There will be two strategies to reveal the next story. First is to identify the major constituents of the optimized antioxidant contents and further purify them for the antioxidant assessments, which will then clarify which components dominate the activity or there're synergetic effects. Other strategy will be to comprehensively illustrate the constituents inside the optimized antioxidant extracts using MS/MS metabolic approach, such as GNPS.

Let's say that the compound have been identified as orientin and vitecin via HPLC analysis.. should the extract be carry out straight away using the crude extract containing those optimized antiox compounds for LC50 analysis against specific cancerous cell lines and later the synergistic mechanism at the molecular level? My attention is to proof the efficacy of those compounds against cancer cell? Any recommendation of looking up into that molecular level?

I would suggest to go on DPPH activity. I believed there're other constituents possess antioxidant effect rather than orientin and vitecin, or you can convince reviewers that these two are the only and major active compounds in the crude extract of Clinachanthus nutans. Therefore, to comprehensively clarify the relationships between activity and constituents will be necessary.

In fact that, there are many interesting stories to carry out the next directions. Here, I will recommend two typical outstanding articles of each direction as the available link:

Experiment:

Article Antioxidant Properties of Crocin from Gardenia jasminoides E...

Theory:

Article Radical Scavenging Activity of Natural Anthraquinones: a The...

Best regards.