Trifolium pratense L. gövdesinin kantitatif analizi ve antioksidan aktivitesi

Yıl 2025, Cilt: 6 Sayı: 1, 1 - 8, 30.04.2025

Ramazan Erenler , İlyas Yıldız , Esma Nur Geçer , İbrahim Hosaflıoğlu , Süleyman Muhammed Çelik

https://doi.org/10.53445/batd.1669871

Öz

Doğal ürünlerin tıbbi ve beslenme amaçlı kullanımı çok eski çağlara dayanmakta olup, içerdikleri bileşikler sayesinde ilaçların keşfi ve geliştirilmesine önemli katkılarda bulunmuştur. Bu çalışmada, Trifolium pratense gövdesinin fenolik bileşiklerinin kantitatif analizi ve antioksidan aktivitesi belirlenmiştir. Rutin (0,075 mg/g ekstre), salisilik asit (0,044), trans-ferulik asit (0,0385), gallik asit (0,0329) ve hesperidin (0,024) ana ürün olarak belirlenmiştir. Sapın metanol ekstresinin antioksidan aktivitesi DPPH ve ABTS analizleri kullanılarak değerlendirilmiştir. DPPH analizinde ekstre, 17,22 ± 0,67 µg/mL’lik bir IC50 değeriyle orta düzeyde aktivite göstermiştir. BHA ve BHT aktiviteleri sırasıyla (5.47 ± 0.23, IC50, µg/mL) ve (10.67 ± 0.20, IC50, µg/mL) olarak hesaplandı. ABTS aktivitesine gelince, ekstre yüksek aktivite gösterdi (IC50, 9.03 ± 0.11 µg/mL). Standart BHT’nin aktivitesi 9.51 ± 0.25 µg/mL (IC50) olarak tespit edildi. Trifolium pratense sapı, ilgili bitkilerden izole edilebilen değerli biyoaktif bileşik kaynağı ve antioksidan olabilir.

Anahtar Kelimeler

Trifolium pratense L. gövde , LC-MS/MS , antioksidan aktivite

Quantitative analysis and antioxidant activity of Trifolium pratense L. stem

Öz

The use of natural products for medical and nutritional purposes dates back to ancient times and has made significant contributions to the discovery and development of drugs, thanks to the compounds they contain. In this study, quantitative analysis of phenolic compounds and antioxidant activity of Trifolium pratense stem were determined. Rutin (0.075 mg/g extract), salicylic acid (0.044 mg/g extract), trans-ferulic acid (0.004 mg/g extract), gallic acid (0.003 mg/g extract), and hesperidin (0.024 mg/g extract) were determined as major compounds. The antioxidant activity of the methanol extract of the stem was evaluated using the DPPH and ABTS assays. In the DPPH assay, the extract exhibited moderate activity with an IC50 value of 17.22 ± 0.67 µg/mL. The activities of BHA and BHT were calculated as (5.47 ± 0.23, IC50, µg/mL) and (10.67 ± 0.20, IC50, µg/mL), respectively. Regarding the ABTS activity, the extract displayed high activity (IC50, 9.03 ± 0.11 µg/mL). The standard BHT activity was detected as 9.51 ± 0.25 µg/mL (IC50). Trifolium pratense stem could be a valuable source of antioxidant and bioactive compounds that can be isolated from the corresponding plants.

Anahtar Kelimeler

Trifolium pratense L. stem , LC-MS/MS , antioxidant activity

Kaynakça

• Aissous, I., Benrebai, M., Ameddah, S., Menad, A., Erenler, R., Benayache, S., & Benayache, F. (2023). The preventive effects of Centaurea maroccana Ball. extract against oxidative stress induced by cisplatin in mice brains: in vitro and in vivo studies. Drug and Chemical Toxicology, 46(6), 1162-1175. https://doi.org/10.1080/01480545.2022.2139841
• Aksit, H., Çelik, S. M., Sen, Ö., Erenler, R., Demirtas, I., Telci, I., & Elmastas, M. (2014). Complete isolation and characterization of polar portion of Mentha dumetorum water extract. Records of Natural Products, 8(3), 277-280.
• Atalar, M. N., Erenler, R., Turkan, F., Alma, M. H., Demirtas, I., Baran, A., Kandemir, S. I., Kekec, A. I., & Saltan, F. Z. (2023). Phytochemical analysis and biological activity of Corchorus olitorius L.: Quantitative analysis of bioactive compounds by LC–MS/MS, antibacterial, enzyme inhibition, and cytotoxic activities. European Journal of Integrative Medicine, 62, 102290. https://doi.org/10.1016/j.eujim.2023.102290
• Bakchiche, B., Hadjira, G., Erenler, R., Yildiz, I., Asmaa, S. M., & El-shazly, M. A. (2024). Mineral contents, Antioxidant and Antimicrobial Activities of Algerian Terfezia claveryi extracts. [Cezayir Terfezia claveryi ekstraktlarının mineral içerikleri, Antioksidan ve Antimikrobiyal Aktiviteleri]. Fabad Journal of Pharmaceutical Sciences, 49(1), 19-36. https://doi.org/10.55262/fabadeczacilik.1326710
• Başar, Y., & Erenler, R. (2024). Phytochemical analysis of Silybum marianum flowers: Quantitative analysis of natural compounds and molecular docking application. [Silybum marianum çiçeklerinin fitokimyasal analizi: Doğal bileşiklerin kantitatif analizi ve moleküler yerleştirme uygulaması]. Turkish Journal of Biodiversity, 7(1), 20-31. https://doi.org/10.38059/biodiversity.1450643
• Bayir, B., Gündüz, H., Usta, T., Şahin, E., Özdemir, Z., Kayır, Ö., Sen, Ö., Akşit, H., Elmastaş, M., & Erenler, R. (2014). Chemical Composition of Essential Oil from Marrubium Vulgare L. Leaves. Journal of New Results in Science, 6(6), 44-50.
• Cragg, G. M., Newman, D. J., & Snader, K. M. (1997). Natural products in drug discovery and development. Journal of Natural Products, 60(1), 52-60. https://doi.org/10.1021/np9604893
• Çelik, I., Erenler, R., Akkurt, M., Ünlü, C. S., Tutar, A., & Garcia-Granda, S. (2007). Crystal structure of 1,2,3-tribromobenz[f]indane. Analytical Sciences: X-ray Structure Analysis Online, 23(11), x229-x230. Retrieved from https://www.scopus.com/inward/record.uri?eid=2-s2.0-70549106673&partnerID=40&md5=6742016390819e9a0382e6e7a59ef428
• Dag, B. (2022). Green synthesis, characterization, and antioxidant activity of silver nanoparticles using Stachys annua L. subsp. annua var. annua. Particulate Science and Technology, 40(4), 512-520. https://doi.org/10.1080/02726351.2021.1966689
• Demirtas, I., Erenler, R., Elmastas, M., & Goktasoglu, A. (2013). Studies on the antioxidant potential of flavones of Allium vineale isolated from its water-soluble fraction. Food Chemistry, 136(1), 34-40. https://doi.org/10.1016/j.foodchem.2012.07.086
• Elmastas, M., Ozturk, L., Gokce, I., Erenler, R., & Aboul‐Enein, H. Y. (2004). Determination of antioxidant activity of marshmallow flower (Althaea officinalis L.). Analytical Letters, 37(9), 1859-1869. https://doi.org/10.1081/AL-120039431
• Erenler, R., Atalar, M. N., Yıldız, İ., Geçer, E. N., Yıldırım, A., Demirtas, İ., & Alma, M. H. (2023). Quantitative analysis of bioactive compounds by LC-MS/MS from Inula graveolens. Bütünleyici ve Anadolu Tıbbı Dergisi, 4(3), 3-10. https://doi.org/10.53445/batd.1278048
• Erenler, R., & Biellmann, J. F. (2007). Synthesis of pentafluorophenyl- and pyridinyl-3 allenes. Journal of the Chinese Chemical Society, 54(1), 103-108. https://doi.org/10.1002/jccs.200700017
• Erenler, R., & Dag, B. (2022). Biosynthesis of silver nanoparticles using Origanum majorana L. and evaluation of their antioxidant activity. Inorganic and Nano-Metal Chemistry, 52(4), 485-492. https://doi.org/10.1080/24701556.2021.1952263
• Erenler, R., Demirtas, I., Buyukkidan, B., & Cakmak, O. (2006). Synthesis of hydroxy, epoxy, nitrato and methoxy derivatives of tetralins and naphthalenes. Journal of Chemical Research(12), 753-757. https://doi.org/10.3184/030823406780199721
• Erenler, R., Demirtas, I., Karan, T., Gul, F., Kayir, O., & Karakoc, O. C. (2018). Chemical constituents, quantitative analysis and insecticidal activities of plant extract and essential oil from Origanum onites L. Trends in Phytochemical Research, 2(2), 91-96.
• Erenler, R., & Gecer, E. N. (2022a). Green synthesis of silver nanoparticles from Astragalus logopodioides L. leaves. Turkish Journal of Agriculture-Food Science and Technology, 10(6), 1112-1115. https://doi.org/10.24925/turjaf.v10i6.1112-1115.5190
• Erenler, R., & Gecer, E. N. (2022b). Synthesis of Silver Nanoparticles Using Sideritis montana L. Leaf Extract: Characterization, Catalytic Degradation of Methylene Blue and Antioxidant Activity. Journal of Nano Research, 75, 17-28. https://doi.org/10.4028/p-333bjm
• Erenler, R., Gecer, E. N., Genc, N., & Yanar, D. (2021). Antioxidant activity of silver nanoparticles synthesized from Tagetes erecta L. leaves. International Journal of Chemistry and Technology, 5(2), 141-146. https://doi.org/10.32571/ijct.1005275
• Erenler, R., Geçer, E. N., Alma, M. H., & Demirtas, İ. (2022). Quantitative Analysis of Bioactive Compounds by High‑Performance Liquid Chromatography in Origanum bilgeri. Journal of Integrative and Anatolian Medicine, 4(1), 15-20. https://doi.org/10.53445/batd.1210373
• Erenler, R., & Hosaflioglu, I. (2023). Green synthesis of silver nanoparticles using Onobrychis sativa L.: Characterization, catalytic degradation of methylene blue, antioxidant activity, and quantitative analysis of bioactive compounds. Materials Today Communications, 35, 105863. https://doi.org/10.1016/j.mtcomm.2023.105863
• Erenler, R., Karan, T., & Hosaflioglu, İ. (2023). Phytochemical analysis of Syringa vulgaris: Quantitative analysis of natural compounds by LC-ESI-MS/MS. Turkish Journal of Biodiversity, 6(2), 75-78. https://doi.org/10.38059/biodiversity.1312872
• Erenler, R., Telci, İ., Elmastaş, M., Akşit, H., Gül, F., Tüfekçi, A. R., Demirtaş, İ., & Kayir, Ö. (2018). Quantification of flavonoids isolated from Mentha spicata in selected clones of Turkish mint landraces. Turkish Journal of Chemistry, 42(6), 1695-1705. https://doi.org/10.3906/kim-1712-3
• Erenler, R., Telci, I., Ulutas, M., Demirtas, I., Gul, F., Elmastas, M., & Kayir, O. (2015). Chemical Constituents, Quantitative Analysis and Antioxidant Activities of Echinacea purpurea (L.) Moench and Echinacea pallida (Nutt.) Nutt. Journal of Food Biochemistry, 39(5), 622-630. https://doi.org/10.1111/jfbc.12168
• Erenler, R., Uno, M., Goud, T. V., & Biellmann, J. F. (2009). Preparation of some heterocyclic enones and ynones by isomerisation of the propargylic alcohols. Journal of Chemical Research, 2009(7), 459-464. https://doi.org/10.3184/030823409X464467
• Erenler, R., Yaman, C., Demirtas, I., & Alma, M. H. (2023). Phytochemical Investigation of Hypericum heterophyllum Flowers: LC-ESI-MS/MS Analysis, Total Phenolic and Flavonoid Contents, Antioxidant Activity. The Natural Products Journal, 13(7), e120123212672. https://doi.org/10.2174/2210315513666230112165545
• Erenler, R., Yilmaz, S., Aksit, H., Sen, O., Genc, N., Elmastas, M., & Demirtas, I. (2014). Antioxidant activities of chemical constituents isolated from Echinops orientalis Trauv. Records of Natural Products, 8(1), 32-36.
• Ganeshpurkar, A., & Saluja, A. K. (2017). The Pharmacological Potential of Rutin. Saudi Pharmaceutical Journal, 25(2), 149-164. https://doi.org/10.1016/j.jsps.2016.04.025
• Gecer, E. N. (2021). Green Synthesis of Silver Nanoparticles from Salvia aethiopis L. and Their Antioxidant Activity. Journal of Inorganic and Organometallic Polymers and Materials, 31(11), 4402-4409. https://doi.org/10.1007/s10904-021-02057-3
• Gecer, E. N. (2023). Synthesis and characterization of silver nanoparticles using Origanum onites leaves: Cytotoxic, apoptotic, and necrotic effects on Capan-1, L929, and Caco-2 cell lines. Green Processing and Synthesis, 12(1), 20228126. https://doi.org/10.1515/gps-2022-8126
• Gecer, E. N., & Erenler, R. (2022a). Biosynthesis of silver nanoparticles using Dittrichia graveolens (Asteraceae) leaves extract: characterisation and assessment of their antioxidant activity. Turkish Journal of Biodiversity, 5(1), 50-56. https://doi.org/10.38059/biodiversity.1090549
• Gecer, E. N., & Erenler, R. (2022b). Biosynthesis of Silver Nanoparticles using Echinacea pallida (Nutt.) Nutt. and Antioxidant Activity Thereof. Journal of the Chemical Society of Pakistan, 44(6), 610-617. https://doi.org/10.52568/001187/JCSP/44.06.2022
• Gecer, E. N., & Erenler, R. (2023). Biogenic synthesis of silver nanoparticles using Echium vulgare: Characterisation, quantitative analysis of bioactive compounds, antioxidant activity and catalytic degradation. Journal of the Indian Chemical Society, 100(5), 101003. https://doi.org/10.1016/j.jics.2023.101003
• Gecer, E. N., Erenler, R., Temiz, C., Genc, N., & Yildiz, I. (2022). Green synthesis of silver nanoparticles from Echinacea purpurea (L.) Moench with antioxidant profile. Particulate Science and Technology, 40(1), 50-57. https://doi.org/10.1080/02726351.2021.1904309
• Genc, N., Yildiz, I., Chaoui, R., Erenler, R., Temiz, C., & Elmastas, M. (2021). Biosynthesis, characterization and antioxidant activity of oleuropein-mediated silver nanoparticles. Inorganic and Nano-Metal Chemistry, 51(3), 411-419. https://doi.org/10.1080/24701556.2020.1792495
• Genç, N., Elmastaş, M., Telci, İ., & Erenler, R. (2020). Quantitative analysis of phenolic compounds of commercial basil cultivars (Ocimum basilicum L.) by LC-TOF-MS and their antioxidant effects. International Journal of Chemistry and Technology, 4(2), 179-184. http://dx.doi.org/10.32571/ijct.795629
• Guemidi, C., Ait Saada, D., Ait Chabane, O., Elmastas, M., Erenler, R., Yilmaz, M. A., Tarhan, A., Akkal, S., & Khelifi, H. (2024). Enhancement of yogurt functionality by adding Mentha piprita phenolic extract and evaluation of its quality during cold storage. Food Science & Nutrition, 12, 3007-3020. https://doi.org/10.1002/fsn3.3981
• Hadjra, H., Yamina, B., Soulef, K., & Erenler, R. (2023). Evaluation of Antifungal Potential of Mentha pulegium Essential oil in Biological Control Against the Pathogen of Inflorescence Rot Disease of Date Palm (Mauginiella scaettae). Jordan Journal of Biological Sciences, 16(4), 665-672. https://doi.org/10.54319/jjbs/160412
• Houari, F. Z., Brahmi, M., Erenler, R., & Hariri, A. (2024). Chemical profile, antibacterial and antioxidant properties of Rubia tinctorum L. essential oils. Natural Product Research, 165-174. https://doi.org/10.1080/14786419.2024.2356670
• Kaya, G., Karakaya, R., Tilgel, E., Sandikci, M., Yucel, E., Cicek, G., Kayir, O., Aksit, H., Telci, I., & Guzel, A. (2014). Essential Oil Constituents of Thuja orientalis Berries. Journal of New Results in Science, 7(7), 1-6
• Khodja, E. A. T., Abd El Hamid Khabtane, R. A., Benouchenne, D., Bensaad, M. S., Bensouici, C., & Erenler, R. (2023). In vitro assessment of antioxidant, neuroprotective, anti-urease and anti-tyrosinase capacities of Tamarix africana leaves extracts. Journal of Traditional Chinese Medicine, 43(2), 252. https://doi.org/10.19852/j.cnki.jtcm.20230105.003
• Kolodziejczyk-Czepas, J. (2012). Trifolium species-derived substances and extracts—Biological activity and prospects for medicinal applications. Journal of Ethnopharmacology, 143(1), 14-23. https://doi.org/10.1016/j.jep.2012.06.048
• Lu, J., Maezawa, I., Weerasekara, S., Erenler, R., Nguyen, T. D. T., Nguyen, J., Swisher, L. Z., Li, J., Jin, L. W., Ranjan, A., Srivastava, S. K., & Hua, D. H. (2014). Syntheses, neural protective activities, and inhibition of glycogen synthase kinase-3β of substituted quinolines. Bioorganic and Medicinal Chemistry Letters, 24(15), 3392-3397. https://doi.org/10.1016/j.bmcl.2014.05.085
• McKenna, P., Cannon, N., Conway, J., & Dooley, J. (2018). The use of red clover (Trifolium pratense) in soil fertility-building: A Review. Field Crops Research, 221, 38-49. https://doi.org/10.1016/j.fcr.2018.02.006
• Ortiz, A. d. C., Fideles, S. O. M., Reis, C. H. B., Bellini, M. Z., Pereira, E. d. S. B. M., Pilon, J. P. G., de Marchi, M. Â., Detregiachi, C. R. P., Flato, U. A. P., & Trazzi, B. F. d. M. (2022). Therapeutic effects of citrus flavonoids neohesperidin, hesperidin and its aglycone, hesperetin on bone health. Biomolecules, 12(5), 626. https://doi.org/10.3390/biom12050626
• Randjelović, P., Veljković, S., Stojiljković, N., Sokolović, D., Ilić, I., Laketić, D., Randjelović, D., & Randjelović, N. (2015). The beneficial biological properties of salicylic acid. Acta facultatis medicae Naissensis, 32(4), 259-265. https://doi.org/10.1515/afmnai-2015-0026
• Sahin Yaglioglu, A., Akdulum, B., Erenler, R., Demirtas, I., Telci, I., & Tekin, S. (2013). Antiproliferative activity of pentadeca-(8E, 13Z) dien-11-yn-2-one and (E)-1,8-pentadecadiene from Echinacea pallida (Nutt.) Nutt. roots. Medicinal Chemistry Research, 22(6), 2946-2953. https://doi.org/10.1007/s00044-012-0297-2
• Sahin Yaglioglu, A., Erenler, R., Gecer, E. N., & Genc, N. (2022). Biosynthesis of Silver Nanoparticles Using Astragalus flavesces Leaf: Identification, Antioxidant Activity, and Catalytic Degradation of Methylene Blue. Journal of Inorganic and Organometallic Polymers and Materials, 32, 3700-3707. https://doi.org/10.1007/s10904-022-02362-5
• Topçu, G., Erenler, R., Çakmak, O., Johansson, C. B., Çelik, C., Chai, H.-B., & Pezzuto, J. M. (1999). Diterpenes from the berries of Juniperus excelsa. Phytochemistry, 50(7), 1195-1199. https://doi.org/10.1016/S0031-9422(98)00675-X
• Türkmen, N., Öz, A., Sönmez, A., Erol, T., Gülümser, D., Yurdakul, B., Kayır, Ö., Elmastas, M., & Erenler, R. (2014). Chemical Composition of Essential Oil from Rosmarinus Officinalis L. Leaves. Journal of New Results in Science, 6(6), 27-31.
• Yaman, C., Önlü, Ş., Ahmed, H., & Erenler, R. (2022). Comparison of phytochemicals and antioxidant capacity of Hypericum perforatum; wild plant parts and in vitro samples. Journal of Animal & Plant Sciences, 32(2), 596-603. https://doi.org/10.36899/JAPS.2022.2.0459
• Zaoui, H., Boutaoui, N., Menad, A., Erenler, R., Zaiter, L., Fadilab, B., & Souada, A. (2022). Phytochemical Profile, Anti-lipid peroxidation and Anti-diabetic activities of Thymus algeriensis Boiss. & Reut. Egyptian Journal of Chemistry, 65(12), 695-705.
• Zerrouki, S., Mezhoud, S., Sahin Yaglioglu, A., Bensouici, C., Nuri Atalar, M., Demirtas, I., Ameddah, S., & Mekkiou, R. (2022). Antioxidant, anticancer activities, and HPLC-DAD analyses of the medicinal halophyte Limoniastrum guyonianum Dur. extracts. Journal of Research in Pharmacy, 26(3), 598-608. https://doi.org/10.29228/jrp.157