Abstract:

Javanese chili (Piper retrofractum Vahl) is a traditional medicinal plant originating from Indonesia and has many pharmacological activities, one of which is often used as a base for aphrodisiac herbal medicine. The population of P. retrofractum is limited, so it is necessary to design secondary metabolite production and propagation efforts using plant tissue culture techniques. The materials used in this study were explants of . retrofractum leaves that were induced in Murashige and Skoog (MS) media and the ratio of growth regulators 2.4-dichlorophenoxyacetis acid (2,4-D): Benzyl Amino Purine (BAP) 0.5: 0.5. The results showed that the callus of P. retrofractum was formed in the growth regulator 2.4D: BAP (0.5: 0.5). TLC and spectrophotometry identified the secondary metabolite content of callus. Secondary metabolite analysis using the thin layer chromatography (TLC) method using the mobile phase ethyl acetate: n-hexane (7: 3) showed a terpenoid compound indicated by purple spots on the visual appearance after spraying 10% spotting vanillin. Identification using infrared spectrophotometry shows functional groups -CH, C = O, C = C, -CH2, and CH3, characteristic of terpenoid compounds. Based on TLC data and spectrophotometry, callus P. retrofractum is thought to contain terpenoid compounds. This study's results are expected to be the basis for developing secondary metabolite production in P. etrofractum with cell suspension culture and P. retrofractum propagation by in vitro culture.

Keywords: P. retrofractum Vahl, Plant tissue culture, Callus, Identification of secondary metabolites