CSSP or CP NPs (300 nmol) were incubated with 10, 1, or 0 mM DTT in 30 ml of PBS (pH 7.4) [30% ethanol (v/v)] at 37°C (n = 3). The released CPT and PR104A were simultaneously determined by HPLC. In addition, the molecular weight change of prodrugs was measured by Agilent 1290-6545 Q-TOF (Agilent, USA) to investigate the release mechanism after CSSP NPs were incubated with 1 mM DTT for approximately 2 hours.

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