Abstract:A novel determination method was developed for the phosphopeptide of goat milk which based on MALDI-TOF MS technology.Primarily，commercial TiO2was recommended for the process of phosphopeptide enrichment，and the optimized resultofenrichmentprocess suggests thatthe content ofphosphopeptide reached a maximum by using acetonitrile∶water（50∶50，volume ratio）+1%trifluoroacetic acid as the incubation buffer while using 15%ammonia as eluent.Subsequently，the effect of remove fat from milk for the detection resultwas discussed，and the resultItconfirmed thatitis necessary to remove fat from milk before make a more complete identification ofthe phosphopeptides in milk samples.MALDI-TOF MS was used to determine the enriched phosphopeptide of goat milk.Through fingerprints analysis and comparison with the Mascot database，eight phosphopeptides have been identified in goatmilk.The present study provides the theory base for the establishmentofthe dairy phosphopeptide fingerprintand contributes for rapid identification ofadulterated milk.