FARKLI SOLVENT SİSTEMLERİNİN KESTANE KABUKLARINDAN FENOLİK BİLEŞİKLERİN EKSTRAKSİYONU VE ANTİOKSİDAN AKTİVİTE ÜZERİNE ETKİSİ

Öz

Kestane kabukları genellikle atık olarak kabul edilmesine rağmen antioksidan kapasitesi yüksek fenolik bileşikler açısından zengindir. Bu çalışmada çeşitli çözücü sistemlerinin kestane kabuklarının biyoaktif içeriği [toplam fenolik (TPC), toplam flavonoid, toplam hidrolize tanen içerikleri ve antioksidan aktivite (DPPH, EC50)] üzerine etkileri incelenmiştir. Ekstraksiyonda etanol (E), saf aseton (A), su (W), etanol–aseton karışımı (E-A), %50 aseton (%50 A-W) ve %50 etanol (%50 E-W) kullanılmıştır. Ayrıca kestane kabuğunun organik ve amino asit profili belirlenmiştir. Elde edilen verilere göre; en yüksek toplam fenolik madde, flavonoid ve hidrolize tanen içerikleri sırasıyla %50 A-W ve %50 E-W ekstraktlarında belirlenmiş; saf aseton ekstraktı ise tüm parametreler bakımından en düşük değerleri vermiştir. Antioksidan aktiviteyi temsil eden EC50 değerleri açısından, en düşük değer %50 A-W ekstraktına ait olup bunu %50 E-W ve W ekstraktları takip etmiştir. Saf aseton ekstraktı, yüksek EC50 değeriyle en zayıf antioksidan kapasiteyi sergilemiştir. Kestane kabuğunda tespit edilen en baskın organik asit propiyonik asittir (3370.45 mg/kg). Amino asit profili açısından glutamik asit (0.399 g/100 g) ve aspartik asit (0.381 g/100 g) başlıca esansiyel olmayan amino asitler iken histidin (0.319 g/100 g) ve lisin (0.178 g/100 g) baskın esansiyel amino asitler olarak bulunmuştur. Elde edilen bulgular, kestane kabuğu ekstraktlarının yüksek bir in vitro antioksidan kapasiteye sahip olduğunu ve gerçek gıda sistemlerinde doğrulanmak kaydıyla, sentetik antioksidanlara doğal bir alternatif olabileceğini göstermektedir.

Anahtar Kelimeler

• Kestane kabuğu
• fenolik bileşikler
• flavonoid
• tanen
• solvent seçimi

IMPACT OF SOLVENT SYSTEM CHOICE ON THE RECOVERY OF PHENOLIC COMPOUNDS AND ANTIOXIDANT PROPERTIES IN EXTRACTS OF CHESTNUT SHELL

Öz

Chestnut shells, generally considered agricultural by-products, are rich in phenolic compounds with antioxidant properties. The current study investigates the influence of various solvent systems on bioactive contents [total phenolic content (TPC), total flavonoid content (TFC), total hydrolysable tannin content (TT), and antioxidant capacity (DPPH, EC50)]. Ethanol (E), acetone (A), water (W), ethanol-acetone (E-A), 50% acetone-water (50% A-W) and 50% ethanol-water (50% E-W) were used as extraction solvents. In addition, the organic and amino acid profiles of chestnut shells were determined. 50% A-W and 50% E-W extracts yielded the highest TPC, TFC and TT values, while pure acetone exhibited the lowest values for all three parameters. The lowest EC50 value was found in 50% A-W extract, followed by 50% E-W and W extracts. Among the quantified organic acids, propionic acid (3370.45 mg/kg) was the major compound in the chestnut shell. Regarding the amino acid profile, glutamic acid (0.399 g/100 g) and aspartic acid (0.381 g/100 g) were the major non-essential amino acids whereas histidine (0.319 g/100 g) and lysine (0.178 g/100 g) were predominant essential amino acids. The findings suggest that extracts from chestnut shells have a high in vitro antioxidant capacity and, subject to validation in real food systems, may serve as a natural alternative to synthetic antioxidants.

Anahtar Kelimeler

• Chestnut shell
• phenolic compounds
• flavonoids
• hydrolysable tannins
• solvent selection
• antioxidant activity

Kaynakça

1. Borges, O., Gonçalves, B., de Carvalho, J. L. S., Correia, P., Silva, A. P. (2008). Nutritional quality of chestnut (Castanea sativa Mill.) cultivars from Portugal. Food Chemistry, 106(3), 976-984. https://doi.org/10.1016/j.foodchem.2007.07.011
2. Cacciola, N. A., Squillaci, G., D’Apolito, M., Petillo O., Veraldi, F., La Gara, F. & Morana A. (2019). Castanea sativa Mill. shells aqueous extract exhibits anticancer properties inducing cytotoxic and pro-apoptotic effects. Molecules, 24(18), 3401. https://doi.org/10.3390/molecules24183401
3. Chambers, E., Viardot, A., Psichas, A., Douglas, M., Murphy, K. G., Zac-Varghese, S. E. K. & Frost, G. (2015). Effects of targeted delivery of propionate to the human colon on appetite regulation, body weight maintenance and adiposity in overweight adults. Gut, 64(11), 1744-1754. https://doi.org/10.1136/gutjnl-2014-307913
4. Chemat, F., Vian, M. A., Cravotto, G. (2012). Green extraction of natural products: Concept and principles. International Journal of Molecular Sciences, 13(7), 8615-8627. https://doi.org/10.3390/ijms13078615
5. Cravotto, C., Grillo, G., Binello, A., Gallina, L., Olivares-Vicente, Mariló., Herranz-López, M. & Cravotto, Giancarlo. Bioactive Antioxidant Compounds from Chestnut Peels through Semi-Industrial Subcritical Water Extraction. Antioxidants, 11(5), 988. https://doi.org/10.3390/antiox11050988
6. Dai, J., Mumper, R. J. (2010). Plant phenolics: extraction, analysis and their antioxidant and anticancer properties. Molecules, 15(10), 7313-7352. https://doi.org/10.3390/molecules15107313
7. Food and Agriculture Organization (FAO). (2024). https://www.fao.org/faostat/en/#data/QCL. Fernandez-Agullo, A., Freire, M. S., Antorrena, G., Pereira, J. A., Gonzalez-Alvarez, J. (2014). Effect of the extraction technique and operational conditions on the recovery of bioactive compounds from chestnut (Castanea sativa) bur and shell. Separation Science and Technology, 49(2), 267-277. https://doi.org/10.1080/01496395.2013.838264
8. Gul, O., Mortas, M., Atalar, I., Dervisoglu, M., Kahyaoglu, T. (2015). Manufacture and characterization of kefir made from cow and buffalo milk, using kefir grain and starter culture. Journal of Dairy Science, 98(3), 1517-1525. https://doi.org/10.3168/jds.2014-8755

9. Heim, K. E., Tagliaferro, A. R., Bobilya, D. J. (2002). Flavonoid antioxidants: chemistry, metabolism and structure-activity relationships. The Journal of Nutritional Biochemistry, 13(10), 572-584. https://doi.org/10.1016/S0955-2863(02)00208-5
10. Holeček, M. (2020). Histidine in health and disease: metabolism, physiological importance, and use as a supplement. Nutrients, 12(3), 848. https://doi.org/10.3390/nu12030848
11. Hu, M., Yang, X., Chang, X. (2021). Bioactive phenolic components and potential health effects of chestnut shell: A review. Journal of Food Biochemistry, 45(4), e13696.   https://doi.org/10.1111/jfbc.13696
12. Huang, M., Liao, C., Xie, J., Chen, J., Cao, F. (2023). Lysine content and its relationship with protein content in indica rice landraces of China. Food Chemistry: X, 17, 100549. https://doi.org/10.1016/j.fochx.2022.100549
13. Ignat, I., Volf, I., Popa, V. I. (2011). A critical review of methods for characterisation of polyphenolic compounds in fruits and vegetables. Food Chemistry, 126(4), 1821-1835. https://doi.org/10.1016/j.foodchem.2010.12.026
14. Karaman, S., Öztürk, G. (2025). Ultrasound-assisted extraction of phenolics from Spirulina platensis by different solvents: An optimization study based on simplex lattice mixture design. International Journal of Secondary Metabolite, 12(4), 772-780. https://doi.org/10.21448/ijsm.1554137
15. Lameirao, F., Pinto, D., Vieira, E. F., Peixoto, A. F., Freire, C., Sut, S. & Rodrigues, F. (2020). Green-sustainable recovery of phenolic and antioxidant compounds from industrial chestnut shells using ultrasound-assisted extraction: Optimization and evaluation of biological activities in vitro. Antioxidants, 9(3), 267. https://doi.org/10.3390/antiox9030267
16. Markom, M., Hasan, M., Daud, W. R. W., Singh, H., Jahim, J. M. (2007). Extraction of hydrolysable tannins from Phyllanthus niruri Linn.: Effects of solvents and extraction methods. Separation and Purification Technology, 52(3), 487-496. https://doi.org/10.1016/j.seppur.2006.06.003
17. Mustafa, A. M., Abouelenein, D., Acquaticci, L., Alessandroni, L., Abd-Allah, R. H., Borsetta, G. &, Caprioli, G. (2021). Effect of Roasting, Boiling, and Frying Processing on 29 Polyphenolics and Antioxidant Activity in Seeds and Shells of Sweet Chestnut (Castanea sativa Mill.). Plants, 10(10), 2192. https://doi.org/10.3390/plants10102192 
18. Panzella, L., Moccia, F., Nasti, R., Marzorati, S., Verotta, L., Napolitano, A. (2020). Bioactive phenolic compounds from agri-food wastes: An update on green and sustainable extraction methodologies. Frontiers in Nutrition, 7, 60. https://doi.org/10.3389/fnut.2020.00060
19. Pazara, M., Provelengiadi, G., Mantiniotou, M., Athanasiadis, V., Samanidis, I., Makrygiannis, I. &, Lalas, S. I. (2025). Optimizing Bioactive Compound Recovery from Chestnut Shells Using Pressurized Liquid Extraction and the Box–Behnken Design. Processes, 13(7), 2283. https://doi.org/10.3390/pr13072283 
20. Pérez-Jiménez, J., Arranz, S., Tabernero, M., Díaz-Rubio, M. E., Serrano, J., Goñi, I., Saura-Calixto, F. (2008). Updated methodology to determine antioxidant capacity in plant foods, oils and beverages: Extraction, measurement and expression of results. Food Research International, 41(3), 274-285. https://doi.org/10.1016/j.foodres.2007.12.004
21. Pinto, D., de la Luz Cadiz-Gurrea, M., Garcia, J., Saavedra, M. J., Freitas, V., Costa, P. &, Rodrigues, F. (2021a). From soil to cosmetic industry: Validation of a new cosmetic ingredient extracted from chestnut shells. Sustainable Materials and Technologies, 29, e00309. https://doi.org/10.1016/j.susmat.2021.e00309
22. Pinto, D., Vieira, E. F., Peixoto, A. F., Freire, C., Freitas, V., Costa, P., Rodrigues, F. (2021b). Optimizing the extraction of phenolic antioxidants from chestnut shells by subcritical water extraction using response surface methodology. Food Chemistry, 334, 127521. https://doi.org/10.1016/j.foodchem.2020.127521
23. Rice-Evans, C. A., Miller, N. J., Paganga, G. (2005). Structure–Antioxidant Activity Relationships of Flavonoids and Phenolic Acids. Free Radical Biology and Medicine, 39(10), 1266. https://doi.org/10.1016/0891-5849(95)02227-9
24. Rodrigues, D. B., Verissimo, L., Finimundy, T., Rodrigues, J., Oliveira, I., Gonçalves, J. & Calhelha, R. C. (2023). Chemical and bioactive screening of green polyphenol-rich extracts from chestnut by-products: an approach to guide the sustainable production of high-added value ingredients. Foods, 12(13), 2596. https://doi.org/10.3390/foods12132596
25. Rodrigues, F., Santos, J., Pimentel, F. B., Braga, N., Palmeira-de-Oliveira, A., Oliveira, M. B. P. (2015). Promising new applications of Castanea sativa shell: Nutritional composition, antioxidant activity, amino acids and vitamin E profile. Food & Function, 6(8), 2854-2860. https://doi.org/10.1039/C5FO00571J
26. Shahidi, F., Ambigaipalan, P. (2015). Phenolics and polyphenolics in foods, beverages and spices: Antioxidant activity and health effects–A review. Journal of Functional Foods, 18, 820-897. https://doi.org/10.1016/j.jff.2015.06.018
27. Silva, V., Falco, V., Dias, M. I., Barros, L., Silva, A., Capita, R. & Ferreira, I. C. F. R. (2020). Evaluation of the phenolic profile of Castanea sativa Mill. by-products and their antioxidant and antimicrobial activity against multiresistant bacteria. Antioxidants, 9(1), 87. https://doi.org/10.3390/antiox9010087
28. Singleton, V. L., Rossi, J. A. (1965). Colorimetry of total phenolics with phosphomolybdic-phosphotungstic acid reagents. American Journal of Enology and Viticulture, 16(3), 144-158. https://doi.org/10.5344/ajev.1965.16.3.144 Smeriglio, A., Barreca, D., Bellocco, E., Trombetta, D. (2017). Proanthocyanidins and hydrolysable tannins: occurrence, dietary intake and pharmacological effects. British Journal of Pharmacology, 174(11), 1244-1262. https://doi.org/10.1111/bph.13630
29. Sorice, A., Siano, F., Capone, F., Guerriero, E., Picariello, G., & ,Volpe, M. G. (2016). Potential anticancer effects of polyphenols from chestnut shell extracts: Modulation of cell growth, and cytokinomic and metabolomic profiles. Molecules, 21(10), 1411. https://doi.org/10.3390/molecules21101411
30. Speisky, H., Shahidi, F., Costa de Camargo, A., Fuentes, J. (2022). Revisiting the oxidation of flavonoids: Loss, conservation or enhancement of their antioxidant properties. Antioxidants, 11(1), 133. https://doi.org/10.3390/antiox11010133
31. Squillaci, G., Apone, F., Sena, L. M., Carola, A., Tito, A., Bimonte, M. & Morana, A. (2018). Chestnut (Castanea sativa Mill.) industrial wastes as a valued bioresource for the production of active ingredients. Process Biochemistry, 64, 228-236. https://doi.org/10.1016/j.procbio.2017.09.017
32. Sun, X., Feng, R., Li, Y., Lin, S., Zhang, W., &, Li, S. (2014). Histidine supplementation alleviates inflammation in the adipose tissue of high-fat diet-induced obese rats via the NF-κB-and PPARγ-involved pathways. British Journal of Nutrition, 112(4), 477-485. https://doi.org/10.1017/S0007114514001056
33. Vallejo, M. J., Salazar, L., Grijalva, M. (2017). Oxidative stress modulation and ROS‐mediated toxicity in cancer: a review on in vitro models for plant‐derived compounds. Oxidative Medicine and Cellular Longevity, 2017(1), 4586068.https://doi.org/10.1155/2017/4586068
34. Willis, R. B., Allen, P. R. (1998). Improved method for measuring hydrolyzable tannins using potassium iodate. Analyst, 123(3), 435-439. https://doi.org/10.1039/A706862J
35. Zhishen, J., Mengcheng, T., Jianming, W. (1999). The determination of flavonoid contents in mulberry and their scavenging effects on superoxide radicals. Food Chemistry, 64(4), 555-559. https://doi.org/10.1016/S0308-8146(98)00102-2