Valorization of Cultivated and Wild Spinach Varieties: Comparative Phytochemical Screening and Evaluation of Antioxidant and Anti-Inflammatory Activities
DOI:
https://doi.org/10.22399/ijcesen.5282Keywords:
Chenopodium bonus-henricus, Spinacia oleracea, TLC, FRAP, DPPH, IC50Abstract
This study aims to compare the cultivated spinach Spinacia oleracea and wild spinach Chenopodium bonus-henricus through detailed phytochemical screening and evaluation of their biological activities. Quantitative analysis shows that C. bonus-henricus leaves have the highest polyphenol (102.9 ± 6.8 mg GAE/g) and flavonoid (74.3 ± 4.7 mg QE/g) contents. Screening by thin-layer chromatography (TLC) reveals greater molecular diversity in leaf tissues, with the identification of flavonoids, coumarins, and the exclusive presence of terpenoids and quinones. On the biological level, the wild species consistently outperforms the cultivated variety, displaying an antioxidant capacity of 486.5 ± 14.1 µmol TE/g (FRAP assay) and a stronger radical scavenging activity with a lower DPPH IC₅₀ value (37 µg/mL) compared to the cultivated spinach (43.65 µg/mL). Similarly, anti-inflammatory activity assessment via bovine serum albumin (BSA) denaturation inhibition revealed a lower IC₅₀ for C. bonus-henricus leaves (216.1 µg/mL) relative to S. oleracea (416.1 µg/mL), reaching 81.2% inhibition at 800 µg/mL. These findings confirm that tissue compartmentalization and wild origin promote the accumulation of protective secondary metabolites, positioning C. bonus-henricus as a promising bioactive resource for nutritional and pharmaceutical applications.
References
1. Alruways, S. K., Alenazy, S. R., Zainy, F. S. R., Alghamdi, A. M. S., Althomali, M. M. B., Khairallah, W. S. I., Al Otaibi, N. M. Z., Ageel, F. M., Jan, R. F. A. K., Ahmed, G. A. M. S., & Almagnoni, A. S. A. (2024). Biomarkers of oxidative stress in chronic diseases: Clinical and laboratory perspectives. The Review of Diabetic Studies, 20, 290–296. https://doi.org/10.70082/5a85z086
2. Agati, G., Brunetti, C., Fini, A., Gori, A., Guidi, L., Landi, M., Sebastiani, F., & Tattini, M. (2020). Are Flavonoids Effective Antioxidants in Plants? Twenty Years of Our Investigation. Antioxidants, 9(11), 1098. https://doi.org/10.3390/antiox9111098
3. Akash, M. S. H., & Rehman, K. (2020). Thin layer chromatography. In Essentials of pharmaceutical analysis (Chapter 12). Springer. https://doi.org/10.1007/978-981-15-1547-7_12
4. Badmus, U. O., Crestani, G., Cunningham, N., Havaux, M., Urban, O., & Jansen, M. A. K. (2022). UV Radiation Induces Specific Changes in the Carotenoid Profile of Arabidopsis thaliana. Biomolecules, 12(12), 1879. https://doi.org/10.3390/biom12121879
5. Batool, Z., Lone, J. F., Singh, K., & Gairola, S. (2024). Nutraceutical and antioxidant potential of selected wild edible plants from the cold arid desert of Ladakh, India. Notulae Botanicae Horti Agrobotanici Cluj Napoca, 52(1), Article 13286. https://doi.org/10.15835/nbha52113286
6. Belew, A. A., Hanan, G. G. M. W., Meshesha, D. S., et al. (2025). Evaluation of total phenolic, flavonoid contents, antioxidant and antibacterial activity of leaf extracts from Rhus vulgaris. Discover Plants, 2, 141. https://doi.org/10.1007/s44372-025-00222-3
7. Benzie, I. F. F., & Strain, J. J. (1996). The ferric reducing ability of plasma (FRAP) as a measure of “antioxidant power”: The FRAP assay. Analytical Biochemistry, 239(1), 70–76. https://doi.org/10.1006/abio.1996.0292
8. Bolat, E., Sarıtaş, S., Duman, H., Eker, F., Akdaşçi, E., Karav, S., & Witkowska, A. M. (2024). Polyphenols: Secondary Metabolites with a Biological Impression. Nutrients, 16(15), 2550. https://doi.org/10.3390/nu16152550
9. Brand-Williams, W., Cuvelier, M. E., & Berset, C. (1995). Use of a free radical method to evaluate antioxidant activity. LWT – Food Science and Technology, 28(1), 25–30. https://doi.org/10.1016/S0023-6438(95)80008-5
10. Bruneton, J. (2016). Pharmacognosie: Phytochimie, plantes médicinales (5e éd.). Lavoisier Tec & Doc.
11. Chatepa, L. E. C., Mwamatope, B., Chikowe, I., & Masamba, K. G. (2024). Effects of solvent extraction on the phytoconstituents and in vitro antioxidant activity properties of leaf extracts of the two selected medicinal plants from Malawi. BMC Complementary Medicine and Therapies, 24, 317. https://doi.org/10.1186/s12906-024-04619-7
12. Chong, Z. Z., & Souayah, N. (2025). Oxidative Stress: Pathological Driver in Chronic Neurodegenerative Diseases. Antioxidants, 14(6), 696. https://doi.org/10.3390/antiox14060696
13. Dawson, R. M. C., Elliott, D. C., Elliott, W. H., & Jones, K. M. (1991). Data for biochemical research (3rd ed.). Oxford University Press.
14. Dong, M., Ming, X., Xiang, T., Feng, N., Zhang, M., Ye, X., He, Y., & Zhou, M. (2024). Recent research on the physicochemical properties and biological activities of quinones and their practical applications: a comprehensive review. Food & Function, 15(18), 8973–8997. https://doi.org/10.1039/D4FO02600D
15. Eligar, S. M. (2022). In vitro lipoxygenase and hemolysis inhibition by polyphenolic antioxidants from tropical green leafy vegetables. Emirates Journal of Food & Agriculture (EJFA), 34(7). https://doi.org/10.9755/ejfa.2022.v34.i7.2897
16. Gülçin, İ. (2020). Antioxidants and antioxidant methods: An updated overview. Archives of Toxicology, 94, 651–715. https://doi.org/10.1007/s00204-020-02689-3
17. Faujan, N., Zubairi, S., & Baker, A. (2023). Nutritional and bioactive constituents of antioxidant and antimicrobial properties in Spinacia oleracea: A review. Sains Malaysiana, 52(9), 2571–2585. https://doi.org/10.17576/jsm-2023-5209-08
18. Fu, L., Sun, Y., Ding, L., Wang, Y., Gao, Z., Wu, Z., Wang, S., Li, W., & Bi, Y. (2016). Mechanism evaluation of the interactions between flavonoids and bovine serum albumin based on multi-spectroscopy, molecular docking and Q-TOF HR-MS analyses. Food Chemistry, 203, 150–157. https://doi.org/10.1016/j.foodchem.2016.01.105
19. Hameed, K., Khan, M. S., Fatima, A., Shah, S. M., & Abdullah, M. A. (2023). Exploring the world of thin-layer chromatography: A review. Asian Journal of Applied Chemistry Research, 14(3), 23–38. https://doi.org/10.9734/ajacr/2023/v14i3268
20. Harborne, A. J. (1998). Phytochemical methods: A guide to modern techniques of plant analysis (5th ed.). Springer Science & Business Media.
21. Hassanpour, S., & Doroudi, A. (2023). Review of the antioxidant potential of flavonoids as a subgroup of polyphenols and partial substitute for synthetic antioxidants. Avicenna Journal of Phytomedicine, 13, 354 - 376. https://doi.org/10.22038/ajp.2023.21774
22. Jalouli, M., Rahman, M. A., Biswas, P., Rahman, H., Harrath, A. H., Lee, I. S., Kang, S., Choi, J., Park, M. N., & Kim, B. (2025). Targeting natural antioxidant polyphenols to protect neuroinflammation and neurodegenerative diseases: a comprehensive review. Frontiers in pharmacology, 16, 1492517. https://doi.org/10.3389/fphar.2025.1492517
23. Jaiswal, S. G., Patel, M., Saxena, D. K., & Naik, S. N. (2017). Comparison of measurements of antioxidant activity in the selected leafy vegetables depending on extraction solvent. Journal of Horticultural Research, 25(2), 75–80. https://doi.org/10.1515/johr-2017-0023
24. Jomova, K., Raptova, R., Alomar, S. Y., Alwasel, S. H., Nepovimova, E., Kuca, K., & Valko, M. (2023). Reactive oxygen species, toxicity, oxidative stress, and antioxidants: chronic diseases and aging. Archives of Toxicology, 97(10), 2499–2574. https://doi.org/10.1007/s00204-023-03562-9
25. Kabir, M. F., Mohd Ali, J., Abolmaesoomi, M., et al. (2017). Melicope ptelefolia leaf extracts exhibit antioxidant activity and exert anti-proliferative effect with apoptosis induction on four different cancer cell lines. BMC Complementary and Alternative Medicine, 17, 252. https://doi.org/10.1186/s12906-017-1761-9
26. Kazi, Z., Singh, S., & Lobo, V. (2025). Phytochemical evaluation and in vitro antioxidant potential of Cycas circinalis L. Current Botany, 16, 59–67. https://doi.org/10.25081/cb.2025.v16.9123
27. Kokanova-Nedialkova, Z., Nedialkov, P., Kondeva-Burdina, M., Simeonova, R., Tzankova, V., & Aluani, D. (2017). Chenopodium bonus-henricus L. – A source of hepatoprotective flavonoids. Fitoterapia, 118, 13–20. https://doi.org/10.1016/j.fitote.2017.02.001
28. Kokanova-Nedialkova, Z., & Nedialkov, P. (2021). Validated UHPLC-HRMS method for simultaneous quantification of flavonoid contents in the aerial parts of Chenopodium bonus-henricus L. (wild spinach). Pharmacia, 68, 597–601. https://doi.org/10.3897/pharmacia.68.e69781
29. Kolar, F. R., Kambhar, S. V., Kamble, V. S., & Daphedar, A. B. (2024). Comparative examination of phenolic content, flavonoid content, and antioxidant efficacy of Chenopodium album L. and Chenopodium pumilio R. Br. İstanbul Journal of Pharmacy, 54(2), 165–174. https://doi.org/10.26650/istanbuljpharm.2024.1313743
30. Kondeva-Burdina, M., Panayotova, D., Nedialkov, P. T., & Kokanova-Nedialkova, Z. (2025). Flavonoids and saponins from two Chenopodium species (C. foliosum Asch. and C. bonus-henricus L.) Preliminary evaluation for hMAO-A/B, neuroprotective activity, and validated UHPLC-HRMS quantification of ethanolic extract from C. foliosum. Molecules, 30(5), 1061. https://doi.org/10.3390/molecules30051061
31. Lingwan, M., Pradhan, A. A., Kushwaha, A. K., Dar, M. A., Bhagavatula, L., & Datta, S. (2023). Photoprotective role of plant secondary metabolites: Biosynthesis, photoregulation, and prospects of metabolic engineering for enhanced protection under excessive light. Environmental and Experimental Botany, 209, 105300. https://doi.org/10.1016/j.envexpbot.2023.105300
32. Litvinova, L. S. (2008). Determination of the retention characteristics in thin-layer chromatography taking into account the actual position of the mobile phase front. Russian Journal of Applied Chemistry, 81, 1538–1542. https://doi.org/10.1134/S1070427208090127
33. Mammen, D., & Daniel, M. (2012). A critical evaluation on the reliability of two aluminum chloride chelation methods for quantification of flavonoids. Food Chemistry, 135(3), 1365–1368. https://doi.org/10.1016/j.foodchem.2012.05.109
34. Masiala, A., Vingadassalon, A., & Aurore, G. (2023). Comparative studies on polyphenolic composition, antioxidant and antimicrobial activities of Artocarpus altilis (L.) leaves. In Proceedings of the 4th International Conference on Food Bioactives & Health (Vol. 15, No. 13, pp. 6847–6882). https://doi.org/10.1039/d4fo00509k
35. Mazzucotelli, C. A., González‐Aguilar, G. A., Villegas‐Ochoa, M. A., Domínguez‐Avila, A. J., Ansorena, M. R., & Di Scala, K. C. (2018). Chemical characterization and functional properties of selected leafy vegetables for innovative mixed salads. Journal of Food Biochemistry, 42(1), e12461. https://doi.org/10.1111/jfbc.12461
36. Mishra, M. K., Pandey, S., Niranjan, A., et al. (2021). Comparative analysis of phenolic compounds from wild and in vitro propagated plant Thalictrum foliolosum and antioxidant activity of various crude extracts. Chemical Papers, 75, 4873–4885. https://doi.org/10.1007/s11696-021-01708-6
37. Mizushima, Y., & Kobayashi, M. (1968). Interaction of anti-inflammatory drugs with serum proteins, especially with some biologically active proteins. Journal of Pharmacy and Pharmacology, 20(3), 169–173. https://doi.org/10.1111/j.2042-7158.1968.tb09718.x
38. Munteanu, I. G., & Apetrei, C. (2021). Analytical methods used in determining antioxidant activity: A review. International Journal of Molecular Sciences, 22(7), Article 3380. https://doi.org/10.3390/ijms22073380
39. Mroczek, A. (2015). Phytochemistry and bioactivity of triterpene saponins from Amaranthaceae family. Phytochemistry Reviews, 14, 577–605. https://doi.org/10.1007/s11101-015-9394-4
40. Nani, A., Murtaza, B., Sayed Khan, A., Khan, N. A., & Hichami, A. (2021). Antioxidant and Anti-Inflammatory Potential of Polyphenols Contained in Mediterranean Diet in Obesity: Molecular Mechanisms. Molecules, 26(4), 985. https://doi.org/10.3390/molecules26040985
41. Panche, A. N., Diwan, A. D., & Chandra, S. R. (2016). Flavonoids: An overview. Journal of Nutritional Science, 5, e47. https://doi.org/10.1017/jns.2016.41
42. Pandey, S., Singh, D., Yadav, J. P., & Verma, A. (2025). Medicinal potential of Curcuma caesia Roxb.: Phytochemical composition and TLC profile of hydroalcoholic rhizome extract. Journal of Drug Delivery and Therapeutics, 15(6), 41–47. https://doi.org/10.22270/jddt.v15i6.7181
43. Reddy, V. P. (2023). Oxidative Stress in Health and Disease. Biomedicines, 11(11), 2925. https://doi.org/10.3390/biomedicines11112925
44. Roberts, J., & Moreau, R. (2016). Functional properties of spinach (Spinacia oleracea L.) phytochemicals and bioactives. Food & Function, 7(8), 3337–3353. https://doi.org/10.1039/c6fo00051g
45. Rudrapal, M., Khairnar, S. J., Khan, J., Dukhyil, A. B., Ansari, M. A., Alomary, M. N., Alshabrmi, F. M., Palai, S., Deb, P. K., & Devi, R. (2022). Dietary polyphenols and their role in oxidative stress-induced human diseases: Insights into protective effects, antioxidant potentials and mechanism(s) of action. Frontiers in Pharmacology, 13, 806470. https://doi.org/10.3389/fphar.2022.806470
46. Sasidharan, S., Chen, Y., Saravanan, D., Sundram, K. M., & Latha, L. Y. (2011). Extraction, isolation and characterization of bioactive compounds from plants’ extracts. African Journal of Traditional, Complementary and Alternative Medicines, 8(1), 1–10. https://doi.org/10.4314/ajtcam.v8i1.60483
47. Shrivastava, A. K., Keshari, M., Neupane, M., Chaudhary, S., Dhakal, P. K., Shrestha, L., Gurung, S., Rai, R. B., Adhikari, B., & Yadav, R. K. (2023). Evaluation of antioxidant and anti‐inflammatory activities, and metabolite profiling of selected medicinal plants of Nepal. Journal of Tropical Medicine, 2023(1), Article 6641018. https://doi.org/10.1155/2023/6641018
48. Singleton, V. L., & Rossi, J. A., Jr. (1965). Colorimetry of total phenolics with phosphomolybdic–phosphotungstic acid reagents. American Journal of Enology and Viticulture, 16(3), 144–158. https://doi.org/10.5344/ajev.1965.16.3.144
49. Tholl, D. (2015). Biosynthesis and biological functions of terpenoids in plants. Advances in Biochemical Engineering/Biotechnology, 148, 63–106. https://doi.org/10.1007/10_2014_295
50. Trease, G. E., & Evans, W. C. (1987). Pharmacognosy (13th ed.). Baillière Tindall.
51. Tretter, V., Hochreiter, B., Zach, M. L., Krenn, K., & Klein, K. U. (2022). Understanding cellular redox homeostasis: A challenge for precision medicine. International Journal of Molecular Sciences, 23(1), 106. https://doi.org/10.3390/ijms23010106
52. Venugopala, K. N., Rashmi, V., & Odhav, B. (2013). Review on natural coumarin lead compounds for their pharmacological activity. BioMed Research International, 2013, 963248. https://doi.org/10.1155/2013/963248
53. Wagner, H., Bladt, S., & Zgainski, E. M. (1996). Screening of unknown commercial drugs. In Plant drug analysis (Chapter 19). Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-00574-9_19
54. Wang, X., Ma, H., Zhou, X., Chen, Q., & Yang, Y. (2021). UV B treatment enhances phenolic acid accumulation and antioxidant capacity of barley seedlings. LWT – Food Science and Technology, 144, 112445. https://doi.org/10.1016/j.lwt.2021.112445
55. Weletnsae, T., Tadege, T., Hintsa, K., Hagos, Z., Babu, D. S., Palieti, J. D., & Chaithanya, K. K. (2024). In vitro screening of 5-LOX/COX-2 dual inhibitors from selected medicinal plant Maytenus arbutifolia. Research Journal of Pharmacy and Technology, 17(1), 241–248. https://doi.org/10.52711/0974-360X.2024.00038
56. Yang, J., Guo, J., & Yuan, J. (2008). In vitro antioxidant properties of rutin. LWT–Food Science and Technology, 41(6), 1060–1066. https://doi.org/10.1016/j.lwt.2007.06.010
57. Zagoskina, N., Zubova, M., Nechaeva, T., Kazantseva, V., Goncharuk, E., Katanskaya, V., Baranova, E., & Aksenova, M. (2023). Polyphenols in Plants: Structure, Biosynthesis, Abiotic Stress Regulation, and Practical Applications (Review). International Journal of Molecular Sciences, 24. https://doi.org/10.3390/ijms241813874.
58. Zhong, R., Cui, D., & Ye, Z. (2019). Secondary cell wall biosynthesis. The New Phytologist, 221(4), 1703–1723. https://doi.org/10.1111/nph.15537
59. Zhang, H., He, H., Song, W., & Zheng, L. (2023). Pre-harvest UVB irradiation enhances the phenolic and flavonoid content, and antioxidant activity of green- and red-leaf lettuce cultivars. Horticulturae, 9(6), 695. https://doi.org/10.3390/horticulturae906695
60. Zhang, Y., Huang, W., Zhang, C., Huang, H., Yang, S., Wang, Y., Huang, Z., Tang, Y., Li, X., Lian, H., Li, H., Zhang, F., & Sun, B. (2023). Variation in the Main Health-Promoting Compounds and Antioxidant Capacity of Three Leafy Vegetables in Southwest China. Molecules, 28(12), 4780. https://doi.org/10.3390/molecules28124780.
61. Zheng, Y., Deng, G., & Zhang, Y. (2021). Multiple free radical scavenging reactions of flavonoids. Dyes and Pigments, 198, 109877. https://doi.org/10.1016/j.dyepig.2021.109877
62. Zhishen, J., Mengcheng, T., & Jianming, W. (1999). The determination of flavonoid contents in mulberry and their scavenging effects on superoxide radicals. Food Chemistry, 64(4), 555–559. https://doi.org/10.1016/S0308-8146(98)00102-2
Downloads
Published
How to Cite
Issue
Section
License
Copyright (c) 2026 International Journal of Computational and Experimental Science and Engineering
This work is licensed under a Creative Commons Attribution 4.0 International License.
