A new form of doxorubicin hydrochloride (DRH)-containing chitosan microspheres (CMs) was prepared by employing an expanding-loading-shrinking (E-L-S) process. One hundred mg of pre-formed CMs were soaked in absolute ethanol and then placed in reduced pressure (the expanding process). Ten mg of DRH (2 mg ml(-1)) were added into the expanded CMs (the loading process). Next the microspheres were freeze-dried (the shrinking process). As a result of this E-L-S process, 10% (w/w) DRH-containing CMs (DRH-CM) were made. During 7 days, 22.6% of the DRH was observed to be released on the in vitro drug release study. In addition, these new DRH-CMs could be used for transcatheter arterial chemoembolization (TACE) procedure in VX2 hepatic tumour models of rabbit and the anti-tumour effects of DRH-CMs were investigated. On the post-CT scan 7 days after the TACE, total infarctions of the VX2 tumour were observed in 5 rabbits among the 6 total rabbits.