Well-defined poly (2-(dimethylamino) ethyl methacrylate) (PDMAEMA) brushes were successfully prepared on the monolithic surface via two-step atom transfer radical polymerization (ATRP). The polymer brushes synthesized by the second-step ATRP were based on the active bromic groups resulting from poly (ethylene glycol dimethacrylate) (pEDMA) monolith which was prepared at room temperature by the first-step ATRP. Element analysis was used to monitor the grafting process at different reaction times. Each step of preparation was characterized by scanning electron microscope, infrared spectrum and mercury intrusion porosimetry. Employment of PDMAEMA grafted monolith as the stationary phase for chromatographic analysis of steroids demonstrated that the PDMAEMA brushes possessed both pH- and salt-responsive properties. Noticeably, it has been found that the chain length of PDMAEMA brushes could influence the retention behavior of steroids due to the controllability of ATRP, which proposed an interesting alternative to modulate retention in HPLC. This is the first application of PDMAEMA brushes grafted monolith by two-step ATRP method for constructing responsive surface in HPLC and it might exploit a new path for widening the monolith application in various fields. (C) 2011 Elsevier Ltd. All rights reserved.