Phytochemical Methods

In this subject the students familiarize with the special isolation and analytical procedures, which are involved in the work with plant material for the effective isolation of compounds of different types and for determination of content of secondary metabolites by the modern analytical methods.

In the practical classes the students familiarize with the procedure of processing of row material which leads to the identification of compounds.

The education is directed to creative approach during the isolation of compounds and phytochemical analyses. The subject is intended for applicants who are interested in the diploma thesis with phytochemical theme.



General part (includes explication of use of optimal and special materials for phytochemical work):

  • Laboratory material
    • material on the bases of silicon dioxide, metal material, rubber, plastics, paper and other materials
    • laboratory apparatuses
  • fragmentation, netting, sampling, storage and documentation of samples
  • separation methods
    • extraction, decantation and filtration
    • chromatography
    • crystallization
  • evaluation of isolated compounds
    • identification of structure on the bases of physical and physiochemical methods (methods VIS, UV, IR, OR, MS, NMR)
  • procedures of analytical phytochemistry on the selected examples
    • use of methods of HPLC
    • use of methods of GC

Special part (includes model explication of the most frequent structure types of secondary metabolites)

  • simple phenols (except flavonoids)and phenolic acids (aglycones, glycosides, acids derivatives)
  • coumarins
  • lignans
  • diarylheptanoids and arylalkanones
  • stilbenoids
  • xanthones and styrylpyrones
  • flavonoids, isoflavonoids, neoflavonoids and flavanoids
  • anthocyanins
  • tannins
  • polyketides
  • naphtoquinones, anthraquinones, naphtodianthrones, quinones
  • orcinols and floroglucinols
  • terpnoids (monoterpenes, sesquiterpenes and lactones, diterpenes, triterpenes, carotenoids)
  • different types of steroids, cardiotonic glycosides
  • different types of alkaloids

Practical classes

  • preparation of basic equipment made of laboratory glass (capillary tubes for spotting of solution on TLC plates, adjusting capillary tubes, Pasteur’s capillary tubes, filter balloons, balloons for Svoboda’s crystallization technique, and others)
  • preparation and fractionation of extracts
  • thickening of extracts, distillation of volatile constituents
  • column chromatography
  • thin layer chromatography (analytical, preparative)
  • crystallization
  • basic physical characteristic (melting point, boiling point, optical rotation)


  • Mayo, D. W., Pike, R. M., Butcher, S. S., Trumper, P. K.: Microscale Techniques for the Organic Laboratory. John Wiley and Sons Inc., New York 1991, 285 pp., ISBN 0471-62192-7 (paperback)
  • Ikan, R.: Natural Products. A Laboratory Guide. 2nd Edition, Academic Press, Inc., Sant Diego 1991, 360 pp. ISBN 0-12-370551-7.
  • Canell, R. J. P.: Natural Products Isolation. Humana Press, Totowa, New Jersey 1998, 473 pp. ISBN 0-89603-362-7

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