Abstract
Two analytical methods; high-performance liquid chromatography (HPLC) for phospholipids detection and enzyme immunoassay (EIA) for blood group antigen detection, were developed for the purpose of quality control of stroma-free hemoglobin (SFH). By HPLC four major phospholipids were separated clearly. The minimum detection levels of phosphatidylserin, phosphatidylethanolamine, phosphatidylcholine and sphingomyelin were 0.1, 0.2, 0.03 and 0.2μg, respectively. EIA had good reproducibility and the minimum detection level of blood group antigen was 6ng/ml. the ultrafiltration method with a 100kd membrane removed stroma effectively, as shown the prepared SFH contained only 1.4μg/ml of phospholipids and 12ng/ml of antigen. The removal efficiency of the stroma by the BMM methods was similar to the ultrafiltration method, whereas SFH prepared by 36, 0000xg centrifugation method contained 4.0μg/ml of phospholipids and 0.2μg/ml of antigen. As most conventional assays can not measure these low level of stroma, it is indicated that HPLC and EIA are useful to evaluate stroma content in SFH.
