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Superheated steam extraction and RSM-based optimization of antioxidant activity in Citrus paradisi essential oil and evaluation of antimicrobial activity
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  • Published: 18 January 2026

Superheated steam extraction and RSM-based optimization of antioxidant activity in Citrus paradisi essential oil and evaluation of antimicrobial activity

  • Rameen Waseem1 na1,
  • Muhammad Adnan Ayub1 na1,
  • Muhammad Talha Anjam1,
  • Amjad Hussain2,
  • Ali Bahadur3,4 &
  • …
  • Safura Bibi5 

Scientific Reports , Article number:  (2026) Cite this article

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We are providing an unedited version of this manuscript to give early access to its findings. Before final publication, the manuscript will undergo further editing. Please note there may be errors present which affect the content, and all legal disclaimers apply.

Subjects

  • Analytical chemistry
  • Chemical engineering
  • Green chemistry
  • Plant sciences
  • Software
  • Statistics

Abstract

In this study, response surface methodology (RSM) based on central composite design (CCD) was employed to optimize the antioxidant activity of Citrus paradisi peel essential oil (CPEO) extracted by superheated steam extraction (SHSE). The process parameters temperature (140–160 °C), time (40–80 min), and flow rate (40–80 mL/min) were evaluated. The maximum antioxidant activities, including DPPH-FRSA (95.98%), FRAP (203.87 mg/100 g), and H₂O₂ scavenging (91.38%), were observed at optimum extraction conditions of 150 °C temperature, 60 min time, and 60 mL/min flow rate. Antimicrobial activity results showed that SHSE-extracted essential oil (EO) had better activity compared to hydro-distillation (HD) and steam-distillation (SD). GC–MS analysis revealed that d-limonene, β-pinene, β-myrcene, and trans-carveol were the main components, and their higher concentration in SHSE EOs enhanced their antioxidant and antimicrobial activities. The results indicated that SHSE is an environmentally friendly and effective technique for valorizing Citrus peel waste into high-value bioactive EO. In the future, CPEO may be explored for the development of cost-effective antimicrobial films and pharmaceutical formulations, requiring further compatibility and safety evaluations.

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Data availability

All data generated or analyzed during this study are available from the corresponding author upon reasonable request (mailto: adnanayub@uosahiwal.edu.pk).

Abbreviations

AP:

Adequate precision

ANOVA:

Analysis of variance

CCD:

Central composite design

CPEO:

Citrus paradisi essential oil

CV:

Coefficient of variation

DOE:

Design of experiment

DPPH-FRSA:

DPPH free radical scavenging activity

EO:

Essential oil

FRAP:

Ferric reducing antioxidant power

GC-MS:

Gas chromatography-mass spectrometry

GNB:

Gram negative bacteria

GPB:

Gram positive bacteria

HD:

Hydro-distillation

LSM:

Least square method

MIC:

Minimum inhibitory concentration

RSM:

Response surface technology

SMSS:

Sequential model sum of squares

SD:

Steam distillation

SHSE:

Superheated steam extraction

ZOI:

Zone of inhibition

References

  1. Khalil, M. N., Farghal, H. H. & Farag, M. A. Outgoing and potential trends of composition, health benefits, juice production and waste management of the multi-faceted grapefruit citrus Χ paradisi: A comprehensive review for maximizing its value. Crit. Rev. Food Sci. Nutr. 62 (4), 935–956 (2022).

    Google Scholar 

  2. Maqbool, Z. et al. Citrus waste as source of bioactive compounds: extraction and utilization in health and food industry. Molecules 28 (4), 1636 (2023).

    Google Scholar 

  3. Chaudhary, S. & Singh, B. Grapefruit Peel waste: unlocking the potential for industrial applications in the circular economy. Food Sci. Biotechnol. 34 (10), 2131–2155 (2025).

    Google Scholar 

  4. Venkataraman, S. et al. A comprehensive review on the refinery of citrus Peel towards the production of bioenergy, biochemical and biobased value-added products: present insights and futuristic challenges. Waste Biomass Valoriz. 15 (11), 6491–6512 (2024).

    Google Scholar 

  5. Dosoky, N. S. & Setzer, W. N. Biological activities and safety of Citrus spp. essential oils. Int. J. Mol. Sci. 19 (7), 1966. (2018).

  6. Gupta, V. et al. Pharmacological potentials of citrus paradisi-an overview. Int. J. Phytother Res. 1 (1), 8–17 (2011).

    Google Scholar 

  7. Miya, G. et al. Chemical profiling, toxicity and anti-inflammatory activities of essential oils from three grapefruit cultivars from KwaZulu-Natal in South Africa. Molecules 26 (11), 3387 (2021).

    Google Scholar 

  8. Ali, A. K. et al. Antimicrobial and antioxidant activity of essential oils treated by gamma irradiation extracted from citrus peels. Egypt. J. Chem. 67 (13), 1645–1659 (2024).

    Google Scholar 

  9. Ayub, M. A. et al. Comparison of conventional extraction techniques with superheated steam distillation on chemical characterization and biological activities of syzygium aromaticum L. essential oil. Separations 10 (1), 27 (2023).

    Google Scholar 

  10. Ayub, M. A. et al. Chemical composition, antioxidant, and antimicrobial activities of P. roxburghii Oleoresin essential oils extracted by steam distillation, superheated steam, and supercritical fluid CO2 extraction. J. Food Sci. 88 (6), 2425–2438 (2023).

    Google Scholar 

  11. Aziz, Z. A. et al. Essential oils: extraction techniques, pharmaceutical and therapeutic potential-a review. Curr. Drug Metab. 19 (13), 1100–1110 (2018).

    Google Scholar 

  12. Spadi, A. The application of the steam distillation on different organic matrices: optimization and innovation. (2022).

  13. Rasul, M. G. Conventional extraction methods use in medicinal plants, their advantages and disadvantages. Int. J. Basic. Sci. Appl. Comput. 2, 10–14 (2018).

    Google Scholar 

  14. Dao, T. P. et al. Central composite design, kinetic model, thermodynamics, and chemical composition of pomelo (Citrus Maxima (Burm.) Merr.) essential oil extraction by steam distillation. Processes 9 (11), 2075. (2021).

  15. Dao, T. et al. Kinetic modeling of essential oil hydro-distillation from peels of pomelo (Citrus grandis L.) fruit grown in Southern Vietnam. Sains Malays. 50, 3251–3261 (2021).

    Google Scholar 

  16. Jin, C., Guo, J., Zhu, H. & Wen, J. Optimization of superheated steam treatment conditions for wheat aleurone layer flour. Food Sci. Technol. 42. (2021).

  17. Lee, K. Y. et al. Effect of superheated steam treatment on yield, physicochemical properties and volatile profiles of Perilla seed oil. LWT 135, 110240 (2021).

    Google Scholar 

  18. Lamidi, S. et al. Applications of Response Surface Methodology (RSM) in Product design, development, and Process Optimization (IntechOpen, 2022).

  19. Chen, W. H. et al. A comprehensive review of thermoelectric generation optimization by statistical approach: Taguchi method, analysis of variance (ANOVA), and response surface methodology (RSM). Renew. Sustain. Energy Rev. 169, 112917 (2022).

    Google Scholar 

  20. Halim, N. A. A. et al. Optimization studies and compositional analysis of subcritical water extraction of essential oil from citrus hystrix DC. leaves. J. Supercrit. Fluids. 178, 105384 (2021).

    Google Scholar 

  21. Nguyen, D. H. Application of response surface methodology for the optimization of essential oils from pomelo [citrus grandis (L.) Osbeck] leaves using microwave-assisted hydrodistillation method. Asian J. Chem. 31, 1639–1642 (2019).

    Google Scholar 

  22. Kaanin-Boudraa, G. et al. Response surface methodology and UPLC-QTOF-MSE analysis of phenolic compounds from grapefruit (Citrus paradisi) by-products as novel ingredients for new antioxidant packaging. Lwt 151, 112158 (2021).

    Google Scholar 

  23. Ciğeroğlu, Z. et al. Optimization of ultrasound-assisted extraction of phenolic compounds from grapefruit (Citrus paradisi Macf.) leaves via D‐optimal design and artificial neural network design with categorical and quantitative variables. J. Sci. Food. Agric. 98 (12), 4584–4596 (2018).

    Google Scholar 

  24. El Houda, A. K. N. et al. Chemical composition, antimicrobial and insecticidal activities of citrus paradisi Peel essential oil from Algeria. J. Microbiol. Biotechnol. Food Sci. 9 (6), 1093–1098 (2020).

    Google Scholar 

  25. Czech, A. et al. Mineral content of the pulp and Peel of various citrus fruit cultivars. Biol. Trace Elem. Res. 193 (2), 555–563 (2020).

    Google Scholar 

  26. Ayub, M. A., Hanif, M. A., Sarfraz, R. A. & Shahid, M. Biological activity of Boswellia Serrata Roxb. Oleo gum resin essential oil: effects of extraction by supercritical carbon dioxide and traditional methods. Int. J. Food Prop. 21 (1), 808–820 (2018).

    Google Scholar 

  27. Ayub, M. A. et al. Chemical composition and antimicrobial activity of Boswellia Serrata oleo-gum-resin essential oil extracted by superheated steam. Nat. Prod. Res. 37 (14), 2451–2456 (2023).

    Google Scholar 

  28. Das, A. et al. Antibiofilm and antibacterial activity of essential oil bearing Zingiber officinale Rosc.(Ginger) rhizome against multi-drug resistant isolates. J. Essent. Oil Bearing Plants. 22 (4), 1163–1171 (2019).

    Google Scholar 

  29. Ahmad, M. M., Qureshi, T. M., Nadeem, M. & Asghar, M. Variability in Peel composition and quality evaluation of Peel oils of citrus varieties. J. Agricultural Res. (JAR). 54 (4), 747–756 (2016).

    Google Scholar 

  30. Yuniati, Y., Variyana, Y., Qadariyah, L. & Mahfud, M. The effect of moisture content on essential oil extraction of sweet orange peel (Citrus aurantium L.) using steam distillation method. In AIP Conference Proceedings. (AIP Publishing LLC, 2024).

  31. Suri, S., Singh, A. & Nema, P. K. Recent advances in valorization of citrus fruits processing waste: A way forward towards environmental sustainability. Food Sci. Biotechnol. 30 (13), 1601–1626 (2021).

    Google Scholar 

  32. Bezerra, M. A. et al. Response surface methodology (RSM) as a tool for optimization in analytical chemistry. Talanta 76 (5), 965–977 (2008).

    Google Scholar 

  33. Plaza, M. & Turner, C. Pressurized hot water extraction of bioactives. TRAC Trends Anal. Chem. 71, 39–54 (2015).

    Google Scholar 

  34. Anderson, M. J. & Whitcomb, P. J. RSM Simplified: Optimizing Processes Using Response Surface Methods for Design of Experiments (Productivity Press, 2016).

  35. Zeković, Z. et al. Chemical characterization of polyphenols and volatile fraction of coriander (Coriandrum sativum L.) extracts obtained by subcritical water extraction. Ind. Crops Prod. 87, 54–63 (2016).

    Google Scholar 

  36. Vladić, J. et al. Optimization of satureja Montana subcritical water extraction process and chemical characterization of volatile fraction of extracts. J. Supercrit. Fluids. 120, 86–94 (2017).

    Google Scholar 

  37. Ayub, M. A. et al. Optimizing the extraction of essential oil yield from pistacia lentiscus oleo-gum resin by superheated steam extraction using response surface methodology. Sci. Rep. 14 (1), 25791 (2024).

    Google Scholar 

  38. Mottahedin, P., Haghighi Asl, A. & Khajenoori, M. Extraction of Curcumin and essential oil from curcuma longa L. by subcritical water via response surface methodology. J. Food Process. Preserv. 41 (4), e13095 (2017).

    Google Scholar 

  39. Alboofetileh, M. et al. Subcritical water extraction as an efficient technique to isolate biologically-active fucoidans from Nizamuddinia zanardinii. Int. J. Biol. Macromol. 128, 244–253 (2019).

    Google Scholar 

  40. Jha, A. K. & Sit, N. Comparison of response surface methodology (RSM) and artificial neural network (ANN) modelling for supercritical fluid extraction of phytochemicals from terminalia chebula pulp and optimization using RSM coupled with desirability function (DF) and genetic algorithm (GA) and ANN with GA. Ind. Crops Prod. 170, 113769 (2021).

    Google Scholar 

  41. Mandal, S. C., Mandal, V. & Das, A. K. Essentials of Botanical Extraction: Principles and Applications (Academic Press, 2015).

  42. Cha, J., Kim, C. T., Kim, T. E. & Cho, Y. J. Optimization of subcritical extraction process for cinnamon (Cinnamomum Cassia Blume) using response surface methodology. Food Sci. Biotechnol. 28, 1703–1711 (2019).

    Google Scholar 

  43. Conde-Hernández, L. A. et al. Optimization of extraction of essential oils using response surface methodology: A review. J. Essent. Oil Bearing Plants. 24 (5), 937–982 (2021).

    Google Scholar 

  44. Shahsavarpour, M., Lashkarbolooki, M., Eftekhari, M. J. & Esmaeilzadeh, F. Extraction of essential oils from mentha spicata L.(Labiatae) via optimized supercritical carbon dioxide process. J. Supercrit. Fluids. 130, 253–260 (2017).

    Google Scholar 

  45. Yousefi, M. et al. Supercritical fluid extraction of essential oils. TRAC Trends Anal. Chem. 118, 182–193 (2019).

    Google Scholar 

  46. Doane-Weideman, T. & Liescheski, P. B. Analytical supercritical fluid extraction for food applications. Oil Extr. Anal. 69–99. (2019).

  47. Soh, S. H., Jain, A., Lee, L. Y. & Jayaraman, S. Optimized extraction of Patchouli essential oil from Pogostemon Cablin Benth. With supercritical carbon dioxide. J. Appl. Res. Med. Aromatic Plants. 19, 100272 (2020).

    Google Scholar 

  48. Phewphong, S. et al. Analysis of volatile organic Compounds, Antioxidant, tyrosinase Inhibitory, and antimicrobial activities of essential oils from Citronella grass and Kaffir lime. Trends Sci. 22 (4), 9434–9434 (2025).

    Google Scholar 

  49. Shah, B. B. & Mehta, A. A. In vitro evaluation of antioxidant activity of D-Limonene. Asian J. Pharm. Pharmacol. 4 (6), 883–887 (2018).

    Google Scholar 

  50. Cheng, H. et al. Oxidation characteristics and hazardous of α-pinene, β-pinene and turpentine. Arab. J. Chem. 16 (12), 105322 (2023).

    Google Scholar 

  51. Elgendy, E. M. & Semeih, M. Y. Phyto–Monoterpene Linalool as precursor to synthesis epoxides and hydroperoxides as anti carcinogenic agents via thermal and photo chemical oxidation reactions. Arab. J. Chem. 12 (7), 966–973 (2019).

    Google Scholar 

  52. Mosallaie, F. et al. Unveiling the chemical composition, antioxidant and antibacterial properties, and mechanistic insights of convolvulus arvensis extract through molecular Docking simulations. Appl. Food Res. 4 (2), 100580 (2024).

    Google Scholar 

  53. Noshad, M., Behbahani, B. A. & Nikfarjam, Z. Chemical composition, antibacterial activity and antioxidant activity of citrus Bergamia essential oil: molecular Docking simulations. Food Bioscience. 50, 102123 (2022).

    Google Scholar 

  54. Adeniran, T. R., Oladiji, A. T. & Ezennubia, K. P. Investigation of antioxidant properties in the aqueous extract of apple (Malus pumila). (2025).

  55. Al-Anbari, A. K. H. & Hasan, M. A. Antioxidant activity in some citrus leaves and seeds ethanolic extracts. In International Conference on Advances in Agricultural, Biological and Environmental Sciences (AABES) London (UK). (2015).

  56. Jalil Sarghaleh, S. et al. Evaluation of the constituent compounds, antioxidant, anticancer, and antimicrobial potential of Prangos ferulacea plant extract and its effect on Listeria monocytogenes virulence gene expression. Front. Microbiol. 14, 1202228 (2023).

    Google Scholar 

  57. Sandhu, H. K. et al. Effect of ultrasound-assisted pretreatment on extraction efficiency of essential oil and bioactive compounds from citrus waste by-products. Separations 8 (12), 244 (2021).

    Google Scholar 

  58. Castro-Vazquez, L. et al. Bioactive flavonoids, antioxidant behaviour, and cytoprotective effects of dried grapefruit peels (Citrus paradisi Macf.). Oxid. Med. Cell. Longevity 2016. (2016).

  59. Gargouri, B. et al. Antioxidant capacity and antitumoral activity of citrus paradisi essential oil. Biomedical J. Sci. Tech. Res. 40 (2), 32121–32141 (2021).

    Google Scholar 

  60. Sharif, T. et al. Chemical profiling and in-vitro anti-fungal and antioxidant activities of citrus peel essential oil. Pakistan J. Agric. Sci. 58 (5). (2021).

  61. Torres-Alvarez, C. et al. Chemical composition, antimicrobial, and antioxidant activities of orange essential oil and its concentrated oils. CyTA-Journal Food. 15 (1), 129–135 (2017).

    Google Scholar 

  62. Mohideen, M., Ibrahim, I. F. & Kamaruzaman, N. A. Phytochemical screening and antioxidant properties of essential oil from grapefruit (Citrus paradisi) peel. Asian J. Med. Health Sci. 8 (1). (2025).

  63. Yang, J. & Park, M. J. Antioxidant effects of essential oils from the peels of citrus cultivars. Molecules 30 (4), 833 (2025).

    Google Scholar 

  64. Nourbakhsh, F. et al. From plants to antimicrobials: natural products against bacterial membranes. Phytother. Res. 36 (1), 33–52 (2022).

    Google Scholar 

  65. Lin, H. et al. D-Limonene: promising and sustainable natural bioactive compound. Appl. Sci. 14 (11), 4605 (2024).

    Google Scholar 

  66. Salehi, B. et al. Therapeutic potential of α-and β-pinene: A miracle gift of nature. Biomolecules 9 (11), 738 (2019).

    Google Scholar 

  67. Qiu, Z. et al. High temperature induced disruption of the cell wall integrity and structure in pleurotus ostreatus mycelia. Appl. Microbiol. Biotechnol. 102 (15), 6627–6636 (2018).

    Google Scholar 

  68. Maurya, A., Prasad, J., Das, S. & Dwivedy, A. K. Essential oils and their application in food safety. Front. Sustainable Food Syst. 5, 653420 (2021).

    Google Scholar 

  69. Garde, S., Chodisetti, P. K. & Reddy, M. Peptidoglycan: structure, synthesis, and regulation. EcoSal Plus 9 (2). (2021).

  70. Tavares, T. D. et al. Activity of specialized biomolecules against gram-positive and gram-negative bacteria. Antibiotics 9 (6), 314 (2020).

    Google Scholar 

  71. Ahmed, A. O. Comparative Study of Antimicrobial Activities and GC-MS Analysis of Bioactive Compounds of Essential Oil of Selected Citrus Peels (Kwara State University (Nigeria), 2019).

  72. Mohammed, I. O., Alrasheid, A. A. & Hussein Ayoub, S. M. GC-MS analysis and study of the antimicrobial activity of citrus paradisi, citrus aurantifolia, and citrus sinensis Peel essential oils as hand sanitizer. Int. J. Microbiol. 2024 (1), 4957712 (2024).

    Google Scholar 

  73. Javed, S. et al. Chemical constituents, antimicrobial and antioxidant activity of essential oil of citrus Limetta var. Mitha (sweet lime) Peel in Pakistan. Afr. J. Microbiol. Res. 7 (24), 3071–3077 (2013).

    Google Scholar 

  74. Denkova-Kostova, R. et al. Chemical composition, antioxidant and antimicrobial activity of essential oils from Tangerine (Citrus reticulata L.), grapefruit (Citrus paradisi L.), lemon (Citrus lemon L.) and cinnamon (Cinnamomum zeylanicum Blume). Z. für Naturforschung C. 76 (5–6), 175–185 (2021).

    Google Scholar 

  75. Behbahani, B. A. et al. Synergistic activity of satureja intermedia and Ducrosia anethifolia essential oils and their interaction against foodborne pathogens: A multi-ligand molecular Docking simulation. Lwt 205, 116487 (2024).

    Google Scholar 

  76. Noshad, M., Behbahani, B. A., Nikfarjam, Z. & Zargari, F. Antimicrobial activity between coriandrum sativum seed and cuminum cyminum essential oils against foodborne pathogens: A multi-ligand molecular Docking simulation. Lwt 185, 115217 (2023).

    Google Scholar 

  77. Behbahani, B. A. et al. First report on the synergy of Nepeta menthoides and Nepeta cephalotes essential oils for antimicrobial and preservation applications: a multi-ligand molecular Docking simulation. Appl. Food Res. 5 (1), 100707 (2025).

    Google Scholar 

  78. Andrade-Ochoa, S. et al. Differential antimicrobial effect of essential oils and their main components: insights based on the cell membrane and external structure. Membranes 11 (6), 405 (2021).

    Google Scholar 

  79. Khorshidian, N., Yousefi, M., Khanniri, E. & Mortazavian, A. M. Potential application of essential oils as antimicrobial preservatives in cheese. Innovative Food Sci. Emerg. Technol. 45, 62–72 (2018).

    Google Scholar 

  80. Anwar, T. et al. Citrus sinensis Peel oil extraction and evaluation as an antibacterial and antifungal agent. Microorganisms 11 (7), 1662 (2023).

    Google Scholar 

  81. Elgat, W. A. A. et al. Eucalyptus camaldulensis, citrus aurantium, and citrus sinensis essential oils as antifungal activity against Aspergillus flavus, Aspergillus niger, Aspergillus terreus, and fusarium culmorum. Processes 8 (8), 1003 (2020).

    Google Scholar 

  82. Han, Y., Sun, Z. & Chen, W. Antimicrobial susceptibility and antibacterial mechanism of limonene against Listeria monocytogenes. Molecules 25 (1), 33 (2019).

    Google Scholar 

  83. Plaza, M. & Marina, M. L. Pressurized hot water extraction of bioactives. TRAC Trends Anal. Chem. 117201. (2023).

  84. Zhu, Y., Li, C., Cui, H. & Lin, L. Encapsulation strategies to enhance the antibacterial properties of essential oils in food system. Food Control. 123, 107856 (2021).

    Google Scholar 

  85. Raj, B., John, S., Chandrakala, V. & Kumari, G. H. Green extraction techniques for phytoconstituents from natural products. In Medicinal Plants. (IntechOpen, 2022).

  86. Paoli, M. et al. Chemical composition of the leaf essential oil of grapefruits (Citrus paradisi Macf.) in relation with the genetic origin. J. Essent. Oil Res. 28 (4), 265–271 (2016).

    Google Scholar 

  87. Ndao, A. & Adjallé, K. Overview of the biotransformation of limonene and α-pinene from wood and citrus residues by microorganisms. In Waste (MDPI, 2023).

  88. Sell, C. S. A fragrant introduction to terpenoid chemistry. Royal Soc. Chem., (2019).

  89. Charbonneau, L., Foster, X., Zhao, D. & Kaliaguine, S. Catalyst-free epoxidation of limonene to limonene dioxide. ACS Sustain. Chem. Eng. 6 (4), 5115–5121 (2018).

    Google Scholar 

  90. de Groot, A. Limonene hydroperoxides. Dermatitis 30 (6), 331–335 (2019).

    Google Scholar 

  91. Hegazy, M. M. et al. Essential oils: the science of extraction and its implications for composition and biological activity—a review. Food Anal. Methods 1–31 (2025).

  92. Cox-Georgian, D., Ramadoss, N., Dona, C. & Basu, C. Therapeutic and medicinal uses of terpenes. Med. Plants Farm Pharm. 333–359. (2019).

  93. Li, C. et al. Variation in compositions and biological activities of essential oils from four citrus species: Citrus limon, Citrus sinensis, Citrus paradisi, and Citrus reticulata. Chemi. Biodiversity 19 (4), e202100910 (2022).

  94. Li, Y. et al. The chemical composition and antibacterial and antioxidant activities of five citrus essential oils. Molecules 27 (20), 7044 (2022).

    Google Scholar 

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Acknowledgements

We would like to express our gratitude to Dr. Fahim Arshad, Associate Professor, Department of botany, University of Okara, Pakistan for his invaluable assistance in verification of plant material.

Funding

The authors would like to acknowledge the support from the Higher Education Commission (HEC), Islamabad, Pakistan, for funding Project Number 20-15988/NRPU/R&D/HEC/2021, International Collaborative Research Program (ICRP2023008), Internal Faculty/Staff Research Support Programs (IRSPC2024007) at Wenzhou Kean University, China and Wenzhou Association for Science and Technology- Service and Technology Innovation Program (No. KJFW2024-054).

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Author notes

  1. Rameen Waseem and Muhammad Adnan Ayub contributed equally to this work.

Authors and Affiliations

  1. Department of Chemistry, University of Sahiwal, Sahiwal, 57000, Pakistan

    Rameen Waseem, Muhammad Adnan Ayub & Muhammad Talha Anjam

  2. Department of Chemistry, University of Okara, Okara, 56300, Pakistan

    Amjad Hussain

  3. Department of Chemistry, College of Science, Mathematics, and Technology, Wenzhou-Kean University, Wenzhou, 325060, China

    Ali Bahadur

  4. Dorothy and George Hennings College of Science, Mathematics and Technology, Kean University, 1000 Morris Ave, Union, NJ, 07083, USA

    Ali Bahadur

  5. Department of Botany, University of Agriculture, Faisalabad, 38040, Pakistan

    Safura Bibi

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  1. Rameen Waseem
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  2. Muhammad Adnan Ayub
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Contributions

Rameen Waseem: Investigation, Writing—original draft, Formal analysis, Writing—review & editing. Muhammad Adnan Ayub: Conceptualization, Writing—review & editing, project administration, supervision. Muhammad Talha Anjam: Investigation, Writing—original draft. Amjad Hussain: Methodology, resources. Ali Bahadur: Software, Writing—review & editing. Safura Bibi: Methodology, validation.

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Correspondence to Muhammad Adnan Ayub or Ali Bahadur.

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Waseem, R., Ayub, M.A., Anjam, M.T. et al. Superheated steam extraction and RSM-based optimization of antioxidant activity in Citrus paradisi essential oil and evaluation of antimicrobial activity. Sci Rep (2026). https://doi.org/10.1038/s41598-026-35099-z

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  • Received: 17 February 2025

  • Accepted: 02 January 2026

  • Published: 18 January 2026

  • DOI: https://doi.org/10.1038/s41598-026-35099-z

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Keywords

  • Essential oil
  • Antimicrobial activity
  • Antioxidant activity
  • Chemical composition
  • Optimization
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Scientific Reports (Sci Rep)

ISSN 2045-2322 (online)

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