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中文摘要：
目的采用正交试验法优化凝血酶原复合物（Prothrombin complex concentrate,PCC）的制备工艺。方法取DEAE-Sephadex A-50凝胶平衡体系中的柠檬酸钠、氯化钠、pH值3因素,每因素取3水平设计正交试验;称取9等份DEAE-Sephadex A-50凝胶干粉,分别采用试验设计的平衡体系进行平衡;平衡后的凝胶分别与等量健康人去冷沉淀混合血浆混合,制备凝血酶原复合物浓缩物;测定浓缩物蛋白浓度及凝血因子Ⅱ、Ⅶ、Ⅸ、Ⅹ活性,获得4种凝血因子比活;通过正交试验直观分析及直接对比法确定较优及较差比活制备条件,对确定的条件进行验证;测定平衡体系电导率,分析正交试验中平衡体系电导率与4种凝血因子比活的相关性。结果正交试验直观分析法与直接对比法确定的凝血因子Ⅱ、Ⅶ、Ⅸ、Ⅹ较优比活凝胶平衡条件一致：柠檬酸钠0.005~0.012 mol/L,氯化钠0.06~0.10 mol/L,pH 6.9~7.2;两种方法确定的较差平衡条件不同,分别为：柠檬酸钠0.012~0.02 mol/L、氯化钠0.1~0.14 mol/L,pH 7.2~7.5及柠檬酸钠0.005~0.012 mol/L,氯化钠0.1~0.14 mol/L,pH 7.2~7.5,经实验验证,两种方法确定的较优条件比活均优于较差条件,但两种较差条件中,直接对比法确定的条件更符合实际。平衡液电导率与4种凝血因子比活之间不存在线性关系,但在不同电导率条件下,4种凝血因子比活有较大波动,电导率9.67~10.34 ms/cm,4种凝血因子比活均较高;电导率12.16~12.90 ms/cm,4种凝血因子比活均较低。结论正交试验可用于凝血酶原复合物制备工艺优化,但不同分析方法结果有一定差异。平衡体系电导率对凝血酶原复合物比活有一定影响,在工艺优化时应予注意。
英文摘要：
Objective To optimize the preparation procedure for prothrombin complex concentrate（PCC） by orthogonal test.Methods Three factors in DEAE-Sephadex A-50 gel equilibration system,i.e.sodium citrate,sodium chloride and pH value,were selected for orthogonal test at three levels.Nine aliquots of DEAE-Sephadex A-50 gel powder were equilibrated by the designed equilibration system.The gels were mixed with equal volumes of pooled cryoprecipitate supernatant of healthy human plasma to prepare PCC which was determined for protein concentration and activities of coagulation factors Ⅱ,Ⅶ,Ⅸ,Ⅹ,based on which the specific activities of the four coagulation factors were calculated.The better and worse equilibration conditions for specific activities of the four coagulation factors were determined by heuristic analysis and direct comparison in orthogonal test,and were further verified.The conductivity of equilibration system was determined,of which the relationship to specific activities of four coagulation factors was analyzed.Results The better equilibration conditions for specific activities of four coagulation factors determined by heuristic analysis and direct comparison were in agreement： sodium citrate 0.005 ~ 0.012 mol / L,sodium chloride 0.06 ~ 0.10 mol / L,pH 6.9 ~ 7.2.However,the worse equilibration conditions were different,which were sodium citrate 0.012 ~ 0.02 mol / L,sodium chloride 0.10 ~ 0.14 mol / L,pH 7.2 ~ 7.5 by heuristic analysis,and sodium citrate 0.005 ~ 0.012 mol / L,sodium chloride 0.10 ~ 0.14 mol / L,and pH 7.2 ~ 7.5 by direct comparison.Verification test showed that the specific activities of four coagulation factors under the better equilibration conditions determined by the two methods were higher than those under the worse conditions.However,the worse conditions determined by direct comparison were more consistent with those in actual fact.The conductivity of equilibration buffer showed no linear relationship to the specific activities of four coagulation factors.However,the specific ac
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