Abstract: Liquid chromatography combined with mass spectrometry has been widely used in proteomics research,especially the development of two-dimensional chromatography provides a more accurate technical method for the separation and analysis of complex biological samples.The hippocampus is an important brain region of the medial temporal lobe,mainly responsible for learning and memory.These functions are closely related to the hippocampal membrane protein which is difficult to separate and identify due to its hydrophobicity and low abundance.Hippocampal membrane protein was isolated and purified By the way of differential centrifugation.The RP-RPLC-MS/MS was used for the separation and analysis of hippocampal membrane protein.2502 proteins was identified,which were further found involved in many process related to learning and memory through the bioinformatics analysis,including the process of protein localization,synaptic plasticity,protein transport and vesicular transport.This study will help to reveal the relationship between hippocampal membrane protein and function.