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Plant Science Today (PST)

Research Articles

Vol. 13 No. 3 (2026)

Multivariate profiling reveals extraction-dependent variations in antioxidant and polyphenol content of Citrus maxima (Burm.) Merr. leaves

DOI
https://doi.org/10.14719/pst.13283
Submitted
19 December 2025
Published
23-06-2026 — Updated on 12-08-2026
Versions

Abstract

Citrus maxima (Burm.) Merr. leaves represent a largely underexplored botanical resource with promising antioxidant potential attributed to their polyphenolic constituents. The present study comparatively evaluated the influence of different extraction techniques on the phenolic composition and antioxidant capacity of C. maxima leaf extracts, integrating the obtained data through a multivariate analytical framework. Extracts were prepared using maceration (MAC), ultrasound-assisted extraction (UAE) and microwave-assisted extraction. Total phenolic content (TPC) and total flavonoid content (TFC) were determined and antioxidant activity was evaluated using the 2,2-diphenyl-1-picrylhydrazyl radical-scavenging assay and the 2,2′-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) radical-scavenging assay. Maceration resulted in comparatively higher phenolic and flavonoid contents, whereas UAE demonstrated superior antioxidant performance, as indicated by lower half-maximal inhibitory concentration (IC₅₀) values in both assays. To comprehensively interpret these variations, principal component analysis (PCA) was applied to integrate phenolic and antioxidant parameters. The first principal component accounted for 73.98 % of the total variance, generating a consolidated antioxidant–polyphenol profile that distinctly discriminated among extraction methods. Overall, the findings demonstrate that the antioxidant characteristics of C. maxima leaf extracts are markedly dependent on extraction strategy. The multivariate approach further underscores the utility of principal component analysis for systematically comparing extraction efficiency when multiple interrelated antioxidant indicators are considered.

References

  1. 1. Ignat I, Volf I, Popa VI. A critical review of methods for characterisation of polyphenolic compounds in fruits and vegetables. Food Chem. 2011;126(4):1821–35. https://doi.org/10.1016/j.foodchem.2010.12.026
  2. 2. Ahlawat YK, Singh M, Manorama K, Lakra N, Zaid A, Zulfiqar F. Plant phenolics: neglected secondary metabolites in plant stress tolerance. Braz J Bot. 2024;47(3):703–21. https://doi.org/10.1007/s40415-023-00949-x
  3. 3. Chen X, Li H, Zhang B, Deng Z. The synergistic and antagonistic antioxidant interactions of dietary phytochemical combinations. Crit Rev Food Sci Nutr. 2022;62(20):5658–77. https://doi.org/10.1080/10408398.2021.1888693
  4. 4. Stevenson DE, Lowe T. Plant-derived compounds as antioxidants for health-are they all really antioxidants. Funct Plant Sci Biotechnol. 2009;3(1):1–12.
  5. 5. Maqbool Z, Khalid W, Atiq HT, Shahid M, Ameer K. Citrus waste as a source of bioactive compounds: extraction and utilization in health and food industry. Molecules. 2023;28(4):1636. https://doi.org/10.3390/molecules28041636
  6. 6. Ani PN, Abel HC. Nutrient, phytochemical and antinutrient composition of Citrus maxima fruit juice and peel extract. Food Sci Nutr. 2018;6(3):653–8. https://doi.org/10.1002/fsn3.604
  7. 7. Abudayeh ZH, Al-Khalifa II, Mohammed SM, Ahmad AA. Phytochemical content and antioxidant activities of pomelo peel extract. Pharmacognosy Res. 2019;11(3):244–8. https://doi.org/10.4103/pr.pr_180_18
  8. 8. Akther N, Rahman MA, Hossain MN, Afroze S, Akter F, Johirul IM, et al. In vitro assessment of phytochemicals and antioxidant activity of Citrus maxima leaf extract. J Pharmacogn Phytochem. 2021;10(1):1988–98.
  9. 9. Phetkul U, Thirikorn P, Chaichan K, Mugchanburi S, Paosen S, Voravuthikunchai S. Phytochemical analysis of Citrus grandis (Tubtim siam) leaves and peels extracts and their biological potential. Malays J Fundam Appl Sci. 2025;21(3):2098–110.
  10. 10. Barreca D, Mandalari G, Calderaro A, Smeriglio A, Trombetta D, Felice MR, et al. Citrus flavones: an update on sources, biological functions and health promoting properties. Plants. 2020;9(3):288. https://doi.org/10.3390/plants9030288
  11. 11. Ahmad Z, Rauf A, Orhan IE, Mubarak MS, Akram Z, Islam MR, et al. Antioxidant potential of polyphenolic compounds, sources, extraction, purification and characterization techniques: a focused review. Food Sci Nutr. 2025;13(12):e71259. https://doi.org/10.1002/fsn3.71259
  12. 12. Jolliffe IT, Cadima J. Principal component analysis: a review and recent developments. Philos Trans A Math Phys Eng Sci. 2016;374(2065):20150202. https://doi.org/10.1098/rsta.2015.0202
  13. 13. Osorio-Tobón JF. Recent advances and comparisons of conventional and alternative extraction techniques of phenolic compounds. J Food Sci Technol. 2020;57(12):4299–315. https://doi.org/10.1007/s13197-020-04433-2
  14. 14. Nguyen-Phi N, Le Thi TT, Lam BBP, Nguyen NT, Van HT. Zingiber junceum Gagnep., a new record for the flora of Vietnam and its anatomical and phytochemical characteristics. Trop Nat Hist. 2025;25(1):12–22. https://doi.org/10.58837/tnh.25.1.265505
  15. 15. Mohamed H, Al-Hajhoj M, Al-Saikhan M, Alqahtani N, Zayed M, Moawad M, et al. Green extraction of date palm fruits via ultrasonic-assisted approach: optimizations and antioxidant enrichments. Processes. 2022;10(6):1049. https://doi.org/10.3390/pr10061049
  16. 16. Chamutpong S, Chen CJ, Chaiprateep EO. Optimization ultrasonic–microwave-assisted extraction of phenolic compounds from Clinacanthus nutans using response surface methodology. J Adv Pharm Technol Res. 2021;12(2):190–5. https://doi.org/10.4103/japtr.JAPTR_344_20
  17. 17. Harborne JB. Phytochemical methods: a guide to modern techniques of plant analysis. Springer Sci Bus Media. 1998.
  18. 18. Sukasih E. Total phenol and antioxidant from seed and peel of ripe and unripe of Indonesian sugar apple (Annona squamosa L.) extracted with various solvents. J Pharm. 2015;5(10):20–5.
  19. 19. Baj T, Baryluk A, Sieniawska E. Application of mixture design for optimum antioxidant activity of mixtures of essential oils from Ocimum basilicum L., Origanum majorana L. and Rosmarinus officinalis L. Ind Crops Prod. 2018;115:52–61.
  20. 20. Ozgen M, Reese R, Tulio AZ Jr, Scheerens JC, Miller AR. Modified 2,2-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) method to measure antioxidant capacity of selected small fruits and comparison to ferric reducing antioxidant power (FRAP) and 2,2′-diphenyl-1-picrylhydrazyl (DPPH) methods. J Agric Food Chem. 2006;54(4):1151–7. https://doi.org/10.1021/jf051960d
  21. 21. Sapkota B, Jain V. Evaluation of anti-ulcer activity of Citrus maxima leaves extract in experimental animals. J Clin Exp Pharmacol. 2021;11(2):1–6.
  22. 22. Badve M, Humbare T. Foaming and emulsifying properties of saponin glycosides: a natural non-ionic surfactant. Proc Indian Natl Sci Acad. 2023;89(1):181–8. https://doi.org/10.1007/s43538-023-00151-3
  23. 23. Ganai S, Mukherjee P, Mahali K, Saha A, Hossain A, Soldatov AV, et al. Exploring the solubility and solvation thermodynamics of coumarin in a range of aqua-organic solvents. New J Chem. 2023;47(28):13547–57. https://doi.org/10.1039/D3NJ02412A
  24. 24. Saleh IA, Vinatoru M, Mason TJ, Abdel-Azim NS, Aboutabl EA, Hammouda FM. A possible general mechanism for ultrasound-assisted extraction suggested from the results of UAE of chlorogenic acid from artichoke leaves. Ultrason Sonochem. 2016;31:330–6. https://doi.org/10.1016/j.ultsonch.2016.01.002
  25. 25. Mousavi SA, Nateghi L, Javanmard DM, Ramezan Y, Piravi-Vanak Z. Maceration and ultrasound-assisted methods used for extraction of phenolic compounds and antioxidant activity from Ferulago angulata. J Food Process Preserv. 2022;46(3):e16356. https://doi.org/10.1111/jfpp.16356
  26. 26. Ribeiro AB, Bonafé EG, Silva BC, Montanher PF, Júnior OOS, Boeing JS, et al. Antioxidant capacity, total phenolic content, fatty acids and correlation by principal component analysis of exotic and native fruits from Brazil. J Braz Chem Soc. 2013;24(5):797–804. https://doi.org/10.5935/0103-5053.20130106
  27. 27. Ćujić N, Šavikin K, Janković T, Pljevljakušić D, Zdunić G, Ibrić S. Optimization of polyphenols extraction from dried chokeberry using maceration as traditional technique. Food Chem. 2016;194:135–42. https://doi.org/10.1016/j.foodchem.2015.08.008

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