•The highest content in CRC tissue was found for C16:0-Cer (80.36% of total ceramide)•The highest content (and higher than in normal tissue) was demonstrated for C24:0-Cer and Sph•CRC tissue showed increased amounts of S1P, SPA, and C14:0-Cer•CRC tissue showed significantly lower C18:0-Cer and C20:0-Cer contents in the tumor•SPT, CerS1, and CerS5 content in CRC tissue was lower than in normal intestinal tissue•A higher circulating content of C16:0-Cer, C18:1-Cer, C20:0-Cer and C24:1-Cer was demonstrated in more advanced colorectal cancer compared to early stages lesions. Here, CERS5 is linked to neoplasm.