Investigation of phenolic compounds in the herb of hypericum elongatum by thin-layer chromatography

Introduction: The TLC method makes it possible to separate mixtures of medicinal substances, identify them, establish purity and carry out quantitative determination of components. TLC has the advantage of allowing the separation of complex drug mixtures into individual components that are similar in chemical structure and properties [1].

Purpose of the research. Definition (control) of the individuality of the substance; determining the identity of the test sample to the control; determining the specificity of the qualitative composition of complex mixtures.


Materials and methods. The object of the research was the herb of the Hypericum elongatum, collected and harvested in May-June 2017 in the Baydibek and Tolebi districts of the South Kazakhstan region during the flowering period of the plant.

For the analysis we used the method of thin-layer chromatography.

Results and discussion. In order to research the flavonoid compounds, we used alcohol extract (water-alcohol extract). In order to establish the identity of the alleged biologically active substances, flavonoid compounds, we used the method of thin-layer chromatography. Thin layer chromatography was carried out on plates "Sorbfil PTLC-АV-А-UV" (Russia) size 10x15. Microsprints (MS, Russia) and 2 mkl microcapillaries (Russia) were used to apply samples to the plates. The aqueous-alcoholic fraction was applied to the plate of a volume of 10 mkl, and solutions of standard samples in a volume of 5mkl were applied nearby. Further, chromatography was carried out in an upward manner. Then we showed spots in chromatograms using a 10% alcoholic solution of aluminum chloride and ammonia vapors, followed by UV irradiation. Chromatograms were viewed in UV light at a wavelength of 254 nm before and after treatment with chromogenic reagents.The optimal separation of the components of the water-alcohol mixture was observed in the solvent system n-butanol-acetic acid-water (6: 2: 3) [2].

After the development of spots, the chromatogram was identified by the Rf values of the studied substances and standard samples: rutin Rf ~ 0.51; Hyperoside Rf ~ 0.35; Quercetin Rf ~ 0.43; = 254 nm). Fluorescent brown or dark spots were detected in UV light ( = 254 nm), yellow-green, bright yellow after treatment with a 10% alcohol solution of aluminum chloride, detected also by UV-light ( = 254 nm).

Conclusion. A qualitative analysis of the raw materials confirmed the presence of phenolic compounds in the aerial part of the Hypericum elongatum L.. We have proven the presence of phenolic compounds such as flavonoids.



  1. 1. Harry W. Lewis & Christopher J. Moody (13 Jun 1989). Experimental Organic Chemistry: Principles and Practice (Illustrated ed.). WileyBlackwell. pp. 159–173. ISBN 978-0-632-02017-1.
  2. 2. Бандюкова В.А. Фенолокислоты растения, их эфиры и гликозиды // Химия природных соединений. №3. - 2003. - С. 263-273 [Bandyukov V.A. Phenolic acids of plants, their esters and glycosides // Chemistry of natural compounds. Number 3. - 2003. - p. 263-273].
Year: 2018
City: Shymkent
Category: Medicine