Studies on the purification of an anti-trypanosomal factor (ATF- II) from the bacterial species, Pseudomonas fluorescens, were continued. It was necessary to modify the purification procedure used previously in order to obtain pure fractions that were suitable for NMR analysis. As before, we used high-speed countercurrent chromatography (CCC) employing the Ito multilayer coiled column but in the high pressure chromatography system (HPLC) we employed both preparative and analytical carbohydrate analysis columns to obtain the NMR fractions. Work is in progress to obtain sufficient amounts of pure compound for these studies. The determination of chemical structure is essential before studies on synthesis are initiated and pharmacological and toxicological properties are examined employing experimental infections with Trypanosoma cruzi.