Evaluation of anticancer bioactive compounds and cytotoxicity of citrus hystrix Dc. Callus extract post preservation

Previous studies have shown that kaffir lime leaf extract is toxic to cancer cells. To increase the bioactive compound's production for traditional cancer medicine, we induce kaffir lime callus in vitro. One strategy to continuously maintain the production of kaffir lime callus is by using callus preservation. Our preliminary study used two preservation methods of callus stored in 4°C with or without alginate encapsulation. However, low temperatures and sodium alginate can be stress factors for plants, affecting bioactive compounds' production and their anti-cancer ability. Therefore the objective of this study was to determine whether our preservation methods affect the character of the callus by evaluating the bioactive compounds of callus post preservation and their effect on the cytotoxicity against T47D and Vero cells. This study was conducted by inducing kaffir lime seeds to form callus. The generation 1 calluses were divided into control and preserved groups. Callus preservation was performed by stored callus in 4°C with or without alginate encapsulation for 21 days and then recultured for 14 days. The bioactive compounds in the callus extract are detected by GC-MS. Furthermore, cytotoxicity of callus against breast cancer (T47D) and non-cancer cell (Vero) are tested using the MTT method. The results showed that preservation in 4°C with and without encapsulation caused changes in bioactive compounds profile. The terpenoid compounds were detected post preservation are Squalene, Geranyl linalool, and Geranyl acetate. Other anti-cancer bioactive compounds such as Stearic acid, 1-Decanol, Octadecane, 1-Hexcosanol, Hexane, Dodecane, Tetracosane, and 2-Decenoid acid. However, control and post-preservation callus extract are not cytotoxic to both cancer and non-cancer cells. Although there was a slight difference in the type of bioactive compounds, those compounds might be synthesized at a minimal level thus they did not affect cytotoxicity. Our preservation method could well storage the callus thus it can be used to provide continuous supply callus stock for pharmaceutical purposes. Copyright © 2020 The Author(S).