采用冷冻干燥工艺制备地区性凝血因子室间调查品的初步研究

Primary research on freeze drying technology for the preparation of coagulation factor external quality control material in local area

目的采用冷冻干燥工艺制备血浆凝血因子室间调查品。方法对新鲜血浆进行处理加入冻干保护剂,分装后冷冻干燥,使其符合《能力验证样品均匀性和稳定性评价指南》(CNAS-GL03)的要求。将自制的凝血因子调查品分别发至医院进行凝血因子测定。结果采用筛选试验和正交实验,得到的冻干保护剂的最佳配方为海藻糖(1.5%)、甘氨酸(0.5%)、4-羟乙基哌嗪乙磺酸(HEPES,0.2%);分装精度为每瓶的平均分装值为0.569 9 g,范围从0.564 6 g^0.576 7 g,s为0.003 5 g且变异系数(CV)为0.61%。自制冻干品的平均含水量为1.22%,s为0.197%,CV为16%。均匀性采用两种方法评估:(1)F检验:F值为0.06;(2)T检验:P=0.053,所以均匀性符合CNAS-GL03要求;稳定性采用|x珋-y珋|≤0.3σ,即差值的绝对值为0.07<界值1.702符合GL03要求。采用自制冻干品进行室间调查,凝血因子Ⅷ结果为自制冻干品CV%分别为21.3、16.6、25.8(与卫生部临床检验中心的调查结果类似)。结论自制凝血因子的室间调查品可以满足地区性质量控制计划的需要。

Objective To use freeze drying technology for preparing plasma coagulation factor external quality control material,in order to use it into local quality control scheme.Methods Fresh plasma was treated with freeze drying protective agent.Filling the plasma in vials,Germany Christ freeze dryer was used to lyophilize the plasma.The self-made materials were prepared,which should meet the requirements of Guidance on Evaluating the Homogeneity and Stability of Samples Used for Proficiency Testing（CNAS-GL03）.The materials were deliveried into several hospitals for the determination of coagulation factor.Results By screening tests and orthogonal experiments,the optimum freeze drying protective agent＇s formula was trehalose（1.5%）,glycine（0.5%） and HEPES（0.2%）.The average packing bottle precision value was 0.569 9 g（net weight）,ranging from 0.564 6 g to 0.576 7 g,S was 0.003 5 g,and coefficient of variation（CV） was 0.61%.The average moisture of self-made lyophilized product was 1.22%,S was0.197%,and CV was 16%.Homogeneity testing used 2 methods to evaluate（1） F test： F = 0.06;（2） T test： P =0.053 so the homogeneity met the requirements of CNAS-GL03.The stability testing used ｜ x^-- y^-｜ ≤0.3σ,and the absolute value of difference was 0.07,which was〈 critical value 1.702,which met the CNAS-GL03 requirements.By using self-made lyophilized product in external quality control survey,the results of coagulation factor Ⅷ CV% were21.3,16.6 and 25.8（the results were similar to those of the National Center for Clinical Laboratory）.Conclusions The self-made coagulation factor external quality control material can meet the requirements of external quality control scheme in local area.