Isolation of the major compounds from the aerial parts of Chresta martii by high speed counter current chromatography (HSCCC)

Chresta martii (DC.) H. Rob. (Asteraceae) is a plant found in the Xingó region (caatinga ecosystem) in North-eastern in Brazil, and is recognized by the local population as a traditional herb used to treat gastric diseases. Previous studies demonstrate significant in vivo protective effects of C. martii extract against indomethacin-induced gastric lesions in mice [1]. In order to isolate the active compounds and avoid any possible adsorption that can occur on silica gel columns, High Speed Counter Current Chromatography (HSCCC) was used for the first fractionation step. The coefficient of partition (Kp) of each compound was first calculated using the peak area obtained by the HPLC-UV-ELSD analysis. The best conditions were obtained with using the quaternary system of heptane/EtOAc/MeOH/H₂O (8.3:41.7:8.3:41.7, v/v/v/v). The separation of the crude extract (3 g) was achieved in one step. Fractions and pure compounds were monitored by HPLC-UV-ELSD. Using these approach sesquiterpene lactones and flavonoids were isolated in one step and some minor compounds purified on a second step by semi-prep HPLC. Their structures were elucidated by spectroscopic and chemical methods and the in vivo bioassay on the pure constituents are underway.

References:

[1] Silva AA, Bezerra MM, Chaves HV, Pereira KM, Aguiar JA, Pinto VP, Abbet C, Simões-Pires CA, Franco ES, Henriques AT, Hostettmann K, Maia MB. Protective effect of Chresta martii extract against indomethacin-induced gastric lesions in mice. Nat Med 2013; 67:143 – 151.