Studies on the quality control of stroma-free hemoglobin

Analytical methods for phospholipids and blood group antigen were developed using HPLC-UV and EIA, respectively, for the quality control of SFH. These methods had sensitivities superior to those of conventional assays. In addition, the HPLC method could isolate PS, which might play an essential role in blood coagulation. The above analytical methods were applied to evaluate three representatives SFH preparation methods, 36,000xg centrifugation followed by the filtration with a 0.22 um filter, ultrafiltration with a 100 kd membrane, and filtration with the BMM-40 nm membrane. Both the BMM method and the ultrafiltration method were shown to remove more than 99.7% of phospholipids, and more than 99.99% of blood group antigen. On the other hand, the centrifugation preparation still contained much of both components. The BMM filter was originally developed as a virus removal filter. The virus removal efficiency of the BMM-40 nm was examined with a genetic technique, the PCR method. The BMM-40 nm was demonstrated to remove HB viruses with an efficiency of eight log or more. These results indicate that the BMM is a useful filter to prepare a stroma-free, virus-reduced hemoglobin solution.