Chromatographia

, Volume 75, Issue 23–24, pp 1445–1449 | Cite as

An Improved Method to Resolve Plant Saponins and Sugars by TLC

Short Communication

Abstract

Thin-layer chromatography (TLC) plays an important role in the initial selection of mutants having a unique seed saponin composition from the germplasm collections of the subgenus Soja. In the conventional TLC procedure, the dehydrated free sugars are retained just below the major saponins and interrupt the identification of some minor saponin constituents. To resolve this problem, we developed an efficient and reliable method to move sugars from the saponin area on TLC. A developing chamber was saturated with the lower phase of chloroform:methanol:water (65:35:10, v/v) for 2 h and the TLC plates were developed in it for 50 min. Plates were then dried at 100 °C for 10 min to evaporate the excess mobile phase and developed again with 10 % H2SO4 for 15 min. While sulfuric acid migrates over the surface of SiO2, sugar molecules are dehydrated and hydrophilic interactions between free sugars and SiO2 are strongly reduced. Thus, the positions of dehydrated sugars were shifted to above the saponin area on the TLC plate. This resulted in easy recognition of the saponin composition without any discrimination. This amended protocol would be applicable to all TLC analyses in which the target components should be separate from the interrupting sugar molecules.

Keywords

Thin-layer chromatography Seed saponin composition Hydrophilic interaction Sugar Subgenus Soja 

Supplementary material

10337_2012_2340_MOESM1_ESM.docx (3.2 mb)
Supplementary material 1 (DOCX 3265 kb)

References

  1. 1.
    Price KR, Johnson IT, Fenwick GR (1987) Crit Rev Food Sci Nutr 26:27–135CrossRefGoogle Scholar
  2. 2.
    Shiraiwa M, Kudo S, Shimoyamada M, Harada K, Okubo K (1991) Agric Biol Chem 55(2):315–322CrossRefGoogle Scholar
  3. 3.
    Shiraiwa M, Harada K, Okubo K (1991) Agric Biol Chem 55(4):911–917CrossRefGoogle Scholar
  4. 4.
    Tsukamoto C, Kikuchi A, Harada K, Kitamura K, Okubo K (1993) Phytochemistry 34:1351–1356CrossRefGoogle Scholar
  5. 5.
    Kudou S, Tonomura M, Tsukamato C, Uchida T, Sakabe T, Tamura N, Okubo K (1993) Biosci Biotechnol Biochem 57:546–550CrossRefGoogle Scholar
  6. 6.
    Tsukamoto C, Yoshiki Y (2006) Soy saponin. In: Sugano M (ed) Soy in health and disease prevention. CRC Press, Taylor and Francis group, New York, pp 155–172. ISBN 0-8493-3595-7Google Scholar
  7. 7.
    Okubo K, Iijima M, Kobayashi Y, Yoshikoshi M, Uchida T, Kudou S (1992) Biosci Biotech Biochem 56:99–103CrossRefGoogle Scholar
  8. 8.
    Shibuya M, Nishimura K, Yasuyama N, Ebizuka Y (2010) FEBS Lett 584:2258–2264CrossRefGoogle Scholar
  9. 9.
    Chung G, Singh RJ (2008) Crit Rev Plant Sci 27:295–341CrossRefGoogle Scholar
  10. 10.
    Poole CF (1999) J Chromatogr A 856:399–427CrossRefGoogle Scholar
  11. 11.
    Selvam P, Tsukamoto C, Lee JD, Yang SH, Kim HS, Chung G (2011) Abstracts of the Korean Society of Crop Science, October 20–21, Danyang, Chungbuk, Korea. Korean J Crop Sci 56(2):17Google Scholar
  12. 12.
    Geiss F (1987) Dr. Alfred Hüthig Verlag, Heidelberg, GermanyGoogle Scholar
  13. 13.
    Shimoyamada M, Okubo K, Yoshikoshi M, Yoshiki Y, Watanabe K (1995) Food Sci Technol Int 1:112–114Google Scholar
  14. 14.
    Meeks EG (1979) Sch Sci Rev 61(215):281–283Google Scholar
  15. 15.
    Shiraiwa M, Yamauchi F, Harada K, Okubo K (1990) Agric Biol Chem 54:1347–1352CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2012

Authors and Affiliations

  • Panneerselvam Krishnamurthy
    • 1
  • Chigen Tsukamoto
    • 2
  • Seung Hwan Yang
    • 3
  • Jeong Dong Lee
    • 4
  • Gyuhwa Chung
    • 1
  1. 1.Department of BiotechnologyChonnam National UniversityYeosuKorea
  2. 2.Department of Applied Biological ChemistryIwate UniversityMoriokaJapan
  3. 3.Center for Nutraceutical and Pharmaceutical MaterialsMyongji UniversityGyeonggiKorea
  4. 4.School of Applied BiosciencesKyungpook National UniversityDaeguKorea

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