A hollow sphere from amphiphilic chitosan derivatives and a method of preparing an amphiphilic chitosan derivative complex for medical use are disclosed, and the hollow sphere from amphiphilic chitosan derivatives comprises: chitosan derivatives represented by the following formula (I), which self-assemble and form a hollow sphere in a solventwherein, each R1 is independently hydrogen, C1˜C4 alkyl, C1˜C6 carboxyl, sulfate group, or phosphate group, each R2 is independently hydrogen, C1˜C12 alkyl, C1˜C6 carboxyl, or C2˜C12 acyl group, and m is an integer of 100-2000.