Estabelecimento de cultura de células em suspensão e identificação de flavonóides em Cordia verbenacea DC.

Abstract

The aims of this study were to establish cell suspension cultures, as well as to extract, separate and identify flavonoids in Cordia verbenacea leaf extracts and cell suspensions. Cells of this species were subcultivated three times in MS culture medium supplemented with 2.32 µM kinetin + 10.74 µM NAA at 28-day intervals, showing five growth stages: lag, exponential, linear, deceleration and stationary phases. The highest growth rate (37%) occurred in the exponential phase between the fourth and the twelfth day and the lowest growth rate (3%), in the lag phase until the fourth day. For flavonoid identification, extracts were separated and purified by TLC and LC and the obtained compounds were subjected to Ultraviolet Spectroscopy, Infrared and Mass Spectrometry, and Hydrogen Nuclear Magnetic Resonance. After leaf and cell sample fractions were separated using the eluent acetic acid, the compounds 7,4'-dihydroxy-5'-carboxymethoxy isoflavone and 7,4'-dihydroxy-5'-methyl isoflavone were identified. Higher concentrations of these substances were obtained in in vitro-cultivated cells.