Cephalaria tchihatchewii Boiss. Ekstrelerinin Antioksidan Aktivitesi, Metabolik Enzimler Üzerine Etkisi ve UPLC-MS/MS Analizi ile Kimyasal Karakterizasyonun Belirlenmesi

Abstract

Bu çalışma, Cephalaria tchihatchewii Boiss'in metanol ve su ekstrelerinin (MECT, WECT) UPLC-MS/MS kullanarak kapsamlı bir kimyasal karakterizasyonunu gerçekleştirmeyi ve bu ekstrelerin potansiyel antioksidan aktivitesinin ve bazı metabolik enzimler üzerindeki etkisini değerlendirmeyi amaçlamaktadır. Antioksidan aktivite, DPPH, ABTS, DMPD, FRAP, KUPRAK ve Fe3+ indirgeme testleri dahil olmak üzere çeşitli testler kullanılarak değerlendirilmiştir. MECT ve WECT'in enzim inhibisyon IC50 değerleri, sırasıyla α-glikozidaz için 28.98 ve 34.19 μg/mL, asetilkolinesteraz için 21.82 ve 26.52 μg/mL ve karbonik anhidraz II için 27.75 ve 13.72 μg/mL’dir. MECT ve WECT'de bulunan fenolik bileşik miktarları sırasıyla 47.00 ve 46.00 μg GAE /mg ekstre, flavonoit miktarları 60.98 ve 49.34 μg KE/mg ekstre olarak belirlenmiştir. MECT ve WECT’in IC50 değerleri sırasıyla DPPH için 34.66 ve 57.76 μg/mL, ABTS için 20.39 ve 17.33 μg/mL ve DMPD için 53.32 ve 57.76 g/mL’dir. Ekstrelerin indirgeme yetenekleri ise sırasıyla Fe+3 indirgeme (λ700:0.377 ve 0.680), FRAP (λ593:0.690 ve 0.369) ve KUPRAK (λ450:0.458 ve 0.333)’dür. UPLC-MS/MS analiziyle elde edilen sonuçlara göre ana bileşenler MECT’in kinik asit 0.39844 µg/mL, klorojenik asit 38.412 µg/mL, fumarik asit 0.301 µg/mL ve WECT’in klorojenik asit 13.639 µg/mL, kinik asit 11.004 µg/mL, siyanidin-3-O-glukozit 0.778 µg/mL olduğu belirlenmiştir. Ekstrelerin, antioksidan aktivite ve α-glikosidaz, asetilkolinesteraz, karbonik anhidraz II enzim ihbisyon etkisi dahil olmak üzere çeşitli biyolojik aktivitelere sahip olduğu belirlenmiştir.

Keywords

• Karbonik anhidraz II
• Kolinesteraz
• α-Glikozidaz
• Antioksidan
• Enzim inhibisyon
• UPLC-MS/MS
• Cephalaria

Determination of Cephalaria tchihatchewii Boiss.'s Extracts Antioxidant Activity, Effect on Metabolic Enzymes, and Chemical Characterization by UPLC-MS/MS Analysis

Abstract

The present study aims to perform a comprehensive chemical characterization of the methanol and water extracts of Cephalaria tchihatchewii Boiss. (MECT, WECT) using UPLC-MS/MS, besides evaluating the potential antioxidant activity of these extracts and their effect on some metabolic enzymes. The antioxidant activity was assessed using various assays including DPPH, ABTS, DMPD, FRAP, CUPRAC, and Fe3+ reducing assays. The inhibition effects of the extracts against α-glycosidase, acetylcholinesterase and carbonic anhydrase II enzymes were evaluated. The IC50 values of MECT and WECT were for α-glycosidase were 28.98 and 34.19 μg/mL, for acetylcholinesterase were 21.82 and 26.52 μg/mL, and for carbonic anhydrase, 27.75 and 13.72 μg/mL, respectively. The amounts of phenolic compounds present in MECT and WECT were determined to be 47.00 and 46.00 μg GAE /mg extract, respectively. The amounts of flavonoits present in MECT and WECT were determined to be 60.98 and 49.34 μg KE/mg extract, respectively. The MECT and WECT exhibited distinct activities on DPPH, ABTS, and DMPD. The IC50 values were for DPPH 34.66 and 57.76 μg/mL for ABTS 20.39 and 17.33 µg/mL and for DMPD 53.32 and 57.76 g/mL, respectively. Reducing abilities of extracts were respectively Fe+3 reducing (λ700:0.377 and 0.680), FRAP (λ593:0.690 and 0.369), and CUPRAC (λ450:0.458 and 0.333). UPLC-MS/MS revealed that the major components are MECT (quinic acid 39.844 µg/mL, chlorogenic acid 38.412 µg/mL, fumaric acid 0.301 µg/mL), and WECT (chlorogenic acid 13.639 µg/mL, qunic acid 11.004 µg/mL, cyanidin-3-O-glucoside 0.778 µg/mL). The extracts were determined to have various biological activities, including antioxidant activity and α-glycosidase, acetylcholinesterase, carbonic anhydrase II enzyme inhibition effect.

Keywords

• Carbonic anhydrase II
• Cholinesterase
• α-Glycosidase
• Enzyme inhibition
• Antioxidant
• UPLC-MS/MS
• Cephalaria

References

1. Ak, T., & Gülçin, İ. (2008). Antioxidant and radical scavenging properties of curcumin. Chemico-Biological Interactions, 174(1), 27-37. https://doi.org/10.1016/j.cbi.2008.05.003
2. Alam, M. N., Bristi, N. J., & Rafiquzzaman, M. (2013). Review on in vivo and in vitro methods evaluation of antioxidant activity. Saudi pharmaceutical journal, 21(2), 143-152.
3. Apak, R., Güçlü, K., Özyürek, M., Esin Karademir, S., & Erçağ, E. (2006). The cupric ion reducing antioxidant capacity and polyphenolic content of some herbal teas. International Journal of Food Sciences and Nutrition, 57(5-6), 292-304. https://doi.org/10.1080/09637480600798132
4. Arabaci, B., Gulcin, I., & Alwasel, S. (2014). Capsaicin: A Potent Inhibitor of Carbonic Anhydrase Isoenzymes. Molecules, 19(7), 10103-10114.
5. Atalan, E. (2019). Türkiye’de yetiştirilen pelemir bitkisinin [Cephalaria syriaca (l.)] antioksidan, antimikrobiyal, antifungal, antibiyofilm özelliklerinin ve tohum morfolojisinin araştırılması. Bartın Üniversitesi, Fen Bilimleri Enstitüsü.
6. Blois, M. S. (1958). Antioxidant Determinations by the Use of a Stable Free Radical. Nature, 181(4617), 1199-1200. https://doi.org/10.1038/1811199a0
7. Burak, B., Emel, M. K., & Yeter, Y. C. (2021). Total phenolic, total flavonoit contents, and in vitro biological activities of Cephalaria procera. İstanbul Journal of Pharmacy, 51(3), 365-371.
8. Celenk, V. U., Sarikahya, N. B., & Kirmizigul, S. (2020). Isolation and Structural Studies on Saponins from Three Cephalaria Species from Anatolia. Chemistry of Natural Compounds, 56(1), 180-182.

9. Chrząszcz, M., Miazga-Karska, M., Klimek, K., Granica, S., Tchórzewska, D., Ginalska, G., & Szewczyk, K. (2020a). Extracts from Cephalaria Uralensis (Murray) Roem. & Schult. and Cephalaria Gigantea (Ledeb.) Bobrov as Potential Agents for Treatment of Acne Vulgaris: Chemical Characterization and In Vitro Biological Evaluation. Antioxidants, 9, 796. doi:10.3390/antiox9090796
10. Chrząszcz, M., Miazga-Karska, M., Klimek, K., Granica, S., Tchórzewska, D., Ginalska, G., & Szewczyk, K. (2020b). Extracts from Cephalaria Uralensis (Murray) Roem. & Schult. and Cephalaria Gigantea (Ledeb.) Bobrov as Potential Agents for Treatment of Acne Vulgaris: Chemical Characterization and In Vitro Biological Evaluation. Antioxidants, 9(9), 796.
11. Dagli, M., & Sarıkahya, N. (2018). Elucidation and characterization of glycosidic compounds from Cephalaria tuteliana (dipsacaceae). (master of science thesis), Ege University, Bornova-IZMIR.
12. Dagli, M., Sarikahya, N. B., Nalbantsoy, A., & Kirmizigul, S. (2019). Comparative Phytochemical Screening and Cytotoxic Efficacy of Endemic Cephalaria tuteliana. Natural Product Communications, 14(10), 1934578X19862647.
13. Durmaz, L., Kiziltas, H., Guven, L., Karagecili, H., Alwasel, S., & Gulcin, İ. (2022). Antioxidant, Antidiabetic, Anticholinergic, and Antiglaucoma Effects of Magnofluorine. Molecules, 27(18), 5902.
14. Ellman, G. L., Courtney, K. D., Andres, V., & Featherstone, R. M. (1961). A new and rapid colorimetric determination of acetylcholinesterase activity. Biochemical Pharmacology, 7(2), 88-95. https://doi.org/10.1016/0006-2952(61)90145-9
15. Erdogan, M. K., Gundogdu, R., Yapar, Y., Gecibesler, I. H., Kirici, M., Behcet, L., . . . Taslimi, P. (2023). In vitro anticancer, antioxidant and enzyme inhibitory potentials of endemic Cephalaria elazigensis var. purpurea with in silico studies. Journal of Biomolecular Structure and Dynamics, 1-13. doi:10.1080/07391102.2022.2163700
16. Eruygur, N., Koçyiğit, U. M., Taslimi, P., Ataş, M., Tekin, M., & Gülçin, İ. (2019). Screening the in vitro antioxidant, antimicrobial, anticholinesterase, antidiabetic activities of endemic Achillea cucullata (Asteraceae) ethanol extract. South African Journal of Botany, 120, 141-145. doi:https://doi.org/10.1016/j.sajb.2018.04.001
17. Fogliano, V., Verde, V., Randazzo, G., & Ritieni, A. (1999). Method for measuring antioxidant activity and its application to monitoring the antioxidant capacity of wines. Journal of agricultural and food chemistry, 47(3), 1035-1040.
18. Göçer, H., & Gülçin, İ. (2011). Caffeic acid phenethyl ester (CAPE): correlation of structure and antioxidant properties. International Journal of Food Sciences and Nutrition, 62(8), 821-825. doi:10.3109/09637486.2011.585963
19. Göktürk, R. S., & Sümbül, H. (2014). A taxonomic revision of the genus Cephalaria (Caprifoliaceae) in Turkey. Turkish Journal of Botany, 38, 927-968. doi:10.3906/bot-1310-6
20. Gulcin, I. (2009). Antioxidant activity of L-adrenaline: a structure-activity insight. chemicobiological interactions, 179(2), 71-80.
21. Gulcin, İ. (2020). Antioxidants and antioxidant methods: an updated overview. Archives of Toxicology, 94(3), 651-715. doi:10.1007/s00204-020-02689-3
22. Gulçin, İ., Taslimi, P., Aygün, A., Sadeghian, N., Bastem, E., Kufrevioglu, O. I., . . . Şen, F. (2018). Antidiabetic and antiparasitic potentials: Inhibition effects of some natural antioxidant compounds on α-glycosidase, α-amylase and human glutathione S-transferase enzymes. International Journal of Biological Macromolecules, 119, 741-746. https://doi.org/10.1016/j.ijbiomac.2018.08.001
23. Guven, L., Ozgen, U., Secen, H., Sener, S. Ö., Badem, M., Celik, G., & Yayli, N. (2021). Phytochemical studies on the seeds, pseudofruits, and roots of Rosa pimpinellifolia. Journal of Research in Pharmacy, 25(2), 153-163.
24. Gülçin, I. (2008). Measurement of antioxidant ability of melatonin and serotonin by the DMPD and CUPRAC methods as trolox equivalent. Journal of Enzyme Inhibition and Medicinal Chemistry, 23(6), 871-876. https://doi.org/10.1080/14756360701626223
25. Gülçin, İ., Elias, R., Gepdiremen, A., Taoubi, K., & Köksal, E. (2009). Antioxidant secoiridoits from fringe tree (Chionanthus virginicus L.). Wood science and technology, 43(3), 195-212.
26. Gülçin, İ., Gören, A. C., Taslimi, P., Alwasel, S. H., Kılıc, O., & Bursal, E. (2020). Anticholinergic, antidiabetic and antioxidant activities of Anatolian pennyroyal (Mentha pulegium)-analysis of its polyphenol contents by LC-MS/MS. Biocatalysis and Agricultural Biotechnology, 23, 101441. https://doi.org/10.1016/j.bcab.2019.101441
27. Gülçin, İ., Huyut, Z., Elmastaş, M., & Aboul-Enein, H. Y. (2010). Radical scavenging and antioxidant activity of tannic acid. Arabian journal of chemistry, 3(1), 43-53.
28. Güven, L., Erturk, A., Miloğlu, F. D., Alwasel, S., & Gulcin, İ. (2023). Screening of Antiglaucoma, Antidiabetic, Anti-Alzheimer, and Antioxidant Activities of Astragalus alopecurus Pall—Analysis of Phenolics Profiles by LC-MS/MS. Pharmaceuticals, 16(5), 659.
29. Izzotti, A., Bagnis, A., & Saccà, S. C. (2006). The role of oxidative stress in glaucoma. Mutation Research/Reviews in Mutation Research, 612(2), 105-114. https://doi.org/10.1016/j.mrrev.2005.11.001
30. Karagecili, H., Yılmaz, M. A., Ertürk, A., Kiziltas, H., Güven, L., Alwasel, S. H., & Gulcin, İ. (2023). Comprehensive Metabolite Profiling of Berdav Propolis Using LC-MS/MS: Determination of Antioxidant, Anticholinergic, Antiglaucoma, and Antidiabetic Effects. Molecules, 28(4), 1739.
31. Kasapligil, G., Nalbantsoy, A., & Medine, E. I. (2020). Davisianoside B ve aristatoside C monodezmozidik saponinlerin akciğer kanser hücreleri üzerinde sitotoksik ve apoptotik aktivitesinin incelenmesi. (yuksek lisans tezi), EGE UNIVERSITESI, IZMIR. Kırmızıgül, S., Sarıkahya, N. B., Sümbül, H., Göktürk, R. S., Yavaşoğlu, N. Ü., Pekmez, M., & Arda, N. (2012). Fatty acid profile and biological data of four endemic Cephalaria species grown in Turkey. Records of Natural Products, 6(2), 151-155.
32. Kiziltas, H., Bingol, Z., Goren, A. C., Pinar, S. M., Alwasel, S. H., & Gulcin, I. (2021). LC-HRMS profiling of phytochemicals, antidiabetic, anticholinergic and antioxidant activities of evaporated ethanol extract of Astragalus brachycalyx Fischer. Journal of Chemical Metrology, 15(2), 135-151. https://doi.org/10.25135/jcm.62.2107.2155
33. Kiziltas, H., Bingol, Z., Goren, A. C., Pinar, S. M., Ortaakarsu, A. B., Alwasel, S. H., & Gulcin, İ. (2022). Comprehensive metabolic profiling of Acantholimon caryophyllaceum using LC–HRMS and evaluation of antioxidant activities, enzyme inhibition properties and molecular docking studies. South African Journal of Botany, 151, 743-755. https://doi.org/10.1016/j.sajb.2022.10.048
34. Mbhele, N., Balogun, F. O., Kazeem, M. I., & Ashafa, T. (2015). In vitro studies on the antimicrobial, antioxidant and antidiabetic potential of Cephalaria gigantea. ||| Bangladesh Journal of Pharmacology, 10(1), 214-221.
35. Okkay, U., Ferah Okkay, I., Cicek, B., Aydin, I. C., Ertugrul, M. S., Bayram, C., . . . Hacimuftuoglu, A. (2021). Achillea millefolium alleviates testicular damage in paclitaxel-intoxicated rats via attenuation of testicular oxido-inflammatory stress and apoptotic responses. Andrologia, 53(5), e14028. doi:10.1111/and.14028
36. Oyaizu, M. (1986). Studies on Products of Browning Reaction Antioxidative Activities of Products of Browning Reaction Prepared from Glucosamine. The Japanese Journal of Nutrition and Dietetics, 44(6), 307-315. https://doi.org/10.5264/eiyogakuzashi.44.307
37. Rahimi, A., Siavash Moghaddam, S., Ghiyasi, M., Heydarzadeh, S., Ghazizadeh, K., & Popović-Djordjević, J. (2019). The Influence of Chemical, Organic and Biological Fertilizers on Agrobiological and Antioxidant Properties of Syrian Cephalaria (Cephalaria Syriaca L.). Agriculture, 9(6), 122.
38. Re, R., Pellegrini, N., Proteggente, A., Pannala, A., Yang, M., & Rice-Evans, C. (1999). Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free radical biology and medicine, 26(9), 1231-1237. https://doi.org/10.1016/S0891-5849(98)00315-3
39. Rochette, L., Zeller, M., Cottin, Y., & Vergely, C. (2014). Diabetes, oxidative stress and therapeutic strategies. Biochimica et Biophysica Acta (BBA) - General Subjects, 1840(9), 2709-2729. https://doi.org/10.1016/j.bbagen.2014.05.017
40. Saleem, H., Sarfraz, M., Ahsan, H. M., Khurshid, U., Kazmi, S. A. J., Zengin, G., . . . Ahemad, N. (2020). Secondary Metabolites Profiling, Biological Activities and Computational Studies of Abutilon figarianum Webb (Malvaceae). Processes, 8(3), 336.
41. Sarikahya, N. B. (2014). Aristatosides A-C, hederagenin-type triterpene saponins from Cephalaria aristata. Phytochem. Lett., 8, 149-155. doi:10.1016/j.phytol.2014.03.001
42. Sarikahya, N. B., Goren, A. C., & Kirmizigul, S. (2019). Simultaneous determination of several flavonoits and phenolic compounds in nineteen different Cephalaria species by HPLC-MS/MS. Journal of pharmaceutical and biomedical analysis, 173, 120-125.
43. Sarikahya, N. B., & Kirmizigul, S. (2010). Antimicrobial triterpenoid glycosides from Cephalaria scoparia. J. Nat. Prod., 73(5), 825-830. doi:10.1021/np900724u
44. SARIKAHYA, N. B., & Kirmizigul, S. (2010). Isolation and structural determination of Cephalaria gazipashensis, Cephalaria elmaliensis and Cephalaria scoparia glycosides and Their Biological Properties. (Doctor of philosophy), Ege University, Bornova-IZMIR.
45. Sarikahya, N. B., Nalbantsoy, A., Top, H., Gokturk, R. S., Sumbul, H., & Kirmizigul, S. (2018). Immunomodulatory, hemolytic and cytotoxic activity potentials of triterpenoid saponins from eight Cephalaria species. Phytomedicine, 38, 135-144. doi:10.1016/j.phymed.2017.11.009
46. Sarikahya, N. B., Ucar, E. O., Kayce, P., Gokturk, R. S., Sumbul, H., Arda, N., & Kirmizigul, S. (2015). Fatty acid composition and antioxidant potential of ten Cephalaria species. Records of Natural Products, 9(1), 116-123.
47. Sezer Senol, F., Orhan, I. E., Ozgen, U., Renda, G., Bulut, G., Guven, L., . . . Sekeroglu, N. (2016). Memory-vitalizing effect of twenty-five medicinal and edible plants and their isolated compounds. South African Journal of Botany, 102, 102-109. https://doi.org/10.1016/j.sajb.2015.07.011
48. Singleton, V. L., Orthofer, R., & Lamuela-Raventós, R. M. (1999). [14] Analysis of total phenols and other oxidation substrates and antioxidants by means of folin-ciocalteu reagent Methods in Enzymology (Vol. 299, pp. 152-178): Academic Press.
49. Sumer, G., & Kirmizigul, S. (2018). Phytochemical investigations of biologically active natural products from Cephalaria anatolica Schchian. (MASTER OF SCIENCE THESIS), Ege University, İzmir.
50. Topal, M., & Gulcin, I. (2014). Rosmarinic acid: a potent carbonic anhydrase isoenzymes inhibitor. Turkish Journal of Chemistry, 38(5), 894-902. doi:10.3906/kim-1403-5
51. Topal, M., & Gulcin, İ. (2022). Evaluation of the in vitro antioxidant, antidiabetic and anticholinergic properties of rosmarinic acid from rosemary (Rosmarinus officinalis L.). Biocatalysis and Agricultural Biotechnology, 43, 102417. https://doi.org/10.1016/j.bcab.2022.102417
52. Zengin, G., Atasagun, B., Zakariyyah Aumeeruddy, M., Saleem, H., Mollica, A., Babak Bahadori, M., & Mahomoodally, M. F. (2019). Phenolic profiling and in vitro biological properties of two Lamiaceae species (Salvia modesta and Thymus argaeus): A comprehensive evaluation. Industrial Crops and Products, 128, 308-314. https://doi.org/10.1016/j.indcrop.2018.11.027