Synthesis of Chitosan Films Incorporated with Andrographis paniculata (Burm. F.) Nees Extract by Supercritical CO2 Technique and Their Biological Activities

Chau, Le Ngoc Quynh; Chau, Pham Trong Liem; Nhung, Tran Nguyen Cam; Oanh, Huynh Ngoc; Tien, Le Xuan; Hieu, Nguyen Huu

doi:10.48309/ajca.2026.572011.2035

Synthesis of Chitosan Films Incorporated with Andrographis paniculata (Burm. F.) Nees Extract by Supercritical CO₂ Technique and Their Biological Activities

Articles in Press

Document Type : Original Research Article

Authors

Le Ngoc Quynh Chau ¹^{, 2}^{, 3}

Pham Trong Liem Chau ¹^{, 2}^{, 3}

Tran Nguyen Cam Nhung ¹^{, 2}^{, 3}

Huynh Ngoc Oanh ¹^{, 2}^{, 3}

Le Xuan Tien ¹^{, 2}^{, 3}

Nguyen Huu Hieu ¹^{, 2}^{, 3}

¹ VNU-HCM, Key Laboratory of Chemical Engineering and Petroleum Processing (Key CEPP Lab), Ho Chi Minh City University of Technology (HCMUT), 268 Ly Thuong Kiet Street, Dien Hong Ward, Ho Chi Minh City, Vietnam

² Faculty of Chemical Engineering, Ho Chi Minh City University of Technology (HCMUT),268 Ly Thuong Kiet Street, Dien Hong Ward, Ho Chi Minh City, Vietnam

³ Vietnam National University Ho Chi Minh City, Linh Xuan Ward,Ho Chi Minh City, Vietnam

10.48309/ajca.2026.572011.2035

Abstract

In this study, Andrographis paniculata (Burm. F.) Nees extract was obtained using the supercritical carbon dioxide (CO₂) extraction method, a green and sustainable alternative to conventional solvent-based techniques. Chitosan (CTS) films, both with and without the A. paniculata extract (APE), were prepared using the solution-casting method. The effects of varying APE levels (1, 2, and 3 mL) incorporated into the CTS solution on the films' physical properties, such as color, light transmittance, moisture content, water solubility, and water vapor permeability, were systematically studied. The mechanical properties, including moisture content, swelling degree, and solubility, were also evaluated. Characterization of the synthesized films was conducted using advanced analytical techniques, including Fourier-transform infrared spectroscopy (FTIR), UV-visible absorption spectroscopy, scanning electron microscopy (SEM), and energy-dispersive X-ray spectroscopy (EDS). Additionally, the biological properties of the films were assessed, focusing on their antibacterial activity against Gram-positive Staphylococcus aureus and Gram-negative Pseudomonas aeruginosa, as well as their anti-inflammatory activity. Results revealed that CTS-2APE and CTS-3APE films exhibited significantly enhanced antibacterial activity compared to CTS alone, as evidenced by larger inhibition zones. Furthermore, the films containing APE demonstrated superior anti-inflammatory activity against egg albumin denaturation across various concentrations. These findings indicate that the incorporation of APE into CTS films enhances both their antibacterial and anti-inflammatory properties, making them promising candidates for biological activities.

Graphical Abstract

Keywords

Andrographis paniculata

Supercritical CO₂

Chitosan

Biological activities

Subjects

Applied Chemistry

OPEN ACCESS

©2026 The author(s). This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit: http://creativecommons.org/licenses/by/4.0/

PUBLISHER NOTE

Sami Publishing Company remains neutral concerning jurisdictional claims in published maps and institutional affiliations.

CURRENT PUBLISHER

Sami Publishing Company

[1] Mulla, M.Z., Ahmed, J., Vahora, A., Pathania, S. Effect of pectin incorporation on characteristics of chitosan based edible films. Journal of Food Measurement and Characterization, 2023, 17(6), 5569-5581.

[2] Zhao, J., Wang, Y., Li, J., Lei, H., Zhen, X., Gou, D., Liu, T. Preparation of chitosan/Enoki mushroom foot polysaccharide composite cling film and its application in blueberry preservation. International Journal of Biological Macromolecules, 2023, 246, 125567.

[3] Malik, Z., Parveen, R., Parveen, B., Zahiruddin, S., Khan, M.A., Khan, A., Husain, S.A. Anticancer potential of andrographolide from Andrographis paniculata (Burm. f.) Nees and its mechanisms of action. Journal of Ethnopharmacology, 2021, 272, 113936.

[4] Dey, S., Veerendra, G., Babu, P.A., Manoj, A.P., Nagarjuna, K. Degradation of plastics waste and its effects on biological ecosystems: A scientific analysis and comprehensive review. Biomedical Materials & Devices, 2024, 2(1), 70-112.

[5] Yuan, G., Lv, H., Yang, B., Chen, X., Sun, H. Physical properties, antioxidant and antimicrobial activity of chitosan films containing carvacrol and pomegranate peel extract. Molecules, 2015, 20(6), 11034-11045.

[6] Kaya, M., Khadem, S., Cakmak, Y.S., Mujtaba, M., Ilk, S., Akyuz, L., Deligöz, E. Antioxidative and antimicrobial edible chitosan films blended with stem, leaf and seed extracts of Pistacia terebinthus for active food packaging. RSC Advances, 2018, 8(8), 3941-3950.

[7] Carrera, C., Bengoechea, C., Carrillo, F., Calero, N. Effect of deacetylation degree and molecular weight on surface properties of chitosan obtained from biowastes. Food Hydrocolloids, 2023, 137, 108383.

[8] Pramanik, S., Aggarwal, A., Kadi, A., Alhomrani, M., Alamri, A.S., Alsanie, W.F., Bellucci, S. Chitosan alchemy: transforming tissue engineering and wound healing. RSC Advances, 2024, 14(27), 19219-19256.

[9] Kalariya, K.A., Gajbhiye, N.A., Meena, R.P., Saran, P.L., Minipara, D., Macwan, S., Geetha, K. Assessing suitability of Andrographis paniculata genotypes for rain-fed conditions in semi-arid climates. Information Processing in Agriculture, 2021, 8(2), 359-368.

[10] Shahbazi, Y. Characterization of nanocomposite films based on chitosan and carboxymethylcellulose containing Ziziphora clinopodioides essential oil and methanolic Ficus carica extract. Journal of Food Processing and Preservation, 2018, 42(2), e13444.

[11] Chau, T.P., Samdani, M.S., Kuriakose, L.L., Sindhu, R. Assessment of multi-biomedical efficiency of Andrographis paniculata shoot extracts through in-vitro analysis and major compound identification. Environmental Research, 2024, 242, 117779.

[12] Dinh, T.A., Le, Y.N., Pham, N.Q., Ton-That, P., Van-Xuan, T., Ho, T.G.T., Phuong, H.H.K. Fabrication of antimicrobial edible films from chitosan incorporated with guava leaf extract. Progress in Organic Coatings, 2023, 183, 107772.

[13] Wang, D., Shao, S., Wang, B., Guo, D., Tan, L., Chen, Q. Fabrication of chitosan/guar gum/polyvinyl alcohol films incorporated with polymethoxyflavone-rich citrus extracts: Postharvest shelf-life extension of strawberry fruits. Progress in Organic Coatings, 2024, 194, 108611.

[14] Tien, N.N.T., Nguyen, H.T., Le, N.L., Khoi, T.T., Richel, A. Biodegradable films from dragon fruit (Hylocereus polyrhizus) peel pectin and potato starches crosslinked with glutaraldehyde. Food Packaging and Shelf Life, 2023, 37, 101084.

[15] Dkhil, M.A., Zreiq, R., Hafiz, T.A., Mubaraki, M.A., Sulaiman, S., Algahtani, F., Al-Quraishy, S. Anthelmintic and antimicrobial activity of Indigofera oblongifolia leaf extracts. Saudi Journal of Biological Sciences, 2020, 27(2), 594-598.

[16] Rahman, L., Goswami, J., Choudhury, D. Assessment of physical and thermal behaviour of chitosan-based biocomposites reinforced with leaf and stem extract of Tectona grandis. Polymers and Polymer Composites, 2022, 30, 09673911221076305.

[17] Due, E., Rossen, K., Sorensen, L.T., Kliem, A., Karlsmark, T., Haedersdal, M. Effect of UV irradiation on cutaneous cicatrices: A randomized, controlled trial with clinical, skin reflectance, histological, immunohistochemical and biochemical evaluations. Acta Dermato-Venereologica, 2007, 87(1), 27-32.

[18] Özcan, G., Shenaq, S., Chahadeh, H., Spira, M. Ultraviolet-A induced delayed wound contraction and decreased collagen content in healing wounds and implant capsules. Plastic and Reconstructive Surgery, 1993, 92(3), 480-484.

[19] Nguyen, T.T., Dao, U.T.T., Bui, Q.P.T., Bach, G.L., Thuc, C.H., Thuc, H.H. Enhanced antimicrobial activities and physiochemical properties of edible film based on chitosan incorporated with Sonneratia caseolaris (L.) Engl. leaf extract. Progress in Organic Coatings, 2020, 140, 105487.

[20] Kalaycıoğlu, Z., Torlak, E., Akın-Evingür, G., Özen, İ., Erim, F.B. Antimicrobial and physical properties of chitosan films incorporated with turmeric extract. International Journal of Biological Macromolecules, 2017, 101, 882-888.

[21] Sánchez-González, L., Vargas, M., González-Martínez, C., Chiralt, A., Cháfer, M. Characterization of edible films based on hydroxypropylmethylcellulose and tea tree essential oil. Food Hydrocolloids, 2009, 23(8), 2102-2109.

[22] Siripatrawan, U., Vitchayakitti, W. Improving functional properties of chitosan films as active food packaging by incorporating with propolis. Food Hydrocolloids, 2016, 61, 695-702.

[23] Obagi, Z.; Damiani, G.; Grada, A.; Falanga, V. Principles of wound dressings: A review. Surg. Technol. Int, 2019, 35(5), 50-57.

[24] Cutting, K.F. Wound exudate: composition and functions. British Journal of Community Nursing, 2003, 8(Sup3), S4-S9.

[25] Pereda, M., Ponce, A., Marcovich, N., Ruseckaite, R., Martucci, J. Chitosan-gelatin composites and bi-layer films with potential antimicrobial activity. Food Hydrocolloids, 2011, 25(5), 1372-1381.

[26] Ojagh, S.M., Rezaei, M., Razavi, S.H., Hosseini, S.M.H. Development and evaluation of a novel biodegradable film made from chitosan and cinnamon essential oil with low affinity toward water. Food Chemistry, 2010, 122(1), 161-166.

[27] Souza, V.G.L., Fernando, A.L., Pires, J.R.A., Rodrigues, P.F., Lopes, A.A., Fernandes, F.M.B. Physical properties of chitosan films incorporated with natural antioxidants. Industrial Crops and Products, 2017, 107, 565-572.

[28] Aleem, A.R., Shahzadi, L., Tehseen, S., Alvi, F., Chaudhry, A.A., ur Rehman, I., Yar, M. Amino acids loaded chitosan/collagen based new membranes stimulate angiogenesis in chorioallantoic membrane assay. International Journal of Biological Macromolecules, 2019, 140, 401-406.

[29] Rihayat, T., Hadi, A.E., Aidy, N., Safitri, A., Siregar, J.P., Cionita, T., Fitriyana, D.F. Biodegradation of polylactic acid-based bio composites reinforced with chitosan and essential oils as anti-microbial material for food packaging. Polymers, 2021, 13(22), 4019.

[30] Yong, H., Wang, X., Bai, R., Miao, Z., Zhang, X., Liu, J. Development of antioxidant and intelligent pH-sensing packaging films by incorporating purple-fleshed sweet potato extract into chitosan matrix. Food Hydrocolloids, 2019, 90, 216-224.

[31] Liang, Y., He, J., Guo, B. Functional hydrogels as wound dressing to enhance wound healing. ACS Nano, 2021, 15(8), 12687-12722.

[32] Pagliarulo, C., De Vito, V., Picariello, G., Colicchio, R., Pastore, G., Salvatore, P., Volpe, M.G. Inhibitory effect of pomegranate (Punica granatum L.) polyphenol extracts on the bacterial growth and survival of clinical isolates of pathogenic Staphylococcus aureus and Escherichia coli. Food Chemistry, 2016, 190, 824-831.

[33] Li, J., Zhuang, S. Antibacterial activity of chitosan and its derivatives and their interaction mechanism with bacteria: Current state and perspectives. European Polymer Journal, 2020, 138, 109984.