Carduus nutans L.’ın LC-MS/MS ile Fitokimyasal Analizi, Antioksidan, Antidiyabetik, Antiglokom ve Anti-Alzheimer Etkileri

Abstract

Carduus nutans L. (Asteraceae), halk arasında “Eşek dikeni” olarak bilinir ve kökleri cüzzam ve kabızlık tedavisinde, yaprakları ise sıtma tedavisinde kullanılmaktadır. Yapılan çalışmada Carduus nutans metanol ekstraktı (MECN) antioksidan aktivitesi ve enzim inhibisyon etkileri spektrofotometrik olarak belirlenmiştir. Antioksidan deneylerinden üçü Fe3+, Cu2+ ve Fe3+-TPTZ indirgeme deneyi olup sırasıyla 20 μg/mL’de ki absorbansları λ700: 0,470, λ450: 0,521 ve λ593: 0,671’dir. Diğer antioksidan deneyleri ise DPPH, ABTS ve DMPD radikalleri süpürme deneyi olup IC50 değerleri sırasıyla 115,52 μg/mL, 37,71 μg/mL, 85,47 μg/mL’dir. MECN’nin asetilkolinesteraz (AChE), α-glikozidaz (α-gli) ve karbonik anhidraz I-II (KA I-II) enzimlerine karşı inhibisyon etkilerinin IC50 değerleri sırasıyla 3,28 μg/mL, 8,37 μg/mL, 18,74 μg/mL ve 21,83 μg/mL’dir. MECN’nin toplam fenolik madde miktarı 153,83 μg/g GAE, toplam flavonoid madde miktarı ise 221,0 μg/g KE’dir. Ayrıca MECN’nin LC-MS/MS analizi yapılmış 7,07 µg/g fumarik asit, 2,72 µg/g kinik asit, 1,09 µg/g klorojenik asit majör madde olarak tespit edilmiştir. C. nutans metanol ekstraktı yüksek antioksidan aktiviteye sahiptir ve AChE, α-gli ve KA I-II enzimlerine karşı iyi bir inhibisyon etkisine sahiptir.

Keywords

• Carduus nutans
• Antioksidan aktivite
• Enzim inhibisyonu
• LC-MS/MS analizi

Thanks

Çalışmanın yapılmasında danışmanlık yapan ve laboratuvar imkanlarının kullanılmasına imkan sağlayan Prof. Dr. İlhami Gülçin’e teşekkürlerimizi sunarız.

Phytochemical Analysis of Carduus nutans L. via LC-MS/MS, Antioxidant, Antidiabetic, Antiglaucoma and Anti-Alzheimer's Effects

Abstract

Carduus nutans L. (Asteraceae) is popularly known as "Eşek dikeni" and its roots are used in the treatment of leprosy and constipation, and its leaves are used in the treatment of malaria. In the study, antioxidant activity and enzyme inhibition effects of Carduus nutans methanol extract (MECN) were determined spectrophotometrically. Three of the antioxidant assays are Fe3+, Cu2+ and Fe3+-TPTZ reduction experiments, and their absorbances at 20 μg/mL are λ700:0,470, λ450: 0,521, and λ593: 0,671, respectively. Other antioxidant assays are DPPH, ABTS and DMPD radical scavenging experiments and IC50 values are 115.52 μg/mL, 37.71 μg/mL, and 85.47 μg/mL, respectively. The IC50 values of the inhibitory effects of MECN against acetylcholinesterase (AChE), α-glucosidase (α-gly) and carbonic anhydrase I-II (CA I-II) are 3.28 μg/mL, 8.37 μg/mL, 18.74 μg/mL, and 21.83 μg/mL, respectively. The total phenolic substance amount of MECN is 153.83 μg/g GAE, and total flavonoid substance amount is 221.0 μg/g QE. Additionally, LC-MS/MS analysis of MECN was performed and 7.07 µg/g fumaric acid, 2.72 µg/g quinic acid and 1.09 µg/g chlorogenic acid were determined as majör components. C. nutans methanol extract has high antioxidant activity and a good inhibition effect against AChE, α-gly, and CA I- II enzymes.

Keywords

• Carduus nutans
• Antioxidant activity
• Enzym inhibition
• LC-MS/MS analysis

References

1. Adali A., Yirün A., Koçer Gümüşel B., Erkekoğlu Ü. The possible effects of biological agents on the development of Alzheimer’s disease. Journal of Faculty of Pharmacy of Ankara University 2020; 44(1): 167-187.
2. Ahamad J., Naquvi K.J., Mir SR., Ali M., Shuaib M. Review on role of natural αlpha-glucosidase inhibitors for management of diabetes mellitus. International Journal of Biomedical Research 2011; (2): 374-380.
3. Akao M., Kuroda K. Antifungal activity of fumaric acid in mice infected with Candida albicans. Chemical and pharmaceutical bulletin 1991; 39(11): 3077-3078.
4. Al-Shammari LA., Hassan WHB., Al-Youssef HM. Phytochemical and biological studies of Carduus pycnocephalus L. Journal of Saudi Chemical Society 2015; 19(4): 410-416.
5. Atıcı Y., Bulduk İ. Optimization the ultrasonic assisted extraction of chlorogenic acid from the artichoke (Cynara Scolymus) leaf with response surface methodology and total phenolic contents. Hacettepe Journal of Biology and Chemistry 2020; 48(2): 147-157.
6. Blois MS. Antioxidant determinations by the use of a stable free radical. Nature 1958; 181(4617): 1199-1200.
7. Bursal E., Aras A., Kılıç Ö., Taslimi P., Gören AC., Gülçin İ. Phytochemical content, antioxidant activity, and enzyme inhibition effect of Salvia eriophora Boiss. & Kotschy against acetylcholinesterase, α-amylase, butyrylcholinesterase, and α-glycosidase enzymes. Journal of Food Biochemistry 2019; 43(3): e12776.
8. Çakmakçı S., Gülçin İ., Gündoğdu E., Ertem Öztekin H., Taslimi P. The Comparison with Commercial Antioxidants, Effects on Colour, and Sensory Properties of Green Tea Powder in Butter. Antioxidants 2023; 12(8): 1522.

9. Davis P.H. Flora of Turkey and the East Aegean Islands. Edinburgh University Press 1970; 3.
10. Deliorman Orhan D., Yeå Ilada E., Ergun F., Tsuchiya K., Takaishi Y. The antioxidant activity of the extracts and fractions from Carduus nutans L. Gazi Universitesi Eczacilik Fakultesi Dergisi 2002; 19: 61-66.
11. Demir Z., Türkan F. Asetilkolinesteraz ve Bütirilkolinesteraz Enzimlerinin Alzheimer Hastalığı ile İlişkisi. Journal of the Institute of Science and Technology 2022; 12(4): 2386-2395.
12. Dimitrova-Dyulgerova I., Zhelev I., Mihaylova D. Phenolic Profile and In vitro Antioxidant Activity of Endemic Bulgarian Carduus Species. Pharmacognosy Magazine 2015; 11(4): S575-579.
13. Durmaz L., Kiziltas H., Guven L., Karagecili H., Alwasel S., Gulcin İ. Antioxidant, antidiabetic, anticholinergic, and antiglaucoma effects of magnofluorine. Molecules 2022; 27(18): 5902.
14. Ellman GL., Courtney KD., Andres V., Featherstone R.M. A new and rapid colorimetric determination of acetylcholinesterase activity. Biochemical Pharmacology 1961; 7(2): 88-95.
15. Fogliano V., Verde V., Randazzo G., Ritieni A. Method for measuring antioxidant activity and its application to monitoring the antioxidant capacity of wines. Journal of Agricultural and Food Chemistry 1999; 47(3): 1035-1040.
16. Göcer H., Akıncıoğlu A., Göksu S., Gülçin İ. Carbonic anhydrase inhibitory properties of phenolic sulfonamides derived from dopamine related compounds. Arabian Journal Of Chemistry 2017; 10(3): 398-402.
17. Gulcin I., Kaya R., Goren A.C., Akincioglu H., Topal M., Bingol Z., Cetin Çakmak K., Ozturk Sarikaya SB., Durmaz L., Alwasel S. Anticholinergic, antidiabetic and antioxidant activities of cinnamon (Cinnamomum verum) bark extracts: polyphenol contents analysis by LC-MS/MS. International Journal of Food Properties 2019; 22(1): 1511-1526.
18. Gulcin İ. Antioxidants and antioxidant methods: An updated overview. Archives of Toxicology 2020; 94(3): 651-715.
19. Gulcin İ., Alwasel S.H. DPPH radical scavenging assay. Processes 2023; 11(8): 2248.
20. Gulçin I. Measurement of antioxidant ability of melatonin and serotonin by the DMPD and CUPRAC methods as trolox equivalent. Journal Enzyme Inhibition and Medicinal Chemistry 2008; 23(6): 871-876.
21. Güven L., Erturk A., Koca M., Gulcin I. Phenolic compounds of Phlomis tuberosa by LC–MS/MS‐determination of antioxidant activity, molecular docking, and enzyme ınhibition profiles. ChemistrySelect 2023a; 8(48): e202303101.
22. Güven L., Erturk A., Miloğlu FD., Alwasel S., Gulcin İ. Screening of antiglaucoma, antidiabetic, anti-alzheimer, and antioxidant activities of Astragalus alopecurus Pall—analysis of phenolics profiles by LC-MS/MS. Pharmaceuticals 2023b; 16(5): 659.
23. Güven L., Ertürk A., Yılmaz MA., Alwasel S., Gülçin İ. Alchemilla pseudocartalinica Juz: phytochemical screening by UPLC-MS/MS, molecular docking, anti-oxidant, anti-diabetic, anti-glaucoma, and anti-Alzheimer effects. Records Of Natural Products 2024; 18(2): 251-272
24. Inci H., Izol E., Yilmaz MA., Ilkaya M., Bingöl Z., Gülçin I. Comprehensive phytochemical content by LC/MS/MS and anticholinergic, antiglaucoma, antiepilepsy, and antioxidant activity of apilarnil (Drone Larvae). Chemistry & Biodiversity 2023; 20(10): e202300654.
25. Kadereit J.W., Jeffrey C. The families and genera of vascular plants, Springer 2007.
26. Kaplan ABU., Cetin M., Bayram C., Yildirim S., Taghizadehghalehjoughi A., Hacimuftuoglu A. In vivo evaluation of nanoemulsion formulations for metformin and repaglinide alone and combination. Journal of Pharmaceutical Sciences 2023; 112(5): 1411-1426.
27. Karagecili H., İzol E., Kirecci E., Gulcin İ. Determination of antioxidant, anti-alzheimer, antidiabetic, antiglaucoma and antimicrobial effects of zivzik pomegranate (Punica granatum) -; A chemical profiling by LC-MS/MS. Life 2023a; 13(3): 735.
28. Karagecili H., Yılmaz MA., Ertürk A., Kiziltas H., Güven L., Alwasel SH., Gulcin İ. Comprehensive metabolite profiling of Berdav propolis using LC-MS/MS: determination of antioxidant, anticholinergic, antiglaucoma, and antidiabetic effects. Molecules 2023b; 28(4): 1739.
29. Keleş E., Özalevli S. Alzheimer’s disease and treatment approaches. İzmir Kâtip Çelebi Üniversitesi Sağlık Bilimleri Fakültesi Dergisi 2018; 3(2): 39-42.
30. Kiziltas H., Bingol Z., Goren AC., Pinar S.M., Ortaakarsu AB., Alwasel SH., Gulcin İ. 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 2022a; 151: 743-755.
31. Kiziltas H., Goren A.C., Alwasel SH., Gulcin İ. Sahlep (Dactylorhiza osmanica): Phytochemical analyses by LC-HRMS, molecular docking, antioxidant activity, and enzyme ınhibition profiles. Molecules 2022b; 27(20): 6907.
32. Koca M., Gülçin İ., Üç EM., Bilginer S., Aydın AS. Evaluation of antioxidant potentials and acetylcholinesterase inhibitory effects of some new salicylic acid-salicylamide hybrids. Journal of the Iranian Chemical Society 2023; 20(7): 1535-1543.
33. Kocaoğlu E. Kinik asidin sitotoksik/genotoksik ve antigenotoksik etkilerinin sağlıklı akciğer epitel hücre hatlarında belirlenmesi, Uludag Üniversitesi Fen Bilimleri Enstitüsü Yüksek Lisans Tezi sayfa no:8, Bursa, Türkiye, 2014.
34. Kozyra M., Komsta Ł., Wojtanowski K. Analysis of phenolic compounds and antioxidant activity of methanolic extracts from inflorescences of Carduus sp. Phytochemistry Letters 2019; 31: 256-262.
35. Kozyra M., Kukula‐Koch W., Szymański M. Phenolic composition of ınflorescences of Carduus nutans L. Chemistry & Biodiversity 2022; 19(3): e202100827.
36. Kucukoglu K., Gul H.I., Taslimi P., Gulcin I., Supuran CT. Investigation of inhibitory properties of some hydrazone compounds on hCA I, hCA II and AChE enzymes. Bioorganic Chemistry 2019; 86: 316-321.
37. Li X., Feng X., Sun X., Hou N., Han F., Liu Y. Global, regional, and national burden of Alzheimer's disease and other dementias, 1990–2019. Frontiers in Aging Neuroscience 2022; 14.
38. Miloglu F.D., Akpınar A., Güven L., Demirkaya A.K., Gundogdu G., Nalcı KA., Hacımuftuoglu A. Evaluation the effects of Helichrysum plicatum subsp. pseudoplicatum on an in-vitro wound model using human dermal fibroblast cells. International Journal of Low Extremity Wounds 2023; 22(2): 401-408.
39. Mohammed F., Sibley P., Abdulwali N., Guillaume D. Nutritional, pharmacological, and sensory properties of maple syrup: A comprehensive review. Heliyon 2023.
40. Mrowietz U., Morrison P.J., Suhrkamp I., Kumanova M., Clement B. The pharmacokinetics of fumaric acid esters reveal their in vivo effects. Trends in Pharmacological Sciences 2018; 39(1): 1-12.
41. Oyaizu M. Studies on products of browning reaction antioxidative activities of products of browning reaction prepared from glucosamine. The Japanese Journal of Nutrition and Dietetics 1986; 44(6): 307-315.
42. Özcan K. Determination of biological activity of Carduus lanuginosus: an endemic plant in Turkey. International Journal of Environmental Health Research 2021; 31(1): 45-53.
43. Öztürk G.B., Karan M.A. Physiopathology of Alzheimer's disease. Klinik gelişim 2009; 22(3): 36-45.
44. Re R., Pellegrini N., Proteggente A., Pannala A., Yang M., Rice-Evans C. Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radical Biology and Medicine 1999; 26(9): 1231-1237.
45. Riyaphan J., Pham D.-C., Leong M.K., Weng C.-F. In silico approaches to ıdentify polyphenol compounds as α-glucosidase and α-amylase inhibitors against type-II Diabetes. Biomolecules 2021; 11(12): 1877.
46. Scozzafava A., Supuran CT. Glaucoma and the applications of carbonic anhydrase ınhibitors. In: Frost S., McKenna R. (eds). Carbonic anhydrase: mechanism, regulation, links to disease, and industrial applications. Subcellular Biochemistry, Dordrecht: Springer 2014; 75, 349-359.
47. Singleton VL., Orthofer R., Lamuela-Raventós RM. Analysis of total phenols and other oxidation substrates and antioxidants by means of Folin-Ciocalteu reagent. In: Packer L. (ed.). Oxidants and antioxidants, part A. Methods in Enzymology, New York: Academic Press 1999;. 299,152-178.
48. Supuran CT. Carbonic Anhydrases and Metabolism. Metabolites 2018; 8(2): 25.
49. Şenol H., Çelik Turgut G., Şen A., Sağlamtaş R., Tuncay S., Gülçin İ., Topçu G. Synthesis of nitrogen-containing oleanolic acid derivatives as carbonic anhydrase and acetylcholinesterase inhibitors. Medicinal Chemistry Research 2023; 32(4): 694-704.
50. Tao Y., Zhang Y., Cheng Y., Wang Y. Rapid screening and identification of α-glucosidase inhibitors from mulberry leaves using enzyme-immobilized magnetic beads coupled with HPLC/MS and NMR. Biomedical Chromatography 2013; 27(2): 148-155.
51. Taslimi P., Gulcin I., Ozgeris B., Goksu S., Tumer F., Alwasel SH., Supuran CT. The human carbonic anhydrase isoenzymes I and II (hCA I and II) inhibition effects of trimethoxyindane derivatives. Journal of Enzyme Inhibion Medicinal Chemistry 2016; 31(1): 152-157.
52. Tübives. (2024). "Carduus sp." Retrieved 16.05.2024, 2024, from http://194.27.225.161/yasin/tubives/index.php?sayfa=karsilastir.
53. Yilmaz MA. Simultaneous quantitative screening of 53 phytochemicals in 33 species of medicinal and aromatic plants: A detailed, robust and comprehensive LC–MS/MS method validation. Industrial Crops and Products 2020; 149: 112347.
54. Zheleva-Dimitrova D., Zhelev I., Dimitrova-Dyulgerova I. Antioxidant activity of some Carduus species growing in Bulgaria. Free Radicals and Antioxidants 2011; 1(4): 15-20.