Background: Various methods of isolating compounds from plants have been described
previously, which include conventional or modern techniques. A preparative-HPLC (prep-HPLC)
system has become one of the most convenient methods, with high purity compound as well as consumes
less purification time.
Objective: This study is intended to purify compounds from E. pulchrum twig extract using prep-
HPLC technique and to test all compounds in several biological activities.
Methods: Prior to purification using prep-HPLC, the twig extract was injected onto HPLC to develop
the method through its chromatograms. The established method from HPLC was used to separate
the constituents using prep-HPLC. Purified compounds were elucidated through NMR and MS
methods as well as through comparison with previously reported data. Three different biological activities
were then conducted on the compounds, including cytotoxicity, DPPH, FRAP, and disc diffusion
Results: Cinnamic acid (1) and two aporphine alkaloids (liridine (2) and lysicamine (3)) have been
successfully purified and identified. These compounds were first isolated from Enicosanthellum
pulchrum using prep-HPLC. Cytotoxic activity revealed that liridine (2) showed strong inhibition
against WEHI-3B leukaemic cells of 8.7 µM after 24 h of treatment. In contrast, cinnamic acid (1)
and lysicamine (3) exhibited strong inhibitions in antibacterial activity against Staphylococcus
aureus, S. epidermidis, Bacillus cereus, Pasteurella multocida and B. subtilis with more than 15 mm
of inhibition zone.
Conclusion: These phytochemical findings exhibit three isolated compounds from twig extract of E.
pulchrum with diverse biological potential to be developed as new agents.