Abstract: In this study,thermosensitive and repairable molecularly imprinted solid-phase microextraction fibers were synthesized using spiramycin as template molecule,methacrylic acid and N-isopropylacrylamide as functional monomers,ethylene glycol dimethacrylate as crosslinking agent,and silanized quartz capillary as carrier. The prepared molecularly imprinted solid-phase microextraction fibers were characterized by scanning electron microscope and nitrogen adsorption/desorption,and various parameters affecting the extraction efficiency were optimized. Due to high selectivity and sensitivity of the fibers for macrolide antibiotics,the quantitative analysis of four macrolide antibiotics in food matrix,spiramycin,tilmicosin,tylosin,and josamycin,was peroformed in combination with high performance liquid chromatography. In the range of 0.5 to 50 μg/mL,the chromatographic peak area showed a good linear relationship with the concentration. The spike recoveries of the samples at three different addition levels were between 81.8% and 119.1%;the inter-day precisions were less than 13.8% (n=6),and the intra-day precisions were less than 15.5% (n=3).