Vaccinium myrtillus’un İki Morfolojik Varyantının Karşılaştırmalı FTIR Profillemesi ve Antioksidan Potansiyeli

Abstract

Vaccinium myrtillus L. (çobanüzümü), zengin fitokimyasal bileşimi ve dikkat çekici biyolojik aktiviteleriyle bilinen yabani bir bitkidir. Bu çalışmada, V. myrtillus'un iki morfolojik varyantının farklı bitki kısımlarından (yaprak, meyve ve gövde) elde edilen metanol ekstrelerinin antioksidan kapasiteleri ve kimyasal profilleri açısından karşılaştırmalı olarak değerlendirilmiştir. Antioksidan aktiviteler, CUPRAC (Bakır İyonu İndirgeyici Antioksidan Kapasitesi) ve FRAP (Demir İndirgeyici Antioksidan Gücü) testleri kullanılarak belirlenirken, fonksiyonel grup karakterizasyonu FTIR-ATR spektroskopisi ile gerçekleştirilmiştir. Sonuçlar, antioksidan kapasitelerinin bitki kısmına ve varyantına bağlı olarak önemli ölçüde değiştiğini ortaya koymuştur. En yüksek antioksidan aktivite Yer Ligarbası’nın dal kısmında (CUPRAC'ta 268.40 mg troloks eşdeğeri (TE)/g; FRAP'ta 139.29 µmol Fe²⁺/g) gözlemlenirken, en düşük değerler yine yer ligarbasının meyvesinde kaydedilmiştir. CUPRAC ve FRAP testleri arasındaki tutarlı eğilim, iki elektron transferine dayalı yöntem arasında güçlü bir uyum olduğunu göstermiştir. FTIR spektrumları, O–H (~3400 cm^-1), C–H (~2925 cm^-1), aromatik C=C (~1600 cm^-1) ve C–O (~1200–1000 cm^-1) titreşimleri de dahil olmak üzere fenolik bileşiklere, flavonoitlere, karbonhidratlara ve organik asitlere karşılık gelen karakteristik bantlar göstermiştir. Antioksidan kapasite ile 3400, 1020 ve 1050 cm^-1'deki FTIR bantları arasında orta düzeyde pozitif korelasyonlar belirlenmiştir; bu da hidroksil ve C–O içeren fenolik bileşiklerin antioksidan aktivitede önemli bir rol oynadığını düşündürmektedir. Genel olarak, bulgular V. myrtillus'un antioksidan potansiyelinin fenolik bileşimiyle güçlü bir şekilde ilişkili olduğunu ve bitki kısmına ve morfolojik varyantına bağlı olarak değiştiğini göstermektedir. FTIR analizleri, antioksidan aktivite ile ilişkili fenolik ve hidroksil içeren fonksiyonel grupların varlığını ortaya koymuş ve elde edilen CUPRAC ile FRAP sonuçlarını yapısal açıdan desteklemiştir.

Keywords

• Vaccinium myrtillus L.
• CUPRAC
• FRAP
• FTIR-ATR spektroskopisi

Supporting Institution

Bu çalışma için herhangi bir finansman alınmamıştır.

Ethical Statement

Etik beyan gerekmemektedir.

Thanks

Yazar, bu çalışmada kullanılan bitki ekstrelerini sağladıkları için Yuca ve arkadaşlarına minnettar olduklarını belirtirler; bu ekstreler daha önce yayınlanmış araştırmalarında hazırlanmıştır.

Comparative FTIR Profiling and Antioxidant Potential of Two Morphological Variants of Vaccinium myrtillus

Abstract

Vaccinium myrtillus L. is a wild plant known for its rich phytochemical composition and remarkable biological activities. In this study, methanol extracts obtained from different plant parts (leaves, fruits, and stems) of two morphological variants of V. myrtillus were comparatively evaluated in terms of their antioxidant capacities and chemical profiles. Antioxidant activities were determined using the CUPRAC (Cupric Ion Reducing Antioxidant Capacity) and FRAP (Ferric Reducing Antioxidant Power) assays, while functional group characterization was performed by FTIR-ATR spectroscopy. The results revealed that antioxidant capacities varied significantly depending on both the plant part and the morphological variant. The highest antioxidant activity was observed in the stem part of Yer Ligarbası (268.40 mg Trolox equivalent (TE)/g in the CUPRAC assay and 139.29 µmol Fe²⁺/g in the FRAP assay), whereas the lowest values were recorded in the fruit part of the same variant. The consistent trends obtained from the CUPRAC and FRAP assays indicated a strong agreement between these two electron transfer-based methods. FTIR spectra exhibited characteristic bands corresponding to phenolic compounds, flavonoids, carbohydrates, and organic acids, including O–H (~3400 cm^-1), C–H (~2925 cm^-1), aromatic C=C (~1600 cm^-1), and C–O (~1200–1000 cm^-1) vibrations. Moderate positive correlations were identified between antioxidant capacity and the FTIR bands at 3400, 1020, and 1050 cm^-1, suggesting that phenolic compounds containing hydroxyl and C–O functional groups play a significant role in antioxidant activity. Overall, the findings demonstrated that the antioxidant potential of V. myrtillus is strongly associated with its phenolic composition and varies according to plant part and morphological variant. Furthermore, FTIR spectroscopy was confirmed to be a rapid and effective complementary tool for evaluating the chemical profile and antioxidant potential of plant extracts. 

Keywords

• Vaccinium myrtillus L.
• CUPRAC
• FRAP
• FTIR-ATR spectroscopy

Supporting Institution

No funding was received for this study.

Ethical Statement

No ethical declaration is required.

Thanks

The author express gratitude to Yuca et al. for providing the plant extracts used in this study; these extracts were prepared in their previously published research.

References

1. Apak, R., Güçlü, K., Özyürek, M., & Karademir, S. E. (2004). Novel total antioxidant capacity index for dietary polyphenols and vitamins C and E, using their cupric ion reducing capability in the presence of neocuproine: CUPRAC method. Journal of agricultural and food chemistry, 52(26), 7970-7981. https://doi.org/10.1021/jf048741x
2. Ashour, O. M., Elberry, A. A., Alahdal, A. M., Al Mohamadi, A. M., Nagy, A. A., Abdel-Naim, A. B., Mohamadin, A. M. (2011). Protective effect of bilberry (Vaccinium myrtillus) against doxorubicin-induced oxidative cardiotoxicity in rats. Medical science monitor: international medical journal of experimental and clinical research, 17(4), BR110. https://doi.org/10.12659/MSM.881711
3. Azmir, J., Zaidul, I. S. M., Rahman, M. M., Sharif, K. M., Mohamed, A., Sahena, F., ... & Omar, A. K. (2013). Techniques for extraction of bioactive compounds from plant materials: A review. Journal of food engineering, 117(4), 426-436.
4. Bardakçı, B., & Seçilmiş, H. (2006). Isparta bölgesindeki gül yağinin kimyasal içeriğinin GC-MS ve FTIR spektroskopisi tekniği ile incelenmesi. Süleyman Demirel University Faculty of Arts and Science Journal of Science, 1(1), 64-69. https://doi.org/10.1016/j.jfoodeng.2013.01.014
5. Barnes, M., Sulé-Suso, J., Millett, J., & Roach, P. (2023). Fourier transform infrared spectroscopy as a non-destructive method for analysing herbarium specimens. Biology Letters, 19(3). https://doi.org/10.1098/rsbl.2022.0546
6. Batinić, P., Jovanović, A. A., Rajab Elferjane, M., Čutović, N., Milošević, M., Marinković, A., & Bugarski, B. (2023). Comparison of maceration and ultrasound-assisted extraction of antioxidant compounds from Vaccinium myrtillus L. Paper presented at the Proceedings-VIII International Congress" Engineering, Environment and Materials in Process Industry", EEM 2023, March 20-23 2023, Jahorina.
7. Benzie, I. F., & Strain, J. J. (1996). The ferric reducing ability of plasma (FRAP) as a measure of “antioxidant power”: the FRAP assay. Analytical biochemistry, 239(1), 70-76. https://doi.org/10.1006/abio.1996.0292
8. Ceriello, A., & Genovese, S. (2016). Atherogenicity of postprandial hyperglycemia and lipotoxicity. Reviews in Endocrine and Metabolic Disorders, 17(1), 111-116. https://doi.org/10.1007/s11154-016-9341-8

9. Chen, S., Peng, H. M., & Webster, R. D. (2010). Infrared and UV–vis spectra of phenoxonium cations produced during the oxidation of phenols with structures similar to vitamin E. Electrochimica acta, 55(28), 8863-8869. https://doi.org/10.1016/j.electacta.2010.07.096
10. Civaş, A., Karakaş, Ş., Bayrak, B., Akyüz, B., Keçeci, E. B., Özer, E. B., . . . Yuca, H. (2026). Comprehensive characterization of 26 Turkish asteraceae species: Insights from FTIR, phytochemical composition, and elemental analysis via ICP-MS. South African Journal of Botany, 189, 387-402. https://doi.org/10.1016/j.sajb.2025.12.010
11. Colak, N., Torun, H., Gruz, J., Strnad, M., Hermosín-Gutiérrez, I., Hayirlioglu-Ayaz, S., & Ayaz, F. A. (2016). Bog bilberry phenolics, antioxidant capacity and nutrient profile. Food chemistry, 201, 339-349. https://doi.org/10.1016/j.foodchem.2016.01.062
12. 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
13. David, I., Stefanut, M., Cata, A., Ienascu, I., Pop, R., Tanasie, C., & Balcu, I. (2009). Study of anthocyanins from Vaccinium myrtillus L. frozen fruits. Journal of Agroalimentary Processes and Technologies, 15(3), 348-352.
14. Dróżdż, P., Šėžienė, V., & Pyrzynska, K. (2017). Phytochemical properties and antioxidant activities of extracts from wild blueberries and lingonberries. Plant Foods for Human Nutrition, 72(4), 360-364. https://doi.org/10.1007/s11130-017-0640-3
15. Erlund, I., Koli, R., Alfthan, G., Marniemi, J., Puukka, P., Mustonen, P., . . . Jula, A. (2008). Favorable effects of berry consumption on platelet function, blood pressure, and HDL cholesterol. The American journal of clinical nutrition, 87(2), 323-331. https://doi.org/10.1093/ajcn/87.2.323
16. Fuente, E., Menéndez, J., Díez, M., Suárez, D., & Montes-Morán, M. (2003). Infrared spectroscopy of carbon materials: a quantum chemical study of model compounds. The Journal of physical chemistry B, 107(26), 6350-6359. https://doi.org/10.1021/jp027482g
17. Granfeldt, Y. E., & Björck, I. M. (2011). A bilberry drink with fermented oatmeal decreases postprandial insulin demand in young healthy adults. Nutrition Journal, 10(1), 57. https://doi.org/10.1186/1475-2891-10-57
18. Irawan, C., Dwi Putri, I., & Sukiman, M. (2022). Antioxidant activity of DPPH, CUPRAC, and FRAP methods, as well as activity of alpha-glucosidase inhibiting enzymes from Tinospora crispa (L.) stem ultrasonic extract.
19. Karcheva-Bahchevanska, D., Nikolova, M., & Iliev, I. (2023). Inhibitory Potential of Different Bilberry (Vaccinium myrtillus L.) Extracts on Human Salivary α-Amylase. Molecules, 28(15), 5820. https://doi.org/10.3390/molecules28155820
20. Kolehmainen, M., Mykkänen, O., Kirjavainen, P. V., Leppänen, T., Moilanen, E., Adriaens, M., . . . Pulkkinen, L. (2012). Bilberries reduce low‐grade inflammation in individuals with features of metabolic syndrome. Molecular nutrition & food research, 56(10), 1501-1510. https://doi.org/10.1002/mnfr.201200195
21. Kubilienė, A., Marksa, M., Baranauskaitė, J., Ragažinskienė, O., & Ivanauskas, L. (2020). Comparative evaluation of antioxidant activity of Cannabis sativa L. using FRAP and CUPRAP assays. Chemija, 31(3). https://doi.org/10.6001/chemija.v31i3.4289
22. Luczaj, L., Pieroni, A., Tardío, J., Pardo-de-Santayana, M., Sõukand, R., Svanberg, I., & Kalle, R. (2012). Wild food plant use in 21st century Europe: the disappearance of old traditions and the search for new cuisines involving wild edibles. Acta societatis botanicorum poloniae, 81(4), 359-370. https://dx.doi.org/10.5586/asbp.2012.031
23. Luo, H., Lv, X.-D., Wang, G.-E., Li, Y.-F., Kurihara, H., & He, R.-R. (2014). Anti-inflammatory effects of anthocyanins-rich extract from bilberry (Vaccinium myrtillus L.) on croton oil-induced ear edema and Propionibacterium acnes plus LPS-induced liver damage in mice. International journal of food sciences and nutrition, 65(5), 594-601. https://doi.org/10.3109/09637486.2014.886184
24. Luteyn, J. L. (2002). Diversity, adaptation, and endemism in neotropical Ericaceae: biogeographical patterns in the Vaccinieae. The Botanical Review, 68(1), 55-87. https://doi.org/10.1663/0006-8101(2002)068[0055:DAAEIN]2.0.CO;2
25. Mazza, G., & Miniati, E. (2018). Anthocyanins in fruits, vegetables, and grains: CRC press. https://doi.org/10.1201/9781351069700
26. Nguyen, V., Tang, J., Oroudjev, E., Lee, C. J., Marasigan, C., Wilson, L., & Ayoub, G. (2010). Cytotoxic effects of bilberry extract on MCF7-GFP-tubulin breast cancer cells. Journal of Medicinal food, 13(2), 278-285. https://doi.org/10.1089/jmf.2009.0053
27. Novi, S., Caponigro, V., Miranda, M. R., Aquino, G., Carri, M. D., Salviati, E., ... & Manfra, M. (2025). Metabolomics insights into the protective molecular mechanism of Vaccinium myrtillus against oxidative stress in intestinal cells. Scientific Reports, 15(1), 8643. https://doi.org/10.1038/s41598-025-93722-x
28. Pacifici, R. E., & Davies, K. J. (1991). Protein, lipid and DNA repair systems in oxidative stress: the free-radical theory of aging revisited. Gerontology, 37(1-3), 166-180. https://doi.org/10.1159/000213257
29. Patra, J. K., Shin, H. S., & Das, G. (2026). A Review of Major Compounds in Bilberry (Vaccinium myrtillus L.) Fruits and Leaves: Isolation, Purification, and Their Antiaging Effects. Nutrients, 18(2), 350. https://doi.org/10.3390/nu18020350
30. Pardo-de-Santayana, M., Tardío, J., Blanco, E., Carvalho, A. M., Lastra, J. J., San Miguel, E., & Morales, R. (2007). Traditional knowledge of wild edible plants used in the northwest of the Iberian Peninsula (Spain and Portugal): a comparative study. Journal of ethnobiology and ethnomedicine, 3(1), 27. https://doi.org/10.1186/1746-4269-3-27
31. Pieroni, A., Cianfaglione, K., Nedelcheva, A., Hajdari, A., Mustafa, B., & Quave, C. L. (2014). Resilience at the border: traditional botanical knowledge among Macedonians and Albanians living in Gollobordo, Eastern Albania. Journal of Ethnobiology and Ethnomedicine, 10(1), 31. https://doi.org/10.1186/1746-4269-10-31
32. Pires, T. C., Caleja, C., Santos-Buelga, C., Barros, L., & Ferreira, I. C. (2020). Vaccinium myrtillus L. fruits as a novel source of phenolic compounds with health benefits and industrial applications-a review. Current pharmaceutical design, 26(16), 1917-1928. https://doi.org/10.2174/1381612826666200317132507
33. Pizzino, G., Irrera, N., Cucinotta, M., Pallio, G., Mannino, F., Arcoraci, V., . . . Bitto, A. (2017). Oxidative stress: harms and benefits for human health. Oxidative medicine and cellular longevity, 2017(1), 8416763. https://doi.org/10.1155/2017/8416763
34. Poja, R., Ştefan, I.-D., Oprinescu, C.-I., Hădărugă, D.-I., & Hădărugă, N.-G. (2025). Autochthonous Vaccinium Myrtillus L.–Antioxidant Activity, Kinetics And Cyclodextrin Nanoencapsulation Of The Hydrophilic Extracts. International Multidisciplinary Scientific GeoConference: SGEM, 6(1), 3-10. https://doi.org/10.5593/sgem2025/6.1/s23.01
35. Routray, W., & Orsat, V. (2011). Blueberries and their anthocyanins: factors affecting biosynthesis and properties. Comprehensive Reviews in Food Science and Food Safety, 10(6), 303-320. https://doi.org/10.1111/j.1541-4337.2011.00164.x
36. Saral, Ö., Ölmez, Z., & Şahin, H. (2015). Comparison of antioxidant properties of wild blueberries (Vaccinium arctostaphylos L. and Vaccinium myrtillus L.) with cultivated blueberry varieties (Vaccinium corymbosum L.) in Artvin region of Turkey. Turkish Journal of Agriculture-Food Science and Technology, 3(1), 40-44.
37. Schink, A., Neumann, J., Leifke, A. L., Ziegler, K., Fröhlich-Nowoisky, J., Cremer, C., . . . Schuppan, D. (2018). Screening of herbal extracts for TLR2-and TLR4-dependent anti-inflammatory effects. PloS one, 13(10), e0203907. https://doi.org/10.1371/journal.pone.0203907
38. Sethi, S., Joshi, A., Arora, B., Bhowmik, A., Sharma, R., & Kumar, P. (2020). Significance of FRAP, DPPH, and CUPRAC assays for antioxidant activity determination in apple fruit extracts. European Food Research and Technology, 246(3), 591-598. https://doi.org/10.1007/s00217-020-03432-z
39. Sezer, E. D., Oktay, L. M., Karadadaş, E., Memmedov, H., Selvi Gunel, N., & Sözmen, E. (2019). Assessing anticancer potential of blueberry flavonoids, quercetin, kaempferol, and gentisic acid, through oxidative stress and apoptosis parameters on HCT-116 cells. Journal of medicinal food, 22(11), 1118-1126. https://doi.org/10.1089/jmf.2019.0098
40. Sies, H. (2015). Oxidative stress: a concept in redox biology and medicine. Redox biology, 4, 180-183. https://doi.org/10.1016/j.redox.2015.01.002
41. Sinelli, N., Spinardi, A., Di Egidio, V., Mignani, I., & Casiraghi, E. (2008). Evaluation of quality and nutraceutical content of blueberries (Vaccinium corymbosum L.) by near and mid-infrared spectroscopy. Postharvest Biology and Technology, 50(1), 31-36. https://doi.org/10.1016/j.postharvbio.2008.03.013
42. Thibado, S. P., Thornthwaite, J. T., Ballard, T. K., & Goodman, B. T. (2018). Anticancer effects of Bilberry anthocyanins compared with NutraNanoSphere encapsulated Bilberry anthocyanins. Molecular and clinical oncology, 8(2), 330-335. https://doi.org/10.3892/mco.2017.1520
43. Yuca, H., Ayar, B., Aydın, B., Çoban, F., & Karakaya, S. (2025). In vitro evaluation of antidiabetic, antioxidant, and anticholinergic properties of different parts of Bilberry (Vaccinium myrtillus L.) from the Eastern black sea Region, with LC/MS–MS analysis of their chemical composition in food applications. Food Science & Nutrition, 13(5), e70249. https://doi.org/10.1002/fsn3.70249
44. Žiberna, L., Lunder, M., Može, Š., Vanzo, A., & Drevenšek, G. (2009). Cardioprotective effects of bilberry extract on ischemia-reperfusion-induced injury in isolated rat heart. BMC Pharmacology, 9(Suppl 2), A55. https://doi.org/10.1186/1471-2210-9-S2-A55