The invention provides a disposable separating chip module for a peripheral blood circulating tumor cell and an application method of the disposable separating chip module. The disposable separating chip module comprises a micro-fluidic chip, a blood sample storage tank, a PBS sample storage tank, a waste liquid storage tank, an enrichment recycling liquid outlet, a connecting guide pipe (1) of the micro-fluidic chip and the blood sample storage tank, a connecting guide pipe (2) of the micro-fluidic chip and the PBS sample storage tank, a connecting guide pipe (3) of the micro-fluidic chip and the waste liquid storage tank, and a connecting guide pipe (4) of the micro-fluidic chip and the enrichment recycling liquid outlet. The invention further provides the application method of the disposable separating chip module. The micro-fluidic chip adopts a dual-inlet and dual-outlet structure and has an erythrocyte removing efficiency of 100%. A clinical blood sample requires no pretreatment process of attenuation or erythrocyte lysis, so that plenty of time for separating operation can be saved, and the activity of the circulating tumor cell after separation and enrichment can be effectively guaranteed.