Rumex Crispus L.’dan Sekonder Metabolitlerin Kromatografik Yöntemlerle İzolasyonu ve Tayini

Yıl 2024, Cilt: 24 Sayı: 06, 1305 - 1312

Elif Günay , Dilek Mutlu , Deniz Küpçük , Gizem Yıldırım Baştemur , Sabriye Perçin Özkorucuklu

Öz

İnsanlar binlerce yıldır bitkilerin sağlık açısından faydalı özelliklerinden yararlanmaktadır. Günümüzde, tıp biliminin yanı sıra başta kozmetik endüstrisinde olmak üzere birçok sektörde bitkiler kullanılmaktadır ve bitkilerin fitokimyasal içeriklerinin belirlenmesine ilişkin çalışmalar da hızla artmaktadır. Halk arasında çeşitli amaçlarla kullanılan Rumex crispus L. bitkisi özellikle fenolik bileşikler ve antrakinonlar olmak üzere zengin sekonder metabolit içeriğinden dolayı dikkat çekmektedir. Bu çalışmada Türkiye’de yetişen Rumex crispus L. bitkisinden katma değere sahip fenolik bileşik ve antrakinonların izolasyonu pratik ve ekonomik bir metot olan gravity kolon kromatografisi ile gerçekleştirilmiştir. Ekstrakt apolar çözücü sistemden polar çözücü sistemine doğru fraksiyonlara ayrılmış ve bu fraksiyonlardaki fenolik bileşik ve antrakinon miktarları ters faz sıvı kromatografi (RPLC) yöntemi ile belirlenmiştir. Protokateşik asit, gentisik asit, kafeik asit, p-kumarik asit, polydatin, kumarin, resveratrol ve ellajik asit izolasyonu için etil asetat-metanol (6:4) çözücü sisteminin uygun olduğuna karar verilmiştir. Sonuç olarak, bitkilerdeki sekonder metabolitlerin araştırılmasında maliyeti düşük olan gravity kolon kromatografisi kullanılarak fenolik bileşiklerin ve antrakinonların saflaştırılabileceği ortaya konulmuştur.

Anahtar Kelimeler

Rumex crispus L., Gravity kolon kromatografi, RPLC, Fenolik bileşik, Antrakinon.

Etik Beyan

Yazarlar tüm etik standartlara uyduklarını beyan ederler.

Destekleyen Kurum

İstanbul Üniversitesi Bilimsel Araştırma Projeleri Koordinasyon Birimi

Proje Numarası

FLO-2023-40354

Teşekkür

Bu çalışma İstanbul Üniversitesi Bilimsel Araştırma Projeleri Koordinasyon Birimi tarafından desteklenmiştir (Proje numarası: FLO-2023-40354). İstanbul Üniversitesi Fen Fakültesi Botanik Anabilim Dalından Doç. Dr. İbrahim Sırrı Yüzbaşıoğlu’na bitki tür tayinindeki katkılarından dolayı teşekkür ederiz.

Isolation and Determination of Secondary Metabolites from Rumex Crispus L. by Chromatographic Methods

Öz

People have been benefiting from the health benefits of plants for thousands of years. Nowadays, plants are used in many sectors, especially in the cosmetic industry, as well as medical science, and studies on the determination of the phytochemical contents of plants are rapidly increasing. Rumex crispus L., a plant commonly used for various purposes by the public, attracts attention due to its rich secondary metabolite content, especially phenolic compounds and anthraquinones. In this study, the isolation of valuable phenolic compounds and anthraquinones from the Rumex crispus L. plant grown in Turkey was carried out by gravity column chromatography, which is a practical and economical method. The extract was divided into fractions from the nonpolar solvent system to the polar solvent system, and the amounts of phenolic compounds and anthraquinones in these fractions were determined by the reverse phase liquid chromatography (RPLC) method. It was decided that the ethyl acetate-methanol (6:4) solvent system was suitable for the isolation of protocatechuic acid, gentisic acid, caffeic acid, p-coumaric acid, polydatin, coumarin, resveratrol and ellagic acid. Consequently, it has been revealed that phenolic compounds and anthraquinones can be purified by using low-cost gravity column chromatography in the investigation of secondary metabolites in plants.

Anahtar Kelimeler

Rumex crispus L., Gravity column chromatography, RPLC, Phenolic compound, Anthraquinone.

Proje Numarası

FLO-2023-40354

Kaynakça

• Abdisa Z. ve Zelalem G. (2022). Phytochemical Investigation of Rumex Abyssinicus Root Barks and In Vitro Evaluation of Its Antibacterial Activities. Journal of the Turkish Chemical Society Section A Chemistry, 9(4), 1063-1072. https://doi.org/10.18596/jotcsa.1035039
• Arora, M., Mahajan, A., ve Sembi, J. K. (2022). Extraction and characterization of secondary metabolites from Crepidium acuminatum (D. Don) Szlach: A miraculous orchid. South African Journal of Botany, 149, 693-700. https://doi.org/10.1016/j.sajb.2022.06.014
• Akpınar, R., Yıldırım Baştemur, G., Bıçak, B., Sanli, N. O., Mertoğlu Kamalı, E., Pekmez, M., Kecel Gündüz, S., ve Perçin Özkorucuklu, S. (2024). Phytochemical profiling, in vitro biological activities, and in silico (molecular docking and absorption, distribution, metabolism, excretion, toxicity) studies of Polygonum cognatum Meissn. Journal of Separation Science, 47(1), 2300750. https://doi.org/10.1002/jssc.202300750
• Amarowicz, R., Shahiiİ, F., ve Wiczkowski, W. (2003). Separation of indıvidual catechins from green tea using silica gel column chromatography and HPLC. Journal of Food Lipids, 10(2), 165–177. https://doi.org/10.1111/j.1745-4522.2003.tb00013.x
• Aybastıer, Ö., (2021). Karaduttan (Morus Nigra L.) antioksidan bileşiklerin kromatografik yöntemlerle izolasyonu ve tayini. Gıda, 46 (1), 32-41. https://doi.org/10.15237/gida.GD20114
• Bajpai, V. K., Majumder, R., ve Park, J. G. (2016). Isolation and purification of plant secondary metabolites using column-chromatographic technique. Bangladesh Journal of Pharmacology, 11(4), 844-848. https://doi.org/10.3329/bjp.v11i4.28185
• Barba, F. J., Esteve, M. J., ve Frígola, A. (2014). Bioactive Components from Leaf VegetableProducts, Studies in Natural Products Chemistry, In: Atta ur, R. (ed.), Chapter 11, Elsevier, ISBN: 1572-5995, 321-346.
• Başkan, S., Daut-Ozdemir, A., Günaydin, K., ve Erim, F. B. (2007). Analysis of anthraquinones in Rumex crispus by micellar electrokinetic chromatography. Talanta, 71(2), 747–750. https://doi.org/10.1016/j.talanta.2006.05.018
• Bulduk İ. ve Gökçe S., (2021). Farmasötik Formülasyonlarda Levodopa ve Karbidopa Miktarının Belirlenmesi için Etkili ve Güvenilir bir HPLC Yönteminin Geliştirilmesi ve Validasyonu. Hacettepe Journal of Biology and Chemistry, 49 (4), 413-422. https://doi.org/10.15671/hjbc.773988
• Çoruh, I., Gormez, A., Ercisli, S., ve Sengul, M. (2008). Total phenolic content, antioxidant, and antibacterial activity of Rumex crispus grown wild in Turkey. Pharmaceutical Biology, 46(9), 634-638. https://doi.org/10.1080/13880200802182240
• Duval, J., Pecher, V., Poujol, M., ve Lesellier, E. (2016). Research advances for the extraction, analysis and uses of anthraquinones: A review. Industrial Crops and Products, 94, 812-833. https://doi.org/10.1016/j.indcrop.2016.09.056
• Eom T, Kim E, ve Kim J.S. (2020), In vitro antioxidant, antiinflammation, and anticancer activities and anthraquinone content from Rumex crispus root extract and fractions. Antioxidants (Basel), 9(726), 1-13. https://doi.org/10.3390/antiox9080726
• Fan, M., Ding, H., Zhang, G., Hu, X., ve Gong, D. (2019). Relationships of dietary flavonoid structure with its tyrosinase inhibitory activity and affinity. LWT - Food Science and Technology, 107, 25-34. https://doi.org/10.1016/j.lwt.2019.02.076
• Fouillaud, M., Venkatachalam, M., Girard-Valenciennes,E., Caro, Y., ve Dufossé, L. (2016). Anthraquinones and derivatives from marine-derived fungi: Structural diversity and selected biological activities. Marine Drugs, 14(4), 64. https://doi.org/10.3390/md14040064
• Guo, S., Feng, B., Zhu, R., Ma, J., ve Wang, W. (2011). Preparative isolation of three anthraquinones from Rumex japonicus by high-speed counter-current chromatography. Molecules, 16(2), 1201–1210. https://doi.org/10.3390/molecules16021201
• Huang, P-H., Huang, C-Y., Chen, M-C., Lee, Y-T., Yue, C-H., Wang, H-Y., ve Lin, H. (2013). Emodin and aloe-emodin suppress breast cancer cell proliferation through ER alpha Inhibition. Evidence-based Complementary and Alternative Medicine, 376123. https://doi.org/10.1155/2013/376123
• Idris, O. A., Wintola, O. A., ve Afolayan, A. J. (2017). Phytochemical and antioxidant activities of Rumex crispus L. in treatment of gastrointestinal helminths in Eastern Cape Province, South Africa. Asian Pacific Journal of Tropical Biomedicine, 7(12), 1071–1078. https://doi.org/10.1016/j.apjtb.2017.10.008
• Larayetan, R., Ojemaye, M.O., Okoh, O.O., ve Okoh, A.I. (2019). Silver nanoparticles mediated by Callistemon citrinus extracts and their antimalaria, antitrypanosoma and antibacterial efficacy. Journal of Molecular Liquids, 273, 615-625. https://doi.org/10.1016/j.molliq.2018.10.020
• Li-Weber, M. (2013). Targeting apoptosis pathways in cancer by Chinese medicine. Cancer Letters, 332(2), 304-312. https://doi.org/10.1016/j.canlet.2010.07.015
• Minh, T. N., Van, T. M., Andriana, Y., Vinh, L. T., Hau, D. V., Duyen, D. H., ve Guzman-Gelani, C. D. (2019). Antioxidant, xanthine oxidase, α-amylase and α-glucosidase inhibitory activities of bioactive compounds from Rumex crispus L. root. Molecules, 24(21), 3899. https://doi.org/10.3390/molecules24213899
• Nazeam, J. A., Al-Shareef, W. A., Helmy, M. W., & El-Haddad, A. E. (2020). Bioassay-guided isolation of potential bioactive constituents from pomegranate agrifood by-product. Food Chemistry, 326, 126993. https://doi.org/10.1016/j.foodchem.2020.126993
• Nuzula, A. F., Kristanti, A. N., Aminah, N. S., Takaya, Y., Sahadatin, M. N., ve Wardana, A. P. (2023). Isolation and structure elucidation of secondary metabolite compounds from Curcuma aeruginosa. Jurnal Kimia Riset, 8(1), 81–91. https://doi.org/10.20473/jkr.v8i1.44073
• Ntemafack, A., Sabeena, A., Aristide, D., Ravi, S., Serge, A., Jules-Roger, K., Alain, W., Asha, Chaubey. (2023). Chemical profile, biological potential, bioprospection and biotechnological application of endophytes of Rumex: A systematic review. Industrial Crops & Products, 195, 116474. https://doi.org/10.1016/j.indcrop.2023.116474
• Orban-Gyapai, O., Liktor-Busa, E., Kusz, N., Stefko, D., Urban, E., Hohmann, J., ve Vasas, A. (2017). Antibacterial screening of Rumex species native to the Carpathian Basin and bioactivity-guided isolation of compounds from Rumex aquaticus. Fitoterapia, 118, 101-106. https://doi.org/10.1016/j.fitote.2017.03.009
• Pham, D. Q., Han, J. W., Dao, N. T., Kim, J.-C., Pham, H. T., Nguyen, T. H., Choib G.J., Vu H.D., Le Dang, Q. (2020). In vitro and in vivo antimicrobial potential against various phytopathogens and chemical constituents of the aerial part of Rumex chinensis Campd. South African Journal of Botany, 133, 73–82. https://doi.org/10.1016/j.sajb.2020.07.006
• Saoudi, M. M., Bouajila, J., Rahmani, R., ve Alouani, K. (2021). Phytochemical Composition, Antioxidant, Antiacetylcholinesterase, and Cytotoxic Activities of Rumex crispus L. International Journal of Analytical Chemistry, 6675436. https://doi.org/10.1155/2021/6675436
• Sarikahya, N. B., Goren, A. C., ve Kirmizigul, S. (2019). Simultaneous determination of several flavonoids and phenolic compounds in nineteen different Cephalaria species by HPLC-MS/MS. Journal of Pharmaceutical and Biomedical Analysis, 173, 120-125. https://doi.org/10.1016/j.jpba.2019.05.019
• Seigler, D. S. (2012). Plant secondary metabolism. Springer Science & Business Media.
• Simoes, C.M.O., Schenkel, E.P., Gosman, G., Mello, J.C.P., Mentz, L.A., ve Petrovick, P.R. (2007). Farmacognosia: da planta ao medicamento. Editora da UFSC, Floriano´polis, 1102.
• Skoog, D. A., West, D. M., Holler, F. J., & Crouch, S. R. (2013). Fundamentals of Analytical Chemistry. Cengage learning.
• Smolarz, H.D., Swatko-Ossor, M., Ginalska, G., ve Medynska, E. (2013). Antimycobacterial effect of extract and its components from Rheum rhaponticum. Journal of AOAC International, 96(1), 155-160. https://doi.org/10.5740/jaoacint.12-010
• Tsamo, L. D. F., Yimgang, L. V., Wouamba, S. C. N., Mkounga, P., Nkengfack, A. E., Voutquenne-Nazabadioko, L., ve Sewald, N. (2021). A new ceramide (Rumexamide) and other chemical constituents from Rumex abyssinicus Jacq (Polygonaceae): isolation, characterization, antibacterial activities and chemophenetic significance. Advances in Biological Chemistry, 11(05), 266-282. https://doi.org/10.4236/abc.2021.115018
• Vasas, A., Orbán-Gyapai, O., ve Hohmann, J. (2015). The genus Rumex: review of traditional uses, phytochemistry, and pharmacology. Journal of Ethnopharmacology, 175, 198-228. https://doi.org/10.1016/j.jep.2015.09.001
• WHO. (1993) Summar WHO guidelines for the assessment of herbal medicines. Herbal Grom. 28:13–4.
• Yıldırım Baştemur, G., Akpınar, R., Kır, E., ve Perçin Özkorucuklu, S., (2024). Development and Validation of the HPLC-DAD Method for the Simultaneous Determination of Anthraquinones in Rumex crispus L. and Rumex acetosella L. Plants and Evaluation of Their Antioxidant Capacities. Journal of Analytical Chemistry,79(2), 179-189. https://doi.org/10.1134/S1061934824020035
• Yıldırım, A., Mavi, A., ve Kara, A.A. (2001). Determination of Antioxidant and Antimicrobial Activities of Rumex crispus L. Extracts. Journal of Agricultural and Food Chemistry, 49, 4083–4089. https://doi.org/10.1021/jf0103572