Experimental study of Genipin cross-linked hemoglobin isolated from fetal

Abstract

Objective To achieve effective polymerization rate (molecular weight of 64-600 KD) ≥70% and average molecular weight of 100-300 KD across the way of Gennipin (Genipin, Gnp) crosslinked hemoglobin isolated from fetal (hemoglobin isolated from fetal, HbF). Methods HbF was used as raw material, and Gnp as crosslinker. Orthogonal test involving four factors as hemoglobin concentration (HbC), molar ratio of Gnp to HbF, reaction time and reaction pH, and three levels were selected (n=3) to determine better matching condition. Three batches of Gnp-HbF samples were prepared under the optimal conditions, and the effective polymerization rate, dimer, average molecular weight, the content of methemoglobin (MetHb) and P50 were determined during the preparation process. Results The optimal matching conditions for Gnp-HbF obtained by orthogonal test were HbC: 80g/L, molar ratio of Gnp to HbF: 7∶1, reaction time: 18 h, pH: 8.5, with effective polymerization rate at (73.03±2.03)%, dimer (26.97±2.03)%, average molecular weight (167.70±28.40)KD, the content of methemoglobin(2.60±0.17)% and P50 (13.13±1.02) mmHg. Conclusion Gnp can effectively cross-link HbF, and the obtained key indicators of Gnp-HbF products meet the requirements, which reduces the preparation difficulty for the subsequent process.

Keywords

• genipin(gnp)
• hemoglobin
• orthogonal test/four factors
• hemoglobin-based oxygen carrier