Metabolomic Approach for Rapid Identification of Antioxidants in Clinacanthus nutans Leaves with Liver Protective Potential
Abstract
:1. Introduction
2. Results
2.1. Determination of Phytochemical Contents
2.2. Antioxidant Potentials of Clinacanthus nutans Leaf and Stem Methanolic Extracts
2.3. Cytotoxic Effect of Clinacanthus nutans Leaf and Stem Methanolic Extracts on HepG2 and HaCaT Cell Lines
2.4. Direct and Protective Effects of Clinacanthus nutans Extracts on Intracellular Reactive Oxygen Species (ROS) Generation of HepG2 Cell Line
2.5. 1H-NMR Spectra of Plant Extracts and Metabolite Identification
2.6. Relationship between Bioactivities and Plant Metabolites Using Partial Least-Square Analysis (PLS)
3. Discussion
4. Materials and Methods
4.1. Chemicals and Reagents
4.2. Preparation of Clinacanthus nutans Leaf (CNL) and Stem Extracts (CNS)
4.3. Determination of Phytochemical Contents
4.3.1. Total Phenolic Content
4.3.2. Total Flavonoid Content
4.4. Chemical-Based Antioxidant Assays
4.4.1. DPPH Radical Scavenging Activity Assay
4.4.2. ABTS Radical Scavenging Activity Assay
4.4.3. Ferric Ion Chelating Activity Assay
4.4.4. Ferric Reducing Antioxidant Power (FRAP) Assay
4.5. Cell-Based Antioxidant Assays
4.5.1. Cytoxicity Assays
4.5.2. ROS Direct
4.5.3. ROS Protection
4.6. 1H NMR-Based Metabolomic Study
4.7. Statistical Analysis
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Sample Availability
References
- Nakano, M.; Yatsuhashi, H.; Bekki, S.; Takami, Y.; Tanaka, Y.; Yoshimaru, Y.; Honda, K.; Komorizono, Y.; Harada, M.; Shibata, M.; et al. Trends in hepatocellular carcinoma incident cases in Japan between 1996 and 2019. Sci. Rep. 2022, 12, 1517. [Google Scholar] [CrossRef] [PubMed]
- Musolino, V.; Gliozzi, M.; Bombardelli, E.; Nucera, S.; Carresi, C.; Maiuolo, J.; Mollace, R.; Paone, S.; Bosco, F.; Scarano, F.; et al. The synergistic effect of Citrus bergamia and Cynara cardunculus extracts on vascular inflammation and oxidative stress in non-alcoholic fatty liver disease. J. Tradit. Complement. Med. 2020, 10, 268–274. [Google Scholar] [CrossRef]
- Takaki, A.; Kawai, D.; Yamamoto, K. Multiple hits, including oxidative stress, as pathogenesis and treatment target in non-alcoholic steatohepatitis (NASH). Int. J. Mol. Sci. 2013, 14, 20704–20728. [Google Scholar] [CrossRef] [Green Version]
- Masarone, M.; Rosato, V.; Dallio, M.; Gravina, A.; Aglitti, A.; Loguercio, C.; Federico, A.; Persico, M. Role of oxidative stress in pathophysiology of nonalcoholic fatty liver disease. Oxidative Med. Cell. Longev. 2018, 2018, 9547613. [Google Scholar] [CrossRef] [PubMed]
- Bagherniya, M.; Nobili, V.; Blesso, C.; Sahebkar, A. Medicinal plants and bioactive natural compounds in the treatment of non-alcoholic fatty liver disease: A clinical review. Pharmacol. Res. 2018, 130, 213–240. [Google Scholar] [CrossRef] [PubMed]
- Abu-Serie, M.; Habashy, N. Vitis vinifera polyphenols from seedless black fruit act synergistically to suppress hepatotoxicity by targeting necroptosis and pro-fibrotic mediators. Sci. Rep. 2020, 10, 2452. [Google Scholar] [CrossRef] [Green Version]
- Lin, C.; Chen, H.; Lung, C.; Chen, H. Recent advancement in anticancer activity of Clinacanthus nutans (Burm. f.) Lindau. Evid.-Based Complement. Altern. Med. 2021, 2021, 1–13. [Google Scholar] [CrossRef]
- Azemi, A.; Mokhtar, S.; Rasool, A. Clinacanthus nutans leaves extract reverts endothelial dysfunction in type 2 diabetes rats by improving protein expression of eNOS. Oxidative Med. Cell. Longev. 2020, 2020, 7572892. [Google Scholar] [CrossRef] [PubMed]
- Ahmad Azam, A.; Ismail, I.; Kumari, Y.; Shaikh, M.; Abas, F.; Shaari, K. The anti-neuroinflammatory effects of Clinacanthus nutans leaf extract on metabolism elucidated through 1H NMR in correlation with cytokines microarray. PLoS ONE 2020, 15, e0238503. [Google Scholar] [CrossRef]
- Nik Abd Rahman, N.; Nurliyana, M.; Afiqah, M.; Osman, M.; Hamid, M.; Lila, M. Antitumor and antioxidant effects of Clinacanthus nutans Lindau in 4 T1 tumor-bearing mice. BMC Complement. Altern. Med. 2019, 19, 340. [Google Scholar] [CrossRef]
- Kong, H.S.; Abdullah Sani, N. Antimicrobial properties of the acetone leaves and stems extracts of Clinacanthus nutans from three different samples/areas against pathogenic microorganisms. Int. Food Res. J. 2018, 25, 1698–1702. [Google Scholar]
- Khoo, L.; Audrey Kow, S.; Lee, M.; Tan, C.; Shaari, K.; Tham, C.; Abas, F. A comprehensive review on phytochemistry and pharmacological activities of Clinacanthus nutans (Burm.f.) Lindau. Evid. Based Complement. Altern. Med. 2018, 2018, 1–39. [Google Scholar] [CrossRef] [Green Version]
- Yeo, B.; Yap, Y.; Koh, R.; Ng, K.; Chye, S. Medicinal properties of Clinacanthus nutans: A review. Trop. J. Pharm. Res. 2018, 17, 375. [Google Scholar] [CrossRef] [Green Version]
- Hong, J.; Yang, L.; Zhang, D.; Shi, J. Plant Metabolomics: An indispensable system biology tool for plant science. Int. J. Mol. Sci. 2016, 17, 767. [Google Scholar] [CrossRef] [PubMed]
- Emwas, A.; Roy, R.; McKay, R.; Tenori, L.; Saccenti, E.; Gowda, G.; Raftery, D.; Alahmari, F.; Jaremko, L.; Jaremko, M.; et al. NMR spectroscopy for metabolomics research. Metabolites 2019, 9, 123. [Google Scholar] [CrossRef] [Green Version]
- Khoo, L.; Kow, A.; Maulidiani, M.; Ang, M.; Chew, W.; Lee, M.; Tan, C.P.; Shaari, K.; Tham, C.L.; Abas, F. 1H-NMR metabolomics for evaluating the protective effect of Clinacanthus nutans (Burm. f) Lindau water extract against nitric oxide production in LPS-IFN-γactivated RAW 264.7 macrophages. Phytochem. Anal. 2018, 30, 46–61. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Khoo, L.; Mediani, A.; Zolkeflee, N.; Leong, S.; Ismail, I.; Khatib, A.; Shaari, K.; Abas, F. Phytochemical diversity of Clinacanthus nutans extracts and their bioactivity correlations elucidated by NMR based metabolomics. Phytochem. Lett. 2015, 14, 123–133. [Google Scholar] [CrossRef]
- Albuquerque, B.; Heleno, S.; Oliveira, M.; Barros, L.; Ferreira, I. Phenolic compounds: Current industrial applications, limitations and future challenges. Food Funct. 2021, 12, 14–29. [Google Scholar] [CrossRef] [PubMed]
- Kaurinovic, B.; Vastag, D. Flavonoids and phenolic acids as potential natural antioxidants. In Antioxidants; InTech: London, UK, 2019. [Google Scholar] [CrossRef] [Green Version]
- Minatel, I.; Borges, C.; Ferreira, M.; Gomez, H.; Chen, C.; Lima, G. Phenolic Compounds: Functional properties, impact of processing and bioavailability. Phenolic Compd. Biol. Act. 2017, 8, 1–24. [Google Scholar] [CrossRef] [Green Version]
- Tan, L.; Khaw, K.; Ong, Y.; Khan, T.; Lee, L.; Lee, W.; Goh, B. An overview of Clinacanthus nutans (Burm. f.) Lindau as a medicinal plant with diverse pharmacological values. In Plant-Derived Bioactives; Spinger: Berlin/Heidelberg, Germany, 2020; pp. 461–491. [Google Scholar] [CrossRef]
- Mustapa, A.; Martin, Á.; Mato, R.; Cocero, M. Extraction of phytocompounds from the medicinal plant Clinacanthus nutans Lindau by microwave-assisted extraction and supercritical carbon dioxide extraction. Ind. Crops Prod. 2015, 74, 83–94. [Google Scholar] [CrossRef]
- Sarega, N.; Imam, M.; Ooi, D.; Chan, K.; Md Esa, N.; Zawawi, N.; Ismail, M. Phenolic rich extract from Clinacanthus nutans attenuates hyperlipidemia-associated oxidative stress in rats. Oxidative Med. Cell. Longev. 2016, 2016, 4137908. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Hamid, H.; Yahya, I.; Yusoff, M.; Zareen, S. Bioassay-guided isolation and antioxidant activity of sulfur-containing compounds from Clinacanthus nutans. J. Chin. Chem. Soc. 2016, 63, 1033–1037. [Google Scholar] [CrossRef] [Green Version]
- Zhang, Q.; Lin, L.; Ye, W. Techniques for extraction and isolation of natural products: A comprehensive review. Chin. Med. 2018, 13, 1–26. [Google Scholar] [CrossRef] [Green Version]
- Yang, H.S.; Peng, T.W.; Madhavan, P.; Abdul Shukkoor, M.; Akowuah, G. Phytochemical analysis and antibacterial activity of methanolic extract of Clinacanthus nutans leaf. Int. J. Drug Dev. Res. 2013, 5, 349–355. [Google Scholar]
- Yusof, Z.; Ramasamy, S.; Mahmood, N.; Yaacob, J. Vermicompost supplementation improves the stability of bioactive anthocyanin and phenolic compounds in Clinacanthus nutans Lindau. Molecules 2018, 23, 1345. [Google Scholar] [CrossRef] [Green Version]
- Ahmadinejad, F.; Geir Møller, S.; Hashemzadeh-Chaleshtori, M.; Bidkhori, G.; Jami, M. Molecular mechanisms behind free radical scavengers’ function against oxidative stress. Antioxidants 2017, 6, 51. [Google Scholar] [CrossRef] [PubMed]
- Engwa, G. Free radicals and the role of plant phytochemicals as antioxidants against oxidative stress-related diseases. Phytochem. Source Antioxid. Role Dis. Prev. 2018, 7, 49–74. [Google Scholar] [CrossRef] [Green Version]
- Haida, Z.; Nakasha, J.; Hakiman, M. In vitro responses of plant growth factors on growth, yield, phenolics content and antioxidant activities of Clinacanthus nutans (Sabah Snake Grass). Plants 2020, 9, 1030. [Google Scholar] [CrossRef]
- Flora, S. Structural, chemical and biological aspects of antioxidants for strategies against metal and metalloid exposure. Oxidative Med. Cell. Longev. 2009, 2, 191–206. [Google Scholar] [CrossRef]
- Killian, B.; Yuan, T.; Tsai, C.; Chiu, T.; Chen, Y.; Chan, C. Emission-related heavy metal associated with oxidative stress in children: Effect of antioxidant intake. Int. J. Environ. Res. Public Health 2020, 17, 3920. [Google Scholar] [CrossRef]
- Vajrabhaya, L.; Korsuwannawong, S. Cytotoxicity evaluation of Clinacanthus nutans through dimethylthiazol diphenyltetrazolium bromide and neutral red uptake assays. Eur. J. Dent. 2016, 10, 134–138. [Google Scholar] [CrossRef] [PubMed]
- Mohd Roslan, S.; Zakaria, Y.; Abdullah, H. Cytotoxicity of Clinacanthus nutans and mechanism of action of its active fraction towards human cervical cancer cell line, HeLA. J. Sains Kesihat. Malays. 2018, 16, 39–50. [Google Scholar] [CrossRef]
- Liew, S.; Stanbridge, E.; Yusoff, K.; Shafee, N. Hypoxia affects cellular responses to plant extracts. J. Ethnopharmacol. 2012, 144, 453–456. [Google Scholar] [CrossRef]
- Kim, H.; Xue, X. Detection of total reactive oxygen species in adherent cells by 2′,7′-dichlorodihydrofluorescein diacetate staining. J. Vis. Exp. 2020, 160, e60682. [Google Scholar] [CrossRef]
- Zhang, J.; Wang, X.; Vikash, V.; Ye, Q.; Wu, D.; Liu, Y.; Dong, W. ROS and ROS-mediated cellular signaling. Oxidative Med. Cell. Longev. 2016, 2016, 4350965. [Google Scholar] [CrossRef] [Green Version]
- Kučera, O.; Endlicher, R.; Roušar, T.; Lotková, H.; Garnol, T.; Drahota, Z.; Červinková, Z. The effect of tert-butyl hydroperoxide-induced oxidative stress on lean and steatotic rat hepatocytes in vitro. Oxidative Med. Cell. Longev. 2014, 2014, 752506. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Domanski, A.; Lapshina, E.; Zavodnik, I. Oxidative processes induced by tert-butyl hydroperoxide in human red blood cells: Chemiluminescence studies. Biochemistry 2005, 70, 761–769. [Google Scholar] [CrossRef]
- Crane, D.; Häussinger, D.; Graf, P.; Sies, H. Decreased flux through pyruvate dehydrogenase by thiol oxidation during t-butyl hydroperoxide metabolism in perfused rat liver. Hoppe-Seyler’s Z. Für Physiol. Chem. 1983, 364, 977–988. [Google Scholar] [CrossRef] [PubMed]
- Radić, K.; Vinković Vrček, I.; Pavičić, I.; Čepo, D. Cellular antioxidant activity of Olive Pomace extracts: Impact of gastrointestinal digestion and cyclodextrin encapsulation. Molecules 2020, 25, 5027. [Google Scholar] [CrossRef]
- Kim, Y.; Hwang, J.; Sung, S.; Jeon, Y.; Jeong, J.; Jeon, B.; Moon, S.-H.; Park, P.-J. Antioxidant activity and protective effect of extract of Celosia cristata L. flower on tert-butyl hydroperoxide-induced oxidative hepatotoxicity. Food Chem. 2015, 168, 572–579. [Google Scholar] [CrossRef]
- Grauzdytė, D.; Pukalskas, A.; Viranaicken, W.; El Kalamouni, C.; Venskutonis, P. Protective effects of Phyllanthus phillyreifolius extracts against hydrogen peroxide induced oxidative stress in HEK293 cells. PLoS ONE 2018, 13, e0207672. [Google Scholar] [CrossRef] [Green Version]
- Martín, M.; Ramos, S.; Mateos, R.; Granado Serrano, A.; Izquierdo-Pulido, M.; Bravo, L.; Goya, L. Protection of human HepG2 cells against oxidative stress by cocoa phenolic extract. J. Agric. Food Chem. 2008, 56, 7765–7772. [Google Scholar] [CrossRef]
- Francenia Santos-Sánchez, N.; Salas-Coronado, R.; Villanueva-Cañongo, C.; Hernández-Carlos, B. Antioxidant compounds and their antioxidant mechanism. Antioxidants 2019, 10, 1–29. [Google Scholar] [CrossRef] [Green Version]
- Alam, A.; Ferdosh, S.; Ghafoor, K.; Hakim, A.; Juraimi, A.; Khatib, A.; Sarker, Z. Clinacanthus nutans: A review of the medicinal uses, pharmacology and phytochemistry. Asian Pac. J. Trop. Med. 2016, 9, 402–409. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Peik Lin, T. A minireview on phytochemical and medicinal properties of Clinacanthus nutans. J. Appl. Pharm. Sci. 2020, 11, 15–21. [Google Scholar] [CrossRef]
- Teshima, K.; Kaneko, T.; Ohtani, K.; Kasai, R.; Lhieochaiphant, S.; Picheansoonthon, C.; Yamasaki, K. Sulfur-containing glucosides from Clinacanthus nutans. Phytochemistry 1998, 48, 831–835. [Google Scholar] [CrossRef]
- Gholkar, M.; Li, J.; Daswani, P.; Tetali, P.; Birdi, T. 1H nuclear magnetic resonance-based metabolite profiling of guava leaf extract: An attempt to develop a prototype for standardization of plant extracts. BMC Complement. Med. Ther. 2021, 21, 95. [Google Scholar] [CrossRef]
- Escudero, N.; Marhuenda-Egea, F.; Ibanco-Cañete, R.; Zavala-Gonzalez, E.; Lopez-Llorca, L. A metabolomic approach to study the rhizodeposition in the tritrophic interaction: Tomato, Pochonia chlamydosporia and Meloidogyne javanica. Metabolomics 2014, 10, 788–804. [Google Scholar] [CrossRef] [Green Version]
- Chen, M.; Wang, T.; Jiang, Z.; Shan, C.; Wang, H.; Wu, M.; Wu, M.-J.; Zhang, S.; Zhang, Y.; Zhang, L.-Y. Anti-inflammatory and hepatoprotective effects of total flavonoid C-glycosides from Abrus mollis extracts. Chin. J. Nat. Med. 2014, 12, 590–598. [Google Scholar] [CrossRef]
- Voynow, J.; Shinbashi, M. Neutrophil elastase and chronic lung disease. Biomolecules 2021, 11, 1065. [Google Scholar] [CrossRef]
- Stankovic, M.; Niciforovic, N.; Topuzovic, M.; Solujic, S. Total phenolic content, flavonoid concentrations and antioxidant activity, of the whole plant and Plant Parts Extracts from Teucrium Montanum L. Var. Montanum, F. Supinum (L.) Reichenb. Biotechnol. Biotechnol. Equip. 2011, 25, 2222–2227. [Google Scholar] [CrossRef] [Green Version]
- Shen, Q.; Zhang, B.; Xu, R.; Wang, Y.; Ding, X.; Li, P. Antioxidant activity in vitro of the selenium-contained protein from the Se-enriched Bifidobacterium animalis 01. Anaerobe 2010, 16, 380–386. [Google Scholar] [CrossRef]
- Lee, K.; Oh, Y.; Cho, W.; Ma, J. Antioxidant and anti-inflammatory activity determination of one hundred kinds of pure chemical compounds using offline and online screening HPLC assay. Evid. Based Complement. Altern. Med. 2015, 2015, 165457. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Chew, Y.; Goh, J.; Lim, Y. Assessment of in vitro antioxidant capacity and polyphenolic composition of selected medicinal herbs from Leguminosae family in Peninsular Malaysia. Food Chem. 2009, 116, 13–18. [Google Scholar] [CrossRef]
- Benzie, I.; Strain, J. [2] Ferric reducing/antioxidant power assay: Direct measure of total antioxidant activity of biological fluids and modified version for simultaneous measurement of total antioxidant power and ascorbic acid concentration. Oxid. Antioxid. Part A 1999, 299, 15–27. [Google Scholar] [CrossRef]
- Slamenova, D.; Kozics, K.; Hunakova, L.; Melusova, M.; Navarova, J.; Horvathova, E. Comparison of biological processes induced in HepG2 cells by tert-butyl hydroperoxide (t-BHP) and hydroperoxide (H2O2): The influence of carvacrol. Mutat. Res./Genet. Toxicol. Environ. Mutagenesis 2013, 757, 15–22. [Google Scholar] [CrossRef]
- Wang, H.; Joseph, J. Quantifying cellular oxidative stress by dichlorofluorescein assay using microplate reader. Free Radic. Biol. Med. 1999, 27, 612–616. [Google Scholar] [CrossRef]
Extract | Average TPC (mg GA/g Crude Extract) | Average TFC (mg QE/g Crude Extract) |
---|---|---|
CNL | 21.75 ± 2.41 a | 90.17 ± 0.58 a |
CNS | 6.00 ± 0.43 b | 3.82 ± 0.25 b |
Concentration | ROS Reduction (%) | |
---|---|---|
CNL | CNS | |
13 µg/mL | 7.14 ± 4.163 a | 1.74 ± 2.292 b |
50 µg/mL | 17.44 ± 3.200 a | 9.77 ± 5.485 b |
Metabolites | 1H-NMR Characteristic Signals | |
---|---|---|
CNL | CNS | |
Primary and intermediate metabolites | ||
(1) Valine a | 3.60 (d, J = 11.1 Hz), 2.265 (m), 1.02 (d, J = 6.9 Hz) | 3.61 (d, J = 8.0 Hz), 2.28 (m), 1.02 (d, J = 6.9 Hz) |
(2) Glutamine a | 7.60 (m), 6.88 (s), 3.77 (m), 2.45 (m), 2.11 (m) | 7.60 (m), 6.88 (s), 3.77 (m), 2.45 (m), 2.11 (m) |
(3) Glutamate a | 3.75 (q), 2.35 (m), 2.08 (m) | 3.75 (m), 2.35 (m), 2.08 (m) |
(4) Alanine a | 3.89 (q), 1.45 (d, J = 7.2 Hz) | 3.78 (q), 1.46 (d, J = 7.1 Hz) |
(5) Choline a | 4.08 (m), 3.47 (m), 3.19 (s) | 4.08 (m), 3.47 (m), 3.2 (s) |
(6) Betaine a | 3.87 (s), 3.26 (s) | 3.87 (s), 3.26 (s) |
(7) Proline a | — | 4.12 (t), 3.41 (dt), 3.33 (dt), 2.34 (sext), 2.04 (m), 1.95 (m) |
(8) Anthranilate a | 7.70 (m), 7.29 (t), 6.94 (d, J = 10.9 Hz), 6.86 (t) | — |
(9) α-Glucose b | 5.10 (d, J = 3.7 Hz) | 5.10 (d, J = 3.9 Hz) |
(10) β-Glucose b | — | 4.56 (d, J = 7.9 Hz) |
(11) Fructose b | — | 4.17 (d, J = 7.6 Hz) |
(12) Sucrose b | 5.39 (d, J = 3.8 Hz) | — |
(13) Asparagine b | — | 2.95 (m), 2.77 (m) |
(14) Monoacylmonogalactosyl glycerol b | 5.39 (m), 4.16 (m), 4.01 (m), 3.75 (m), 3.70 (m), 3.52 (m), 2.81 (s), 2.10 (d, J = 18.8 Hz), 2.06 (m), 1.30 (m), 1.29 (s) | 5.36 (m), 4.17 (d, J = 7.6 Hz), 4.02 (m), 3.75 (m), 3.7 (m), 3.47 (m), 3.51 (m), 2.81 (m), 2.11 (m), 2.06 (m), 1.30 (m), 1.28 (s) |
(15) Fatty acid b | 1.31 (m) | 1.36 (m) |
(16) Acetic acid b | 1.95 (s) | 1.95 (s) |
(17) Lactic acid b | 4.09 (m), 1.30 (d, J = 10.0 Hz) | 4.09 (m), 1.32 (m) |
(18) Malonic acid b | 3.08 (s) | 3.10 (s) |
(19) Succinic acid b | 2.51 (s) | — |
(20) Pimelic acid b | 2.10 (d, J = 18.8 Hz), 1.55 (m), 1.30 (m) | 2.11 (m), 1.59 (m), 1.30 (m) |
(21) Ascorbic acid c | 4.47 (d, J = 7.8 Hz), 3.72 (m) | 4.56 (d, J = 7.9 Hz), 3.72 (m) |
Secondary Metabolites | ||
(22) Stigmasterol b | 0.78 (m), 0.82 (m), 0.94 (m), 1.02 (s), 1.06 (d, J = 7.1 Hz), 5.05 (m), 5.16 (m), 5.38 (s) | — |
(23) Lupeol b | — | 4.55(s), 1.68 (s), 1.07 (s), 0.89 (s), 0.86 (s), 0.82 (s), 0.83 (s), 0.75 (s) |
(24) Stigmasterol-β-D-glucoside b | — | 5.10 (d, J = 3.9 Hz), 4.29 (m), 4.9 (m), 4.01 (m), 2.49 (m), 2.11 (m), 2.03 (m), 1.97 (m), 1.87 (m), 1.83 (m), 1.73 (m), 1.71(m), 1.59 (m), 1.54 (m), 1.43 (m), 1.37 (m), 1.26 (m), 1.12 (m), 1.08 (m), 1.03 (m) 1.02 (m), 1.01 (m), 0.97 (m), 0.95 (m), 0.93 (m), 0.91 (m) |
(25) β-Sitosterol b | 5.39 (d, J = 3.8 Hz), 1.02 (s), 0.83 (m), 0.78 (m) | — |
(26) Clinacoside A b | 2.86 (s), 3.81 (m), 4.09 (m), 4.05 (m), 4.70 (d, J = 3.5 Hz), 7.19 (s) | — |
(27) Clinacoside B b | 6.94 (d, J = 10.7 Hz), 4.25 (m), 4.23 (m), 4.01 (d, J = 8.2 Hz), 3.94 (m), 2.51 (s) | — |
(28) Clinacoside C b | 4.05 (m), 3.84 (m), 3.74 (m), 2.65 (s) | 4.04 (m), 3.83 (m), 3.75 (m), 2.67 (s) |
(29) Cycloclinacoside A1 b | 4.74 (m), 4.70 (d, J = 3.5 Hz), 3.49 (d, J = 4.9 Hz), 3.47 (m), 4.09 (m), 4.62 (d, J = 16.8 Hz) | — |
(30) Cycloclinacoside A2 b | 4.70 (d, J = 3.5 Hz), 3.32 (m) | — |
(31) Clinamide A b | 7.32 (d, J = 5.9 Hz), 6.94 (d, J = 10.7 Hz), 3.62 (d, J = 8.4 Hz), 3.47 (m), 3.08 (s) | — |
(32) Clinamide B b | 7.69 (m), 6.65 (d, J = 4.1 Hz), 4.17 (m), 3.55 (m), 2.75 (s), 2.08 (s) | — |
(33) Orientin b | 7.69 (m), 7.47 (m), 7.13 (d), 6.94 (d, J = 10.7 Hz), 6.59 (s), 6.25 (m), 4.09 (m), 3.66 (m), 3.52 (m) | — |
(34) Isoorientin b | 7.32 (d, J = 5.9 Hz), 6.59 (s), 6.52 (s), 3.52 (m), 3.47 (m) | — |
(35) Vitexin b | 8 (d, J = 8.9 Hz), 6.26 (m), 4.88−3.76 (m) | — |
(36) Isovitexin b | 8 (d, J = 8.9 Hz), 6.85 (d, J = 14.8 Hz), 6.84 (s), 6.77(s) | — |
(37) Schaftoside b | 8 (d, J = 8.9 Hz), 6.94 (d, J = 10.7 Hz), 6.77 (s), 3.95–3.21 (m) | — |
(38) Isoschaftoside b | 8 (d, J = 8.9 Hz), 6.94 (d, J = 10.7 Hz), 6.65 (d, J = 4.1 Hz), 4.85 (m), 4.07 (dd, J = 18.3,), 3.94 (m), 3.85 (m), 3.75 (m), 3.64 (m), 3.53 (m), 3.47 (m) | — |
(39) Epigallocatechin b | 7.92 (m), 6.52 (s), 2.70 (m) | — |
(40) 3-O-Methylgallic acid b | 7.19 (s), 3.85 (s) | — |
(41) Catechin b | 3.94 (m), 2.84 (m) | — |
(42) Gallic acid b | 6.98 (s) | 6.95 (s) |
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. |
© 2022 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
Share and Cite
Ng, K.S.; Tan, S.-A.; Bok, C.Y.; Loh, K.E.; Ismail, I.S.; Yue, C.S.; Loke, C.F. Metabolomic Approach for Rapid Identification of Antioxidants in Clinacanthus nutans Leaves with Liver Protective Potential. Molecules 2022, 27, 3650. https://doi.org/10.3390/molecules27123650
Ng KS, Tan S-A, Bok CY, Loh KE, Ismail IS, Yue CS, Loke CF. Metabolomic Approach for Rapid Identification of Antioxidants in Clinacanthus nutans Leaves with Liver Protective Potential. Molecules. 2022; 27(12):3650. https://doi.org/10.3390/molecules27123650
Chicago/Turabian StyleNg, Kai Song, Sheri-Ann Tan, Chui Yin Bok, Khye Er Loh, Intan Safinar Ismail, Chen Son Yue, and Chui Fung Loke. 2022. "Metabolomic Approach for Rapid Identification of Antioxidants in Clinacanthus nutans Leaves with Liver Protective Potential" Molecules 27, no. 12: 3650. https://doi.org/10.3390/molecules27123650