A hemoglobina humana (Hb) vem sendo usada como pon to de partida para o desenvolvimento de substitutos para o sangue. Antes de ser submetida a reações de modifica ção química, a Hb precisa ser obtida com alto grau de pureza. Neste trabalho, concentrados de hemácias foram tratados por métodos convencionais para a obtenção de amostras de Hb, sob a forma de carbonil-hemoglobina (HbCO). A técnica de cromatografia de troca iônica foi usada com o objetivo de verificar-se a eficiência de resinas aniônicas na eliminação de fosfolipídeos da parede celular, ainda residuais nas soluções de HbCO. Experimentos fo ram realizados com duas resinas de troca aniônica, uma fraca e outra forte. Os extratos orgânicos da solução de HbCO purificada convencionalmente e da solução purificada adicionalmente por meio de cromatografia em resina de troca aniônica forte, AG MP-1, foram analisados por cromatografia líquida de alta pressão (HPLC). Os re sultados mostraram que a resina AG MP-1, sob as condi ções utilizadas, foi capaz de reter grande parte do total de fosfolipídeos pesquisados.

Human hemoglobin (Hb) has been used to develop blood substi tutes. Before being submitted to chemical modification reactions, purification procedures should be designed in order to obtain stroma-free hemoglobin. In the present work, red blood cells were treated with CO gas to convert Hb to carbonyl hemoglobin, and then conventional purification methods were applied. Ion-ex change chromatography was used to verify the efficiency of an ionic resins to eliminate stroma contaminants. Weak and strong anion-exchangers were used. Better results were obtained with the strong anion-exchanger AG MP-1. Some modifications of the conventional purification procedure were also adopted, such as microfiltration of the hemolysate. Hemoglobin concentrations were determined by spectrophotometry. High-performance liq uid chromatography was used to quantify the contents of phos phatidylethanolamine, phosphatidylcholine and sphingomyelin in the organic extracts from HbCO solutions prepared in the usual way and from HbCO solutions submitted to further puri fication by anion-exchange chromatography with the AG MP-1 resin. The results showed that the AG MP-1 resin was able to adsorb 94% of the total content of phospholipids present in the HbCO solution purified by the conventional method followed by microfiltration.