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Multi-analytical characterization of Stevia rebaudiana powder: nutritional, phytochemical, antioxidant, and NMR-based profiling
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  • Published: 25 April 2026

Multi-analytical characterization of Stevia rebaudiana powder: nutritional, phytochemical, antioxidant, and NMR-based profiling

  • Hafiza Fasiha Ahsan1,
  • Shahid Bashir3,
  • Shahnai Basharat2 &
  • …
  • Tabussam Tufail1 

Scientific Reports (2026) Cite this article

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Subjects

  • Biochemistry
  • Chemical biology
  • Chemistry

Abstract

The objective of this study was to characterize the Stevia rebaudiana powder in terms of its nutritional composition, phytochemical potential, antioxidant activity, and NMR-based structural profiling. Proximate analysis indicated that Stevia powder contains considerable amounts of protein (10.52 ± 0.21%) and fiber (7.20 ± 0.69%). The mineral profile revealed the presence of Na, K, Mg, Fe, Zn, Ni and Co, with potassium observed as the most abundant mineral. Phytochemical analysis of ethanolic extract indicated considerable quantities of total phenolic (59.48 ± 0.18 mg GAE/g) and total flavonoid (22.91 ± 0.71 mg QE/g) contents, suggesting the presence of bioactive compounds in Stevia rebaudiana powder. The extract exhibited antioxidant activity with DPPH and ABTS inhibition of 80.51 ± 0.96% and 71.22 ± 0.59%, respectively, compared to the standard BHT. Structural analysis of Stevia rebaudiana powder was performed using nuclear magnetic resonance (1H NMR, 13C NMR, and DEPT experiments) spectroscopy. The 1H NMR spectra indicated the signals corresponding to aliphatic and anomeric proton environments, while the 13C NMR spectra provided qualitative information on CH, CH₂, CH₃, olefinic, and carbonyl carbon environments. DEPT experiments showed qualitative differentiation of methine, methylene, and methyl carbon environments within the steviol glycoside framework. Overall, Stevia rebaudiana powder demonstrates notable nutritional composition and antioxidant activity, along with steviol glycoside related signals suggested by NMR analysis. However, spectral overlap and structural complexity limited the definitive identification of individual compounds based only on one-dimensional NMR. This highlights the need for future studies employing orthogonal analytical techniques including quantitative NMR and complementary spectroscopic methods, for detailed characterization of steviol glycosides and their biological relevance.

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Acknowledgements

The authors gratefully acknowledge the University Institute of Diet and Nutritional Sciences, The University of Lahore, Pakistan.

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Authors and Affiliations

  1. University Institute of Diet and Nutritional Sciences, Faculty of Allied Health Sciences, The University of Lahore, Lahore, Pakistan

    Hafiza Fasiha Ahsan & Tabussam Tufail

  2. Department of Diet and Nutritional Sciences, Rashid Latif Khan University, Lahore, Pakistan

    Shahnai Basharat

  3. University Institute of Food Science and Technology, Faculty of Allied Health Sciences, The University of Lahore, Lahore, Pakistan

    Shahid Bashir

Authors
  1. Hafiza Fasiha Ahsan
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  2. Shahid Bashir
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  3. Shahnai Basharat
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  4. Tabussam Tufail
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Corresponding authors

Correspondence to Hafiza Fasiha Ahsan, Shahid Bashir or Tabussam Tufail.

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The authors declare no competing interests.

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Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by-nc-nd/4.0/.

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Cite this article

Ahsan, H.F., Bashir, S., Basharat, S. et al. Multi-analytical characterization of Stevia rebaudiana powder: nutritional, phytochemical, antioxidant, and NMR-based profiling. Sci Rep (2026). https://doi.org/10.1038/s41598-026-49365-7

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  • Received: 21 March 2026

  • Accepted: 14 April 2026

  • Published: 25 April 2026

  • DOI: https://doi.org/10.1038/s41598-026-49365-7

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Keywords

  • Stevia
  • Nutritional composition
  • 1H NMR
  • 13C NMR
  • DEPT
  • Antioxidant
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