Glycoconjugates from five trypanosomatid genera-Crithidia, Herpetomonas, Endotrypanum, Leishmania, and Trypanosoma-were extracted with Triton X-114 and analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis followed by periodic acid-Schiff staining. Most of the glycoconjugates were detected in the hydrophobic phase, indicating the presence of anchored glycoconjugates. All the trypanosomatids expressed a glycoconjugate with a low molecular weight (below 20 kDa) in this phase. In each species, however, a characteristic and specific pattern of glycoconjugates was also observed in both phases. In the hydrophobic phase: 14-29 kDa glycoconjugates in C. guilhermei; 24-70 kDa in C. fasciculata, C. luciliae, E. schaudinni, and T. cruzi Y and G strains; 45-66 kDa in C. oncopelti and H. samuelpessoai; above 36 kDa in T. dionisii; 20-24 kDa, 36-45 kDa, and 70 kDa in L. tarentolae and T. mega. In the hydrophilic phase, typical glycoproteins were observed in some trypanosomatids: 60 kDa in T. mega and T. cruzi Y strain; 70 kDa in H. samuelpessoai; 66 kDa in C. oncopelti; 20-70 kDa in C. luciliae, These findings suggest that Triton X-114-extracted glycoconjugates could be useful markers for trypanosomatid taxonomy,