Quantitative analysis and antioxidant activity of Trifolium pratense L. stem

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

doi:10.53445/batd.1669871

TR EN

Quantitative analysis and antioxidant activity of Trifolium pratense L. stem

Abstract

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.

Keywords

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

References

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

Details

Primary Language

English

Subjects

Chemical Engineering (Other), Naturopathy

Journal Section

Research Article

Authors

Ramazan Erenler ^*
0000-0002-0505-3190
Türkiye

İlyas Yıldız
0000-0003-1254-1069
Türkiye

Esma Nur Geçer
0000-0002-0095-079X
Türkiye

İbrahim Hosaflıoğlu
0000-0002-0455-0515
Türkiye

Süleyman Muhammed Çelik
0000-0001-6025-8298
Türkiye

Early Pub Date

April 30, 2025

Publication Date

April 30, 2025

Submission Date

April 4, 2025

Acceptance Date

April 26, 2025

Published in Issue

Year 2025 Volume: 6 Number: 1

DOI

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

IZ

https://izlik.org/JA65DX77DW

APA

Erenler, R., Yıldız, İ., Geçer, E. N., Hosaflıoğlu, İ., & Çelik, S. M. (2025). Quantitative analysis and antioxidant activity of Trifolium pratense L. stem. Journal of Integrative and Anatolian Medicine, 6(1), 1-8. https://doi.org/10.53445/batd.1669871

AMA

1.Erenler R, Yıldız İ, Geçer EN, Hosaflıoğlu İ, Çelik SM. Quantitative analysis and antioxidant activity of Trifolium pratense L. stem. J. Integr. Anatol. Med. 2025;6(1):1-8. doi:10.53445/batd.1669871

Chicago

Erenler, Ramazan, İlyas Yıldız, Esma Nur Geçer, İbrahim Hosaflıoğlu, and Süleyman Muhammed Çelik. 2025. “Quantitative Analysis and Antioxidant Activity of Trifolium Pratense L. Stem”. Journal of Integrative and Anatolian Medicine 6 (1): 1-8. https://doi.org/10.53445/batd.1669871.

EndNote

Erenler R, Yıldız İ, Geçer EN, Hosaflıoğlu İ, Çelik SM (April 1, 2025) Quantitative analysis and antioxidant activity of Trifolium pratense L. stem. Journal of Integrative and Anatolian Medicine 6 1 1–8.

IEEE

[1]R. Erenler, İ. Yıldız, E. N. Geçer, İ. Hosaflıoğlu, and S. M. Çelik, “Quantitative analysis and antioxidant activity of Trifolium pratense L. stem”, J. Integr. Anatol. Med., vol. 6, no. 1, pp. 1–8, Apr. 2025, doi: 10.53445/batd.1669871.

ISNAD

Erenler, Ramazan - Yıldız, İlyas - Geçer, Esma Nur - Hosaflıoğlu, İbrahim - Çelik, Süleyman Muhammed. “Quantitative Analysis and Antioxidant Activity of Trifolium Pratense L. Stem”. Journal of Integrative and Anatolian Medicine 6/1 (April 1, 2025): 1-8. https://doi.org/10.53445/batd.1669871.

JAMA

1.Erenler R, Yıldız İ, Geçer EN, Hosaflıoğlu İ, Çelik SM. Quantitative analysis and antioxidant activity of Trifolium pratense L. stem. J. Integr. Anatol. Med. 2025;6:1–8.

MLA

Erenler, Ramazan, et al. “Quantitative Analysis and Antioxidant Activity of Trifolium Pratense L. Stem”. Journal of Integrative and Anatolian Medicine, vol. 6, no. 1, Apr. 2025, pp. 1-8, doi:10.53445/batd.1669871.

Vancouver

1.Ramazan Erenler, İlyas Yıldız, Esma Nur Geçer, İbrahim Hosaflıoğlu, Süleyman Muhammed Çelik. Quantitative analysis and antioxidant activity of Trifolium pratense L. stem. J. Integr. Anatol. Med. 2025 Apr. 1;6(1):1-8. doi:10.53445/batd.1669871