Optimization Of Bioactive Compotents Extraction From Curcumae Rhizoma For Anti-proliferative Activity

Herbal medicine sharing characteristics of high pharmacological activities,low toxicity and rare complications are invaluable drug resources.Active ingredients in the Chinese medicine are the basis of its pharmacological actions.Different extraction methods achieved different effective components,then the biological activities of the extracts may be different.Curcumae Rhizoma,a member of the genus Curcuma,contains a variety of bioactive components.While,there is no study on optimizing extraction of three or more components from Curcumae Rhizoma.Taking Curcumae Rhizoma as the test material,this work optimizated the ultrasonic-assisted extraction（UAE）conditions of extraction of several compounds from Curcumae Rhizoma using response surface methodology（RSM）.The main results include:（1）A novel dual wavelength gradient elution high performance liquid chromatography（HPLC）coupled with UAE method was developed for simultaneously quantitative determination of sesquiterpenoids and curcuminoids,including curdione,furanodienone,curcumol,germacrone,furanodiene,β-elemene,curcumin,demethoxycurcumin and bisdemethoxycurcumin,that are the main bioactive ingredients in Curcumae Rhizoma.The HPLC method proposed is simple,accurate,sensitive and reproducible,and provides a new analytical approach to effectively evaluate and monitor the quality of Curcumae Rhizome in clinical use.（2）The RSM combining central composite design was used to investigate the effects of processing parameters on the dependent variables:extraction contents of curdione,furanodienone,curcumol and germacrone from Curcumae Rhizoma.Mathematical models were established by implementing RSM,and desirability function（DF）was employed to combine the four responses to one.The results of RSM and DF revealed that the highest desirability 0.971 was obtained with 8 mL·g^-1 for liquid-solid ratio,70%ethanol concentration and 20 min of ultrasonic time.The surface structures of the sonicated Curcumae Rhizoma materials were fluffy and irregular.（3）The anti-proliferative activities of the extract of Curcumae Rhizoma,with these four compounds alone or in combination,against RKO and HT-29 cells,were determined using a CCK-8 test.The extract of Curcumae Rhizoma obtained under the optimal UAE conditions exhibited significant anti-proliferative effects on RKO and HT-29 cells for 48h in vitro.Furanodienone displayed the highest inhibitory effect on the proliferation of RKO and HT-29 cells among the four compounds.In summary,this study successfully optimizated the UAE conditions for extracting of curdione,furanodienone,curcumol and germacrone from Curcumae Rhizoma employing RSM.Besides,the Curcumae Rhizoma extract exhibited significant anti-proliferative effects on RKO and HT-29cells in vitro,establishing foundation for further rational development and utilization of Curcumae Rhizoma.