The invention discloses a cross-linked
hemoglobin preparation method, wherein the preparation method comprises the following steps: a, diluting deoxidized
hemoglobin, and adding the diluted deoxidizedhemoglobin into an
anaerobic reactor; b, arranging an atomizer at the top of the
anaerobic reactor, adding a cross-linking agent into the reactor through the atomizer, and carrying out a cross-linking reaction; and c, adding a
reducing agent to terminate the cross-linking reaction, and removing the unreacted cross-linking agent and the terminating
reducing agent in an
ultrafiltration liquid changing manner to obtain the cross-linked
hemoglobin. The cross-linked hemoglobin prepared by the preparation method contains low contents of macromolecules (greater than 500 kD) and micromolecules (lessthan 64 kD); by adding the cross-linked hemoglobin into organ perfusate, the phenomena of organ
acidosis and lysosomal
enzyme activation adverse consequences caused by lack of an
oxygen carrying agentin conventional perfusate and
cell damage caused by excessive free radicals generated during reperfusion can be solved, and the perfusate is quite simple and convenient to prepare, store, transport and use.