Glass wool, hydrophilic cotton wool, non-electrically charged BIO-GEL P2 and common tissue paper columns were used to purify trypomastigotes from a mixed Trypanosoma cruzi population grown in axenic culture medium. With all these columns, highly purified (up to 98%) trypomastigote preparations were obtained. Trypomastigote yields from cotton wool, BIO-GEL P2 and common tissue paper columns were not as high as from glass wool columns, from which yields varied from 69 to 80%. Purification on glass wool did not affect trypomastigote infectivity or virulence. Dead trypomastigotes could not be purified on glass wool columns. A glass-adherent amphiphilic peptide of 45 kDa, present in the cell membrane, was isolated from epimastigote but not from trypomastigote preparations.