An Application of Orthogonal Test in Optimizing Lyophilized Protective Agent of Human Plasma Fibrinogen
Objective Using fibrinogen in human plasma as the research object,this paper explored how to select protective agents during vacuum freeze-drying,and evaluated freeze-dried products,so as to establish an efficient protective agent screening method by orthogonal test.Methods Mannitol,glycine and sodium azide were selected by the primary screening test and were added to plasma lyophilized by orthogonal test with high and low concentrations,respectively.The primary and secondary relationship and final concentration of each protective agent were determined by range analysis and variance analysis.In addition,sterility,quality,homogeneity and stability of freeze-dried products were evaluated to verify the applicability of the selected protective agents.Results The results showed that there was an interaction between mannitol and sodium azide by using orthogonal test and drawing binary graph.Anova was superior to range method and the interaction between mannitol and sodium azide and sodium azide were the main factors,while the rest were the secondary factors.The best combination of protective agents was:mannitol 5％,glycine 0.5％,and sodium azide 0.1％.The sterility,appearance,water content and dissolution of freeze-dried products were all in line with the requirements.The freeze-dried product was homogeneous,and the CV bottle interval was 2.7％,which were stable within 8 hours after dissolution,stable within 14 days after transportation at 2-8℃,and stable for 6 months after freezing at-20℃.Conclusion Orthogonal test method can be used to efficiently and accurately identify the best combination in the selection of multi-factor lyophilized protective agents,and has a wide range of application values.
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