I am trying to separate my active compound through HPLC. When I injected my samples, I get more than 20 peaks and I was collected all peaks into 10 different fractions. After doing bioassay, I have shown that all fractions was shown activity i.e means all fractions contained active compound. Is it possible?I dont understand. Why all fractions was shown activity--that means my compound not separated or others reasons. I am also surprised to see my bioassay result. But is it possible about 20 compounds have similar activity? I dont know. Actually it is a growth promoting compound which promote the growth of uncultured bacteria in laboratory conditions and its origin is also another bacteria. When I used equivalent concentration, all factions did not show similar actiivity but when I used 10 fold concentrations, all fractions shown similar activity. There is a possibility of concentrations effect but it is true, all factions have shown similar activity.
Please give me some suggestions