Immobilization of Chaga extract in alginate beads for modified release: simplicity meets efficiency

Main Article Content

Predrag Petrović
Katarina Ivanović
Charly Octrue
Mića Tumara
Aleksandra Jovanović
Jovana Vunduk
Miomir Nikšić
Rada Pjanović
Branko Bugarski
Anita Klaus


Chaga (Inonotus obliquus) is a parasitic fungus which has been used in traditional medicine in Russia and other northern European countries in treatment of cancer, gastrointestinal, liver and other diseases. It has been a subject of intensive research recently, confirming many of its health-beneficial effects. In order to obtain a product that would allow modified and prolonged release of the Chaga’s active metabolites, hot water Chaga extract was encapsulated using sodium alginate. The extract, which was predominantly made of carbohydrates (57%), also contained relatively large amount of antioxidants/phenolic compounds (130 mg gallic acid equivalents per g of dry extract) and exhibited pronounced radical scavenging activity. It showed significant antibacterial activity as well, inhibiting growth of tested bacterial strains at concentrations of 1.25-20 mg/mL. Encapsulation efficiency was almost 80%, and the extract-alginate system showed pH-dependant extract release; there was almost no release at pH 1.75 (gastric pH), and the release gradually increased with the rise of the pH level, reaching maximum of about 43% of released extract at pH=8.5 after 90 min. Such product could be used as a dietary supplement, adjuvant in therapy of gastrointestinal diseases or as a food additive.

Article Details

Chemical Engineering - Pharmaceutical Engineering
Author Biography

Rada Pjanović, Faculty of Technology and Metallurgy, University of Belgrade, Belgrade

Faculty of Technology and Metallurgy, University of Belgrade, Belgrade


Grover A, Joshi, A. An Overview of Chronic Disease Models: A Systematic Literature Review. Glob J Health Sci. 2014; 7(2): 210-227.

Zaman SB, Hussain MA, Nye R, Mehta V, Mamun KT, Hossain N. A Review on Antibiotic Resistance: Alarm Bells are Ringing. Cureus. 2017; 9(6): e1403

Bills GF, Gloer JB. Biologically Active Secondary Metabolites from the Fungi. Microbiol. Spectr. 2016; 4(6): 1-32.

Padmavathi M. Chronic Disease Management with Nutraceuticals. Int J Pharm Sci Invent. 2013; 2(4): 1-11

Patel S. Chaga (Inonotus Obliquus) Mushroom: Nutraceutical Assesement Based on Latest Findings. In: Patel S, ed. Emerging Bioresources with Nutraceutical and Pharmaceutical Prospects. Basingstoke, England: Springer; 2015: 115-126

Lemieszek MK, Langner E, Kaczor J, Kandefer-Szerszen M, Sanecka B, Mazurkiewicz W, & Rzeski W. Anticancer Effects of Fraction Isolated from Fruiting Bodies of Chaga Medicinal Mushroom, Inonotus obliquus (Pers.:Fr.) Pilát (Aphyllophoromycetideae): In Vitro Studies. Int. J. Med. Mushrooms. 2011; 13(2): 131-143.

Shashkina MY, Shashkin PN, Sergeev, AV. Chemical and medicobiological properties of chaga (review). Pharm Chem J. 2006; 40(10): 560-568.

Lee J, Hyun C. Insulin-Sensitizing and Beneficial Lipid-Metabolic Effects of the Water-Soluble Melanin Complex Extracted from Inonotus obliquus. Phytother. Res. 2014; 28(9): 1320-1328.

Liu C, Zhao C, Pan H, Kang J, Yu X, Wang, H, Li B, Xie Y, Chen, R. Chemical Constituents from Inonotus obliquus and Their Biological Activities. J. Nat. Prod. 2013; 77(1): 35-41.

Hwang BS, Lee I, Yun B. Phenolic compounds from the fungus Inonotus obliquus and their antioxidant properties. J Antibiot. 2015; 69(2): 108-110.

Rijo P, Matias D, Fernandes A, Simões M, Nicolai M, Reis C. Antimicrobial Plant Extracts Encapsulated into Polymeric Beads for Potential Application on the Skin. Polymers. 2014; 6(2): 479-490.

Pasukamonset P, Kwon O, Adisakwattan S. Alginate-based encapsulation of polyphenols from Clitoria ternatea petal flower extract enhances stability and biological activity under simulated gastrointestinal conditions. Food Hydrocolloid. 2016; 61: 772–779.

Fang Z, Bhandari B. Encapsulation of polyphenols - a review. Trends Food Sci. Technol. 2010; 21: 510–523.

Amatrejo S, Hr T. Tinospora extract (Tinospora crispa (L.) Miers.) encapsulation with sodium alginate isolated from brown algae (Sargassum ilicifolium). Asian J Pharm Clin Res. 2017; 10(11): 267-269.

Chan E, Yim Z, Phan S, Mansa RF, Ravindra P. Encapsulation of herbal aqueous extract through absorption with ca-alginate hydrogel beads. Food Bioprod Process. 2010; 88(2-3): 195-201.

Balanč B, Kalušević A, Drvenica I, Coelho MT, Đorđević V, D Alves V, Sousa I, Moldao-Martins M, Rakić V, Nedović V, Bugarski B. Calcium-alginate-inulin microbeads as carriers for aqueous carqueja extract. J. Food Sci. 2016; 81: E65–E75.

Jovanovic A, Balanč B, Djordjević V, Ota A, Skrt M, Savikin K, Bugarski B, Nedović V, Ulrih Poklar N. Effect of gentisic acid on the structural-functional properties of liposomes incorporating β-sitosterol. Colloid Surface B. 2019;

Đorđević V, Balanč B, Belščak-Cvitanović A, Lević S, Trifković K, Kalušević A, Kostić I, Komes D, Bugarski B, Nedović V. Trends in encapsulation technologies for delivery of food bioactive compounds. Food Eng Rev. 2015; 7: 452–490.

Gorbunova N, Bannikova A, Evteev A, Evdokimov I, Kasapis S. Alginate-based encapsulation of extracts from beta Vulgaris cv. beet greens: Stability and controlled release under simulated gastrointestinal conditions. LWT. 2018; 93: 442-449.

Desai KG, Jin Park H. Recent Developments in Microencapsulation of Food Ingredients. Dry Technol. 2005; 23(7): 1361-1394.

DuBois M, Gilles KA, Hamilton JK, Rebers PA, Smith F. Colorimetric Method for Determination of Sugars and Related Substances. Anal Chem. 1956; 28(3): 350-356.

Bradford M. A Rapid and Sensitive Method for the Quantitation of Microgram Quantities of Protein Utilizing the Principle of Protein-Dye Binding. Anal Biochem. 1976; 72(1-2): 248-254.

Jovanović A, Đorđević V, Zdunić G, Šavikin К, Pljevljakušić D, Bugarski B. Ultrasound-assisted extraction of polyphenols from Thymus serpyllum and its antioxidant activity. Hem Ind. 2016; 70: 391–398.

Re R, Pellegrini N, Proteggente A, Pannala A, Yang M, Rice-Evans C. Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radic Biol Med. 1999; 26(9-10): 1231-1237.

Petrović P, Vunduk J, Klaus A, Carević M, Petković M, Vuković N, Cvetković A, Žižak Ž, Bugarski B. (2019). From mycelium to spores: A whole circle of biological potency of mosaic puffball. ‎S. Afr. J. Bot. 2019; 123: 152-160.

CLSI. Performance standards for antimicrobial susceptibility testing: 15th informational supplement. CLSI document M100-S15PA, USA: Wayne. 2005.

Wold CW, Kjeldsen C, Corthay A, Rise F, Christensen BE, Duus JØ, Inngjerdingen KT. Structural characterization of bioactive heteropolysaccharides from the medicinal fungus Inonotus obliquus (Chaga). Carbohyd Polym. 2018; 185: 27-40.

Glamočlija J, Ćirić A, Nikolić M, Fernandes Â, Barros L, Calhelha RC, Ferreira I, Soković M, Van Griensven, L. (2015). Chemical characterization and biological activity of Chaga (Inonotus obliquus), a medicinal “mushroom”. J Ethnopharmacol. 2015; 162: 323-332.

Nakajima Y, Sato Y, Konishi T. Antioxidant Small Phenolic Ingredients in Inonotus obliquus (persoon) Pilat (Chaga). Chem. Pharm. Bull. 2007; 55(8): 1222-1226.

Lee I, Yun B. Styrylpyrone-class compounds from medicinal fungi Phellinus and Inonotus spp., and their medicinal importance. J Antibiot. 2011; 64(5): 349-359.

Zheng W, Miao K, Liu Y, Zhao Y, Zhang M, Pan S, Dai Y. Chemical diversity of biologically active metabolites in the sclerotia of Inonotus obliquus and submerged culture strategies for up-regulating their production. Appl. Microbiol. Biotechnol. 2010; 87(4): 1237-1254.

Burmasova MA, Utebaeva AA, Sysoeva EV, Sysoeva MA. Melanins of Inonotus obliquus: Bifidogenic and Antioxidant Properties. Biomolecules. 2019; 9(6): 248.

Kukulyanskaya TA, Kurchenko NV, Kurchenko VP, Babitskaya VG. Physicochemical Properties of Melanins Produced by the Sterile Form of Inonotus obliquus (“Chagi”) in Natural and Cultivated Fungus. Appl Biochem Micro+. 2002; 38(1): 58-61

Sysoeva MA, Yumaeva LR, Kuznetsova OY, Ziyatdinova GK, Budnikov GK, Melnikova, NB. Study of the composition of biologically active compounds in chaga meal. Perspectives of application of chaga meal in pharmaceutical industry. Russ. J. Gen. Chem. 2012; 82(3): 586-594.

Niu H, Song D, Mu H, Zhang W, Sun F, Duan J. Investigation of three lignin complexes with antioxidant and immunological capacities from Inonotus obliquus. Int. J. Biol. Macromol. 2016; 86: 587-593.

Ichimura T, Watanabe O, Maruyama S. Inhibition of HIV-1 Protease by Water-Soluble Lignin-Like Substance from an Edible Mushroom, Fuscoporia obliqua. Biosci. Biotechnol. Biochem. 1998; 62(3): 575-577.

Géry A, Dubreule C, André V, Rioult J, Bouchart V, Heutte N, De Pécoulas P, Krivomaz T, Garon D. Chaga (Inonotus obliquus), a Future Potential Medicinal Fungus in Oncology? A Chemical Study and a Comparison of the Cytotoxicity Against Human Lung Adenocarcinoma Cells (A549) and Human Bronchial Epithelial Cells (BEAS-2B). Integr. Cancer Ther. 2018; 17(3): 832-843.

Islam T, Yu X, Xu B. Phenolic profiles, antioxidant capacities and metal chelating ability of edible mushrooms commonly consumed in China. LWT. 2016; 72: 423-431.

Phaniendra A, Jestadi DB, Periyasamy L. Free Radicals: Properties, Sources, Targets, and Their Implication in Various Diseases. Indian J Clin Biochem. 2014; 30(1): 11-26.

Bray F, Ferlay J, Soerjomataram I, Siegel RL, Torre LA, Jemal A. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin. 2018; 68(6): 394-424.

Wang Z. Dietary polyphenols and colorectal cancer risk: The Fukuoka colorectal cancer study. World J Gastroenterol. 2013: 19(17): 2683-2690

Bahrami A, Jafari S, Rafiei P, Beigrezaei S, Sadeghi A, Hekmatdoost A, Rashidkhani B, Hejazi E. Dietary intake of polyphenols and risk of colorectal cancer and adenoma–A case-control study from Iran. Complement Ther Med. 2019; 45: 269-274.

Alam MN, Almoyad M, Huq F. Polyphenols in Colorectal Cancer: Current State of Knowledge including Clinical Trials and Molecular Mechanism of Action. Biomed Res Int. 2018; 2018: 1-29.

Blagodatski A, Yatsunskaya M, Mikhailova V, Tiasto V, Kagansky A, Katanaev VL. Medicinal mushrooms as an attractive new source of natural compounds for future cancer therapy. Oncotarget. 2018; 9(49): 29259-29274

Stojanović R, Belščak-Cvitanović A, Manojlović V, Komes D, Nedović V, Bugarski B. Encapsulation of thyme (Thymus serpyllum L.) aqueous extract in calcium-alginate beads. J Sci Food Agr. 2012; 92: 685–696.

González E, Gómez-Caravaca AM, Giménez B, Cebrián R, Maqueda M, Martínez-Férez A, Segura-Carretero A, Robert P. Evolution of the phenolic compounds profile of olive leaf extract encapsulated by spray-drying during in vitro gastrointestinal digestion. Food Chem. 2019; 279: 40–48.

Istenič K, Balanč B, Djordjević V, Bele M, Nedović V, Bugarski B, Poklar-Urlih N. Encapsulation of resveratrol into calcium-alginate submicron particles. J. Food Eng. 2015; 167: 196–203.

Ćujić N, Trifković K, Bugarski B, Ibrić S, Pljevljakušić D, Šavikin K. Chokeberry (Aronia melanocarpa L.) extract loaded in alginate and alginate/inulin system. Ind Crop Prod. 2016; 86: 120–131.