To study the dissociation of ACC@DOX-CaSi-PAMAM-FA/mPEG complex in response to pH changes and their release profiles in solution, ACC@DOX-CaSi-PAMAM-FA/mPEG was dispersed into buffer solutions of different pH values (pH 7.4 and pH 5.5) at a concentration of 0.4 mg/ml. After predetermined periods of time, 0.2 ml of the incubation solution was extracted and centrifuged at 8000 rpm for 5 min to remove undissolved nanoparticles. The concentration of DOX in the supernatant was detected by a fluorescence spectrophotometer. The total amount of loaded DOX was determined similarly by dissolving 4 mg of ACC@DOX-CaSi-PAMAM-FA/mPEG in 10 ml of acidic buffer pH 5.5 and sonicated for 1 min. At this condition the ACC@DOX-CaSi-PAMAM-FA/mPEG will degrade completely and release all the encapsulated contents. The DOX fluorescence was measured by extracting 0.5 ml of the incubation.

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