A high performance liquid chromatography assay method for spiramycin in fish muscle was developed. The developed method was evaluated and validated by monitoring spiramycin in olive flounder (Paralichthys olivaceus), black rock fish (Sebastes schlegeli) and in live conger eel (Anguilla japonica) in fish farms and distribution centers. Using the developed method, the recovery rate was up to 82.4-88.8%, which was higher than that of conventional methods (77.6-87.1%). In particular, the proposed sample treatment protocol was suitable for use with fish samples to remove low molecular weight materials and pigments that could interfere an accurate analysis. The prepared stock solution was very stable, and it remained chemically stable for 5 weeks at $4^{circ}C$. The performance limit of the developed method for spiramycin acid in fish muscle was 0.05 ppm. One hundred thirty-four fish samples including olive flounder, black rock fish and live conger eel were analyzed to evaluate the overall efficiency of the modified method and to monitor the actual condition of spiramycin usage in fish farms. Olive flounder and black rock fish were collected from fish farms in coastal areas of Korea, and live conger eels were purchased from a fish market in the Busan area from September 2001 to March 2002. According to the overall performance of the developed method, it was considered suitable for the monitoring of spiramycin in fish muscle. The suggested method of analysis for spiramycin in fish muscle is registered in the Korean Official Methods of Food Analysis and is currently tieing utilized for fishery products inspection by the Korea Food and Drug Administration and the National Fisheries Products Quality Inspection Service.