Modified Extraction method and Characterization of sweet compounds from Stevia Rebaudiana
Keywords:
Stevia rebaudiana, Asteraceae, Lyophilization, HPLC-PDA, Stevioside, Rebaudioside aAbstract
Stevia is a natural sweetener obtained from the leaves of Stevia rebaudiana (Bertoni), belonging to the
Asteraceae family. The objective of the present study was to develop a rapid and effective methodology
for the extraction of stevioside and rebaudioside A from Stevia leaves and to compare the yields with
different extraction techniques. Modified method of extraction was done with water finally the glycosides
were precipitated by methanol-water solvent (0.25:1.0 v/v). The solvent from the extract was removed
by lyophilization. Extracted product was analyzed by HPLC-PDA followed by mass-spectrometric (MS)
detections. The decolorized extract was separated on an NH2 column using PDA detection with excitation
at 210 nm. The mobile phase consisted of acetonitrile/water (80:20 v/v) with a flow rate of 1 ml/min.
Evaluative criteria for stevioside and rebaudioside yield in this new method was high. An improved
analytical method was developed which may be applied to quality control of stevioside content in dried
leaves of Stevia before processing.
References
derivatives and in vitro metabolites of steviol, the aglycone of the natural sweeteners, stevioside, rebaudioside A and rubusoside. Biomed. Environ. Mass Spectrom. 1998; 15(4): 211-222.
2. Medon PJ, Pezzuto JM, Havanec-Brown JM, Nanayakkara NP, Soejarto DD, Kamath SK. Safety assessment of some Stevia rebaudiana sweet principles. Fed Proc. 1982; 41: 1568-1982.
3. Geuns JMC. Molecules of Interest-Stevioside. Phytochemistry. 2003; 64(5): 913-921.
4. Hutapea AM, Toskulkao C, Buddhasukh D, Wilairat
P, Glinsukon T. Digestion of stevioside, a natural sweetener, by various digestive enzymes. J Clin Biochem
Nutr. 1997; 23(3): 177-186.
5. Jakinovich W, Moon C, Choi YH, Kinghorn AD. Evaluation of plant extracts for sweetness using the Mongolian
gerbil. J Nat Prod. 1990; 53(1): 190-195.
6. Kinghorn AD, Soejarto DD, Nanayakkara NPD, Compadre CM, Makapugay HC, Hovanec-Brown JM, Medon, PJ,
Kamath S, Nanayakkara. A phytochemical screening procedure for sweet ent-kaurene glycosides in the
genus Stevia. J. Nat. Prod. 1984; 47: 439- 444.
7. Haga T, Ise R, Kobayashi T. A method of purifying stevioside (English abstr.) Jap. Patent 51-131900. 1976.
8. Ogawa T. Decolorization and purificationof stevia sweet component (English abstr) Jap. Patent 55-111768.
1980.
9. Itagaki K, Ito T, Purification of Stevioside (English abstr) Jap. Patent 54-041898. 1979.
10. Uneshi H, Ise R, Kobayashi T. Purification of stevia sweetening agent (English abstr) Jap. Patent 54-
030199. 1977.
11. Matsushita K, Kitahara T. Separation of stevioside and rebaudiosideA by crystallization (English abstr.) Jap.
Patent 56-121454. 1981.
12. Phillips KC. Stevia: steps in developing a new sweetner, T.H.Grenby ed., Developments in sweeteners. Elsevier
Applied Science, London. 1989; 3: 41-43. .
13. Tan S, Ueki H. Methods of extracting and separating sweet substances of Stevia rebaudiana Bertoni, ( English abstr.) Jap. Patent 06-007108. 1994.
14. Tanaka O. Steviol- glycosides: new natural sweetners. Trends Anal.Chem.1982; 1: 246-248.
15. Fullas F, Kim J, Compadre CM, Kinghorn AD. Separation of natural product sweetening agents using over
pressured layer chromatography. J. Chromatogr., 1989; 464: 213-219.
16. Mizukami H, Shilba K, Ohashi H. Enzymatic determination of stevioside in Stevia rebaudiana. Phytochemistry 1982; 21: 1927-1930.
17. Liu J, Li SFY. Separation and determination of Stevia sweeteners by capillary electrophoresis and high
performance liquid chromatography. J. Liq. Chromatogr., 1995; 18: 1703-1719.
18. Shi R, Xu M, Shi Z, Fan Y, Guo X, Liu Y, Wang C, He B. Synthesis of bifunctional polymeric adsorbent and its application in purification of stevia glycosides. Reactive and Functional Polymers 2002; 50(2): 107-116.
19. Crammer B, Ikan R. Sweet glycosides from the stevia plant. Chem Br 1986; 22(10):915-917.
20. Wheeler A, Boileau AC, Winkler PC, Compton SC, Prakash, J., Jiang, X. Pharmacokinetics of rebaudioside
A and stevioside after single oral doses in healthy men. Food Chem Toxicol 2008; 46: S54-S60.
21. Pasquel A, Meireles MAA, Marques MOM, Petenate A J. Extraction of stevia glycosides with CO2+water, CO2+ethanol, and CO2+water+ethanol. Braz J Chem Eng 2000; 17(3): 271-282.
22. Pol J, Hohnová B, Hyötyläinen T. Characterisation of Stevia Rebaudiana by comprehensive twodimensional liquid chromatography time-of-flight mass spectrometry. Journal of Chromatography A 2007; 1150(1-2): 85-92.
Downloads
Published
How to Cite
Issue
Section
License
We, the undersigned, give an undertaking to the following effect with regard to our article entitled
“_______________________________________________________________________________________________________________________________________________________________________________
________________________________________________________________________________” submitted for publication in (Journal title)________________________________________________ _______________________________________________________Vol.________, Year _________:-
1. The article mentioned above has not been published or submitted to or accepted for publication in any form, in any other journal.
2. We also vouchsafe that the authorship of this article will not be contested by anyone whose name(s) is/are not listed by us here.
3. I/We declare that I/We contributed significantly towards the research study i.e., (a) conception, design and/or analysis and interpretation of data and to (b) drafting the article or revising it critically for important intellectual content and on (c) final approval of the version to be published.
4. I/We hereby acknowledge ADRs conflict of interest policy requirement to scrupulously avoid direct and indirect conflicts of interest and, accordingly, hereby agree to promptly inform the editor or editor's designee of any business, commercial, or other proprietary support, relationships, or interests that I/We may have which relate directly or indirectly to the subject of the work.
5. I/We also agree to the authorship of the article in the following sequence:-
Authors' Names (in sequence) Signature of Authors
1. _____________________________________ _____________________________________
2. _____________________________________ _____________________________________
3. _____________________________________ _____________________________________
4. _____________________________________ _____________________________________
5. _____________________________________ _____________________________________
6. _____________________________________ _____________________________________
7. _____________________________________ _____________________________________
8. _____________________________________ _____________________________________
Important
(I). All the authors are required to sign independently in this form in the sequence given above. In case an author has left the institution/ country and whose whereabouts are not known, the senior author may sign on his/ her behalf taking the responsibility.
(ii). No addition/ deletion/ or any change in the sequence of the authorship will be permissible at a later stage, without valid reasons and permission of the Editor.
(iii). If the authorship is contested at any stage, the article will be either returned or will not be
processed for publication till the issue is solved.