Abstract: Homocysteine (HCY) molecularly imprinted polymer microspheres were synthesized by suspension polymerization using homocysteine as template, acrylamide as functional monomer, ethylene glycol two methyl acrylate (EGDMA) as crosslinking agent, polyvinyl alcohol (PVA) as dispersant, azobisisobutyronitrile (AIBN) as initiator and toluene as diluent in water system of 55℃ at the speed of 800 r·min^-1. It was found that the prepared particles were uniform in size, the optimum adsorption time was 40 min, the maximum apparent adsorption capacity was 1.61 mg·g^-1, and the selectivity to other amino acids which structure were similar to HCY was very good. Linear relationship between values of absorbance and mass concentration of homocysteine was obtained within 24 mg·L^-1, with detection limit (3s) of 0.019 mg·L^-1. The proposed method was applied to the analysis of the sample of rabbit blood, giving values of recovery and RSDs (n=5) in the ranges of 98.4%-109% and 0.75%-3.5% respectively.