A detection chip is provided. The detection chip includes a substrate, an active reagent, a hydrophilic droplet and a lipophilic substance. The substrate includes a first containing slot, wherein the first containing slot includes a first space and a second space adjacent to each other. The active reagent is disposed in the first space of the first containing slot. The hydrophilic droplet is disposed in the second space of the first containing slot. The lipophilic substance is disposed in the first containing slot, wherein the lipophilic substance is immiscible to the active reagent and the hydrophilic droplet, and separates the active reagent from the hydrophilic droplet.