A novel extraction of trichosanthin from Trichosanthes kirilowii roots using three-phase partitioning and its in vitro anticancer activity.
BACKGROUND
Three-phase partitioning (TPP), a unique technique which has been explored for protein separation, was used for extraction of trichosanthin (TCS).


OBJECTIVE
TPP was used to optimize the TCS extraction and to determine its anticancer activity.


METHODS
The process consists of the simultaneous addition of t-butanol and ammonium sulfate to the aqueous slurry of Trichosanthes kirilowii Maxim (Cucurbitaceae) root powder. The extraction of TCS was optimized with respect to the concentration of ammonium sulfate loading, the ratio of t-butanol to slurry, extraction time and pH. The anticancer activity was performed using 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) assay in vitro.


RESULTS
The extraction time with this technique is lower in comparison to conventional solvent extraction. The optimized protocol resulted in maximum recovery of 98.68% (w/w) protein within 1 h. The in vitro cytotoxic activity of the TCS was evaluated against HepG2 and WRL 68 cancer cell line and results showed that TCS possesses quite highly significant anticancer activity having IC50 values of 10.38 and 15.45 μmol/l, respectively, comparable to standard drugs.


CONCLUSIONS
This framework is utilized as a basis for optimization for protein separation using TPP technique which is economical and eco-friendly.
Drug Development Diagnostics & Biotechnology Division, Indian Institute of Chemical Biology , Kolkata , India.
Arijit
A
Mondal
D000972:Antineoplastic Agents, Phytogenic / Q000302:isolation & purification* / Q000494:pharmacology; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D004305:Dose-Response Relationship, Drug; D006367:HeLa Cells; D056945:Hep G2 Cells; D006801:Humans; D010936:Plant Extracts / Q000302:isolation & purification* / Q000494:pharmacology; D018517:Plant Roots; D029778:Trichosanthes; D015978:Trichosanthin / Q000302:isolation & purification* / Q000494:pharmacology
D000972:Antineoplastic Agents, Phytogenic; D010936:Plant Extracts; D015978:Trichosanthin
HepG2; MTT assay; SDS-PAGE; WRL 68