Black-Bone Silky Fowl (Gallus gallus domesticus Brisson) is both consumed as a healthy food and used particularly as a kind of traditional Chinese medicine. Carnosine in the muscles of Black-Bone Silky Fowl was identified by HPLC–MS/PAD. The carnosine content in the meat of Black-Bone Silky Fowl was determined by HPLC and compared with that contained in the meat of White Plymouth Rock, which were bred under the same condition. The results showed that the contents of carnosine in the mixed meat, breast meat and thigh meat of Black-Bone Silky Fowl were all remarkably higher than that in the White Plymouth Rock. These findings indicate that Black-Bone Silky Fowl would be a better chicken breed for carnosine supplement.