Fractionating the Plant Extract in Mini Volumes to Purify the Potential Antivirals from Terminalia Chebula

Innokentii S. Solovarov, Maxim A. Khasnatinov, Tatyana A. Shishlyannikova, Natalie A. Lyapunova, Galina A. Danchinova

International Journal of Biomedicine. 2019;9(1):57-60.
DOI: 10.21103/Article9(1)_OA11
Originally published March 15, 2019


Background: The preliminary experiments indicated that a 30% aqueous extract of seeds of the Terminalia chebula has a significant neutralizing activity against tick-borne encephalitis virus (TBEV). In this study, we developed the most gentle approach to purify the antiviral substance.
Methods and Results: The extract of T. chebula seeds was fractionated using gel filtration in Sephadex G-200 and G-50. The fractions were tested in vitro for the presence of a direct antiviral action against TBEV and profiled using liquid chromatography-mass spectrometry (LC-MS). Antiviral activity was detected in eight fractions from Sephadex G-200 and in three fractions from Sephadex G-50 gel filtration. The chromatogram of the most active Sephadex G-50 fraction exhibited three base peaks with a retention time of 3, 5 and 25.5 minutes. The MS of individual peaks revealed seven compounds with mass-to-charge ratios 114.03; 279.16; 290.91; 301.15; 579.3; 354.03, and 414.09 that were present at relatively high concentrations.
Conclusion: In spite of a small sample, it was possible to detect antiviral activity in the fractionated plant extract using convenient virological methods, as well as to separate the components of the single virus-neutralizing fraction using the LC-MS approach. The purity of resulting preparation needs to be improved.

flavivirus • novel antiviral activity • chromatography • mass spectrometry
  1. Burke DS, Monath TP. Flaviviruses. In: Knipe DM, Howley PM, editors. Fields Virology. Lippincott-Williams & Wilkins, Philadelphia, PA; 2001:1043–1125.
  2. Lindenbah BD, Rice CM. Molecular biology of flaviviruses. Adv Virus Res. 2003;59:23–61.
  3. Prevention of tick-borne viral encephalitis. Sanitary rules SP3.1.3.2352-08. Moscow: Federal Service for Supervision of Consumer Rights Protection and Human Welfare; 2008. [In Russian].
  4. Weiss HJ, Aledort LM. Impaired platelet/connective-tissue reaction in man after aspirin ingestion. Lancet. 1967;2(7514):495-7.
  5. Lubbe A, Seibert I, Klimkait T, van der Kooy F. Ethnopharmacology in overdrive: the remarkable anti-HIV activity of Artemisia annua. J Ethnopharmacol. 2012;141(3):854-9. doi:10.1016/j.jep.2012.03.024.
  6. Cesar GZ, Alfonso MG, Marius MM, Elizabeth EM, Angel CB, Maira HR, et al. Inhibition of HIV-1 reverse transcriptase, toxicological and chemical profile of Calophyllum brasiliense extracts from Chiapas, Mexico. Fitoterapia. 2011;82(7):1027-34. doi: 10.1016/j.fitote.2011.06.006.
  7. Yu JS, Kim JH, Lee S, Jung K, Kim KH, Cho JY. Src/Syk-Targeted Anti-Inflammatory Actions of Triterpenoidal Saponins from Gac (Momordica cochinchinensis) Seeds. Am J Chin Med. 2017:45(3):459-473. doi: 10.1142/S0192415X17500288.
  8. Intisar A, Zhang L, Luo H, Kiazolu JB, Zhang R, Zhang W. Anticancer constituents and cytotoxic activity of methanol-water extract of Polygonum bistorta L. Afr J Tradit Complement Altern Med. 2012;1:53-9. eCollection 2012.
  9. Elisha IL, Botha FS, McGaw LJ  and Eloff JN.  The antibacterial activity of extracts of nine plant species with good activity against Escherichia coli against five other bacteria and cytotoxicity of extracts. BMC Complement Altern Med. 2017;17(1):133. Doi: 10.1186/s12906-017-1645-z.
  10. Webster D, Taschereau P, Lee TD, Jurgens T. Immunostimulant properties of Heracleum maximum Bartr. J. Ethnopharmacol. 2006;106(3):360–3. doi:10.1016/j.jep.2006.01.018.
  11. Yin SY, Yang NS, Lin TJ. Phytochemicals approach for developing cancer immunotherapeutics. Front Pharmacol. 2017;8:386. doi: 10.3389/fphar.2017.00386.
  12. Myagmar BE, Aniya Y. Free radical scavenging action of medicinal herbs from Mongolia. Phytomedicine. 2000; 7(3):221–9.
  13. Solovarov IS, Khasnatinov MА, Danchinova GА, Lyapunov АV, Bolotova NА, Manzarova EL, et al. Assessment of neutralizing properties of DNA-aptamers and extracts of medicinal herbs against the tick-borne encephalitis virus. Acta Biomedica Scientifica. 2017;2(1):84-88. [Article in Russian].
  14. Janiak M, Slavova-Kazakova A, Kancheva V, Amarowicz R.  Sephadex LH-20 column chromatography of the hydrolysed lignan macromolecule of flaxseed. Bulgarian Chemical Communications. 2014;46(3):640–644.
  15. Khasnatinov MА, Danchinova GА, Zlobin VI, Lyapunov АV, Аrbatskaya EV, Chaporgina EА, et al. [Tick-borne encephalitis virus in Mongolia]. Siberian Medical Journal (Irkutsk). 2012;111(4):9–12. [Article in Russian].
  16. Gould EA, Clegg JCS. Growth, titration and purification of togaviruses. In: Mahy BWJ, editor. Virology: A Practical Approach. IRL Press Ltd., Oxford; 1985:43-48.
  17. Oyuntsetseg N, Khasnatinov MA, Molor-Erdene P, Oyunbileg Ja, Liapunov AV, Danchinova GA, et al. Evaluation of direct antiviral activity of the DEVA-5 herb formulation and extracts of five Asian plants against influenza A virus H3N8. BMC Complement Altern Med. 2014;14:235. doi: 10.1186/1472-6882-14-235.

Download Article
Received December 4, 2018.
Accepted January 9, 2019.
©2019 International Medical Research and Development Corporation.