A Novel Method To Separate,Identify And Research Biological Mechanism Of Capsicum Annuum L. And Tageteser Ecta L.

In order to research the active compounds from Capsicum annuum L. and the mechanism of active compounds from Tageteser ecta L. The preparative HPLC was used to separate and purify active compounds from Capsicum annuum L. The crude sample of Capsicum annuum L. was prepared on Silica column for the first dimensional separation. Then the sub-fractions were separated on Amide column for the second dimensional preparation. It showed that Silica column and Amide column had good orthogonality when the non-aqueous solutions were taken as mobile phases. As a result, two compounds with the purity of more than 97% were obtained.At the same time, the compounds from Capsicum annuum L. were screened the activities. The antioxidant and antitumor screening platform were established in our laboratory. A unique cellular antioxidant activity model using anti-oxidative response element to regulate the expression level of luciferase reporter gene was employed to evaluate the relative antioxidant activity of purified compounds with Tertiary Butyl Hydroquinone(TBHQ) as the positive control. A unit(LCAA) for the measurement of the antioxidant capacity was defined. The antioxidant capacity of 1.5 μg/m L TBHQ was defined as 1 LCAA. At last, the relative antioxidant abilities of Capsanthin and Capsorubin were 0.397 LCAA and 0.523 LCAA respectively. In the antitumor screening models, the iCELLigence system was used to evaluate the inhibitory on hepatocellular carcinoma cells and lung cancer cells. At last, the compounds from Capsicum annuum L. did not inhibit the cancer cells growth.What’s more, we also studied the mechanism of quercetagetin. Affinity chromatography was used to separate the target protein of quercetagetin. The metabolic pathways of proteins were analyzed by KEGG. At last, the protein K-Ras played an important role in the metabolic pathways of colorectal cancer and pancreatic cancer. The protein Myc was important in small cell lung cancer. This showed that quercetagetin might bind with K-Ras protein so that the anti-liver cancer effect of quercetagetin was good, which provided a reliable theory for the quercetagetin used as a new drug.