PHYTOCHEMICAL SCREENING, PHARMACOGNOSTICS CH- ARACTERIZATION, AND CHROMATOGRAPHIC ANALYSIS (TLC/HPTLC) OF STEVIA REBAUDIANA BERTONI LEAF EXTRACT

  • Jay Prakash Singh Lords University, Alwar Rajsthan-301001
  • Shikha Sharma Lords University, Alwar Rajsthan-301001

Abstract

The growing demand for natural sweeteners as alternatives to synthetic sugars has heightened interest in Stevia rebaudiana Bertoni, a perennial shrub native to South America, known for its high-intensity, non-caloric steviol glycosides. This study aimed to evaluate the pharmacognostic, phytochemical, and chromatographic properties of S. rebaudiana leaves to establish quality control parameters and validate its therapeutic potential. Fresh leaves were authenticated, shade-dried, and subjected to ethanolic extraction via cold maceration. Macroscopic and microscopic analyses revealed characteristic features such as lanceolate leaves with serrated margins, anisocytic stomata, and calcium oxalate crystals. Physicochemical evaluation showed acceptable moisture content (6.2%), total ash (9.4%), and extractive values (14.3% water-soluble). Phytochemical screening confirmed the presence of steviol glycosides, flavonoids, phenolics, and terpenoids, supporting its antioxidant and antidiabetic properties. Chromatographic profiling using TLC and HPTLC resolved key markers, with stevioside identified at Rf 0.67 (TLC) and 0.65 (HPTLC), providing a reliable fingerprint for standardization. The findings underscore S. rebaudiana's potential as a natural sweetener and therapeutic agent, while establishing quality benchmarks for its use in nutraceutical and pharmaceutical applications. Further clinical studies are recommended to explore its efficacy and safety.

Keywords: Stevia rebaudiana, pharmacognostic evaluation, phytochemical screening, HPTLC fingerprinting, steviol glycosides, natural sweetener

References

An HJ, & Jin H. Physicochemical standardization of herbal materials: Methods and applications. Journal of Pharmacognosy and Phytochemistry. 2012;1(3):1-12.
2. Ceunen S, & Geuns JM. Steviol glycosides: Chemical diversity, metabolism, and function. Journal of Natural Products. 2013;76(6):1201-1228. https://doi.org/10.10 21/np400203b
3. Chatsudthipong V, & Muanprasat C. Therapeutics, Stevioside and related compounds: Therapeutic benefits beyond sweetness. Pharmacology & Therapeutics. 2009;121(1):41-54. https://doi.org/10.10 16/j.pharmthera.2008.09.007
4. Geuns JM. Sativoside. Phytochemistry. 2003;64(5):913-921. https://doi.org/10.1016/S0031-9422(03)00426-6
5. Goyal SK, Samsher S, & Goyal RK. Stevia (Stevia rebaudiana) a bio-sweetener: A review. International Journal of Food Sciences and Nutrition. 2010;61(1):1-10. https://doi.org/10.3109/09637480903193049
6. Kaushik R, Narayanan P, Vasudevan V, Muthukumaran G, & Antony U. Nutrient composition of cultivated stevia leaves and the influence of polyphenols and plant pigments on sensory and antioxidant properties of leaf extracts. Journal of Food Science and Technology. 2010;47(1):27-33. https://doi. org/10.1007/s13197-010-0011-7
7. Lemus-Mondaca R, Vega-Gálvez A, Zura-Bravo L, & Ah-Hen K. Stevia rebaudiana Bertoni, source of a high-potency natural sweetener: A comprehensive review on the biochemical, nutritional, and functional aspects. Food Chemistry. 2012;132(3):1121-1132. https://doi.org/10 .1016/j.foodchem.2011.11.140
8. Griffiths CEM, Armstrong AW, Gudjonsson JE, Barker JNWN. Psoriasis. Lancet. 2021;397(10281):1301-1315.
9. Singh SD, & Rao GP. Stevia: The herbal sugar of the 21st century. Sugar Tech. 2005;7(1):17-24. https://doi.org/10.1007/BF02942413
10. Tiwari S. Pharmacognostic and phytochemical evaluation of Stevia rebaudiana Bertoni leaves. Asian Journal of Pharmaceutical and Clinical Research. 2010;3(3):224-226.
11. Wagner H, & Bladt S. Plant drug analysis: A thin layer chromatography atlas (2nd ed.). Springer.1996.
12. Yadav AK, Singh S, Dhyani D, & Ahuja PS. A review on the improvement of stevia [Stevia rebaudiana (Bertoni)]. Canadian Journal of Plant Science. 2011;91(1):1-27. https://doi.org/10.4141/cjps10086
13. Chaturvedi P, & George S. HPTLC analysis of steviol glycosides in Stevia rebaudiana. Journal of Planar Chromatography. 2013;26(3):226-230.
14. JECFA (Joint FAO/WHO Expert Committee on Food Additives). Safety evaluation of steviol glycosides. WHO Food Additives Series, 2010;60: 183-218.
15. Kim IS, Yang M, Lee OH, & Kang SN. The antioxidant activity and the bioactive compound content of Stevia rebaudiana water extracts. LWT-Food Science and Technology. 2011;44(5):1328-1332.
16. Mizutani K, & Tanaka O. Use of Stevia rebaudiana sweeteners in Japan. In Stevia: The genus Stevia. CRC Press. 2002;178-195.
17. Prakash I, DuBois GE, Clos JF, Wilkens KL, & Fosdick LE. Development of rebiana, a natural, non-caloric sweetener. Food and Chemical Toxicology. 2008;46(7):S75-S82.
18. Shivanna N, Naika M, Khanum F, & Kaul VK. Antioxidant, anti-diabetic, and renal protective properties of Stevia rebaudiana. Journal of Diabetes and Its Complications. 2013;27(2):103-113.
19. Tadhani MB, Patel VH, & Subhash R. In vitro antioxidant activities of Stevia rebaudiana leaves and callus. Journal of Food Composition and Analysis. 2007;20(3-4):323-329.
20. Wolwer-Rieck U. The leaves of Stevia rebaudiana (Bertoni), their constituents, and the analyses thereof: A review. Journal of Agricultural and Food Chemistry. 2012;60(4):886-895.
21. Yoda SK, Ferreira ME, & Pessoa OD, Supercritical fluid extraction from Stevia rebaudiana Bertoni using CO₂ and CO₂+water: Extraction kinetics and identification of extracted components. Journal of Food Engineering. 2003;57(2):125-134.
22. Zhang Q, Yang H, Li Y, Liu H, & Jia X. Toxicological evaluation of ethanolic extract from Stevia rebaudiana Bertoni leaves: Genotoxicity and subchronic oral toxicity. Regulatory Toxicology and Pharmacology. 2017;86:253-259.
Statistics
63 Views | 43 Downloads
How to Cite
Jay Prakash Singh, and Shikha Sharma. “PHYTOCHEMICAL SCREENING, PHARMACOGNOSTICS CH- ARACTERIZATION, AND CHROMATOGRAPHIC ANALYSIS (TLC/HPTLC) OF STEVIA REBAUDIANA BERTONI LEAF EXTRACT”. Current Research in Pharmaceutical Sciences, Vol. 15, no. 2, Aug. 2025, pp. 51-57, doi:10.24092/CRPS.2025.150204.
Section
Research Articles