The intrinsic d-amino acid profile of mouse macrophages extracted from the peritoneal cavity was analyzed using high performance liquid chromatography. Six d-amino acids (d-Asp, d-Ser, d-Ala, d-Leu, d-Gln and d-Lys) were detected in cell lysates of mouse macrophages. The content and the d/d + l ratio differed depending on the type of d-amino acid and were approximately 3.5–22 nmol/g cells, and approximately 1–20%, respectively. The d-amino acid composition of RAW 264.7 cells, which is a model macrophage cell line, was similar to that of the mouse macrophage. These results suggest that macrophages and RAW 264.7 cells with macrophage-like functions have a similar d-amino acid profile.