REMOVAL OF HEPARIN AND PROTAMINE FROM PLASMA

  • 1 January 1976
    • journal article
    • research article
    • Vol. 88  (6) , 922-929
Abstract
A simple chromatographic technique for rapid adsorption of heparin and protamine from plasma samples is described, allowing accurate interpretation of coagulation screening tests and specific clotting factor assays. With the use of columns of ECTEOLA[epichlorohydrintriethanolamine alkali treated]-cellulose, up to 300 U of heparin could be completely adsorbed from a 1 ml plasma sample. When citrated nonheparinized plasma was passed over the ECTEOLA-cellulose columns, the thrombin, prothrombin and partial thromboplastin times were unaffected. Levels of fibrinogen, prothrombin and factors V, VII, VIII, IX and XI average within 90% of control, nonchromatographed samples. When heparinized plasma samples (0.1 and 1.0 U/ml) were passed over columns, heparin was completely removed and the results of the screening tests and the specific factor assays were the same as for the chromatographed nonheparinized samples. Heparinized samples with decreased factor VIII activity also maintained their pretreatment factor VIII activities after heparin removal. Blood samples containing heparin were obtained from 2 patients during open-heart surgery. Following heparin adsorption on ECTEOLA-cellulose columns, factor VIII activity levels remained above 60% during cardiopulmonary by-pass. The presence of protamine sulfate in plasma samples prolonged the prothrombin and partial thromboplastin times while slightly shortening the thrombin time. The protamine effect persisted after ECTEOLA-cellulose but could be removed by a similar column of carboxymethyl-cellulose. The latter resin had no effect on screening tests or on assays of factors VIII or IX activity. The combination of the 2 resins was then used to remove the separate inhibitory effects from heparinized plasma samples to which protamine was added.