EP 1 211 261 A2

EP 1 211 261 A25[0220] In addition, evaluation of the blood-substitute, produced according to this method, showed that the bloodsubstitutealso satisfactorily met a general safety test. In the general safety test, 2 mice and 2 guinea pigs were injectedwith blood-substitute, after which the test animals were monitored for 7 days. None of the animals injected with bloodsubstituteexperienced a weight gain, a weight loss or death, thereby demonstrating the general safety of the bloodsubstitute.Example 41015202530354045Stability Analysis of Stable Polymerized Hemoglobin Blood-Substitute[0221] The stability of polymerized hemoglobin blood-substitute, produced according to the method of this invention,was evaluated over a period of 24 months at various storage temperatures. The specific storage temperatures evaluatedwere 2-8°C, room temperature (about 25°C) and 37°C. For storage at 2-8°C and at room temperature (RT), thestability results (provided in Table VI) show that the blood-substitute was stable for two years with only de minimischanges in the composition of the blood-substitute. Further, for storage at 37°C, the blood-substitute was stable forover one year. Blood-substitute stored at 37°C for 18 months, however, showed < 5 particles/ml at 25 µ and more than11% Hb with a molecular weight greater than 500,000 Daltons.[0222] In these analyses, total hemoglobin, methemoglobin and oxyhemoglobin concentrations were determined ona Co-Oximeter Model #482, from Instrumentation Laboratory, Lexington, MA.Table VITest and Specification Initial 24-Month Storage at2-8°C24-Month Storage atRT12-Month Storage at37°CPackage Integrity Meets Test Meets Test Meets Test Meets TestTotal Hb 13±1 g/dl 12.6 12.8 12.8 12.8MetHb 10% 3.1 1.8 2.0 1.4Glutaraldehyde