) 0%; b) 0.0434446%; c) 0.74%; d) 1.1% y e) 2%.

J. Food Sci. Gastron. (January - June 2026) 4(1): 10-16 15 ﬁnal Hu of the product to 9.48% (Patel et al., 2020; Berninger et al., 2021). Table 3. Constraints for optimizing the dye production process Parameter(%) Minimum limit Maximum limit Criterion Maltodextrin 5 9 Maximize Modiﬁed starch 5 9 Minimize Xanthan gum 0 2 Maximize Encapsulation eﬃciency 61.0 85.7 Maximize Humidity 7.9 10.1 Minimize Hygroscopicity 16.0 22.0 Minimize Solubility 83.9 88.6 Maximize Table 4. Physical and chemical indicators of the optimal dye obtained Parameter Mean Standar deviation Encapsulation eﬃciency (%) 83.67 0.09 Humidity (%) 9.48 0.02 Hygroscopicity (%) 17.98 0.14 Solubility (%) 90.23 0.36 The optimal formulation yielded a Hy of 17.98%, a moderate value suggesting that XG stabilized the product against RH variations without increasing its absorption. Unlike other hygroscopic polysaccharides, XG is not particularly hygroscopic (Berninger et al., 2021), regulating internal Hu and water-powder balance. Due to its high solubility and ability to form viscous solutions, XG improves the dispersion of powders in water, especially in low proportions (Nsengiyumva & Alexandridis, 2022). In the optimal formulation, a Sol of 90.23% was obtained, higher than estimated, showing that XG did not limit but rather favored the dissolution of the active components. Conclusions Xanthan gum proved to be a key component in contro- lling the physicochemical properties of the powdered dye obtained. Its use in combination with other encapsulants op- timized encapsulation eﬃciency, controlled internal moistu- re, stabilized hygroscopicity, and promoted high solubility. These eﬀects are directly related to its molecular structure and rheological behavior, which provide stability without compromising product functionality. References Arencibia, J. A., García, C. L., Morgan, A. D. L., Salas-Oli- vet, E., García-Beltrán, J. A., & Casanova, R. M. (2023). Optimización del proceso de extracción de an- tocianinas y polifenoles a partir de las ﬂores de Hibis- cus rosa-sinensis L. Ciencia y Tecnología de Alimentos, 33(3), 1-11. https://revcitecal.iiia.edu.cu/revista/index. php/RCTA/es/article/view/690 Berninger, T., Dietz, N., & González López, Ó. (2021). Water‐soluble polymers in agriculture: xanthan gum as eco‐friendly alternative to synthetics. Microbi- al Biotechnology, 14(5), 1881-1896. https://doi. org/10.1111/1751-7915.13867 Cano-Chauca, M., Stringheta, P. C., Ramos, A. M., & Cal- Vidal, J. (2005). Eﬀect of the carriers on the microstruc- ture of mango powder obtained by spray drying and its functional characterization. Innovative Food Science & Emerging Technologies, 6(4), 420-428. https://doi. org/10.1016/j.ifset.2005.05.003 da Rosa, J. R., Nunes, G. L., Motta, M. H., Fortes, J. P., Weis, G. C. C., Hecktheuer, L. H. R., Muller, E. I., Ragag- nin, C., & da Rosa, C. S. (2019). Microencapsulation of anthocyanin compounds extracted from blueberry (Vaccinium spp.) by spray drying: Characterization, sta- bility and simulated gastrointestinal conditions. Food hydrocolloids, 89, 742-748. https://doi.org/10.1016/j. foodhyd.2018.11.042 de Moura, J. S., Berling, C., Germer, S. P. M., Alvim, I. D., & Hubinger, M. D. (2018). Encapsulating anthocyanins from Hibiscus sabdariﬀa L. calyces by ionic gelation: Pigment stability during storage of microparticles. Food Chemistry, 241, 317–327. https://doi.org/10.1016/j. foodchem.2017.08.095 Gaibor, F. M., Rodríguez, D., García, M. A., Peraza, C. M., Vidal, D., Nogueira, A., & Casariego, A. (2022). De- velopment of a food colorant from Syzygium cumini L. (Skeels) by spray drying. Journal of Food Science and Technology, 59, 4045-4055. https://doi.org/10.1007/ s13197-022-05454-9 Gutiérrez, H., & De la Vara, R. (2012). Análisis y diseño de experimentos (3.ª ed.). México, D.F.: McGraw-Hill. https://bibliotecadigital.uce.edu.ec/s/L-D/item/1166 Lee, J., Durst, R. W., Wrolstad, R. E., Eisele, T., Giusti, M. M., Hach, J., Hofsommer, H., Koswig, S., Krueger, D. A., Kupina, S., Martin, S., Martinsen, S. K., Miller, T. C., Paquette, F., Ryabkova, A., Skrede, G., Trenn, U., & Wightman, J. D. (2005). Determination of to- tal monomeric anthocyanin pigment content of fruit juices, beverages, natural colorants, and wines by the pH diﬀerential method: collaborative study. Journal of AOAC international, 88(5), 1269-1278. https://doi. org/10.1093/jaoac/88.5.1269

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