冷沉淀凝血因子融化后储存温度和时间对凝血因子的影响

Effect of storage temperature and time on coagulation factor after cryoprecipitated antihemophilic factor melting

目的 通过研究比较冷沉淀凝血因子融化后不同的储存温度和时间对凝血因子的影响,为行业标准的制定提供参考依据。方法 2021年6月—2023年5月,每月抽检4袋共96袋冷沉淀凝血因子,当月抽检并及时检测。冷沉淀凝血因子37℃水浴融化后对轻中度脂血予以标记,每袋冷沉淀凝血因子和2个50 mL转移袋用无菌接驳机分装成2袋2组各20 mL,1组放置4℃冰箱,另1组放置22℃水浴箱中,放置时间均为0 h、4 h、8 h、12 h、24 h、48 h,然后再各自时间无菌取样2 mL放入试管中,用加样枪在另1试管中加入样品1 mL和缓冲液3 mL混匀上机进行检测。随机选取60袋无轻中度脂血冷沉淀凝血因子的实验数据,采用SPSS21.0进行统计分析。结果 融化后的冷沉淀凝血因子贮放0 h、4 h、8 h、12 h、24 h、48 h检测2组凝血因子平均含量及增长率:(1)储存4℃,Ⅷ因子含量依次为118.62、111.57(-5.95%)、105.51(-11.05%)、103.30(-12.92%)、94.35(-20.46%)、83.25(-29.82%)IU/袋;储存22℃,Ⅷ因子含量依次为118.62、112.69(-5.00%)、111.41(-6.08%)、109.01(-8.10%)、101.55(-14.39%)、92.75(-21.81%)IU/袋,2组储存结果比较,在4℃24 h和22℃48 h,Ⅷ因子含量均有显著统计学意义(P<0.01),且在22℃储存,Ⅷ因子衰减速度更慢;(2)储存4℃,Ⅴ因子含量依次为41.19、41.31(0.29%)、40.52(-1.64%)、40.27(-2.23%)、39.05(-5.19%)、36.99(-10.21%)IU/袋;储存22℃,Ⅴ因子含量依次为41.19、41.71(1.25%)、42.54(3.28%)、41.94(1.80%)、39.21(-4.80%)、35.64(-13.48%)IU/袋,2组储存结果比较,在4℃48 h和22℃48 hⅤ因子含量分别有统计学意义(P<0.05)和有显著统计学意义(P<0.01),且在22℃储存,Ⅴ因子衰减速度更快;(3)储存4℃,Fbg含量依次为268.86、268.17(-0.26%)、262.46(-2.38%)、270.50(0.61%)、267.52(-0.50%)、261.92(-2.58%)mg/袋;储存22℃,Fbg含量依次为268.86、265.86(-1.12%)、264.12(-1.77%)、265.89(-1.11%)、266.04(-1.05%)、261.04(-2.91%)mg/袋,2组各时间段与0 h含量比对均无统计学意义(P>0.05)。结论 冷沉淀凝血因子融化后,凝血因子随储存时间延长而下降,尤其Ⅷ因子含量下降最明显,Ⅴ因子次之,Fbg基本不变。2组储存比较,22℃储存Ⅷ因子衰减速度更慢,Ⅴ因子衰减速度更快。冷沉淀凝血因子融化后应尽快输注;如果延迟不可避免,若延迟时间<12 h,采用4℃保存;若延迟时间介于12~24 h,采用22℃保存为宜。

Objective To study and compare the effects of different storage temperature and time on coagulation factor after cryoprecipitated antihemophilic factor(CAF)melting,and to provide reference for the establishment of industry standards.Methods From June 2021 to May 2023,a total of 96 bags of CAF were sampled in 4 bags per month,and timely detected in the same month.After the CAF was melted in a 37℃water bath,the mild to moderate lipemic blood was labeled.Each bag of CAF and two 50 mL transfer bags were divided into two bags and two groups of 20 mL each using a sterile adapter.One group was placed in a 4℃refrigerator and the other in a 22℃water bath for 0 h,4 h,8 h,12 h,24 h and 48 h.Then 2 mL of aseptic sample was taken separately and put into the test tube,and 1mL of sample and 3 mL of buffer were added into the other test tube with the sampling gun and mixed on the machine for testing.The experimental data of 60 bags without mild to moderate lipemic blood cryoprecipitation and coagulation factor were randomly selected and statistically analyzed by SPSS21.0.Results After melting,CAF was stored for 0 h,4 h,8 h,12 h,24 h and 48 h to detect the average content and growth rate of coagulation factor in the two groups:1)Storage at 4℃,factorⅧcontent was 118.62,111.57(-5.95%),105.51(-11.05%),103.30(-12.92%),94.35(-20.46%)and 83.25(-29.82%)IU/bag,respectively;Storage at 22℃,the factorⅧcontent was 118.62,112.69(-5.00%),111.41(-6.08%),109.01(-8.10%),101.55(-14.39%)and 92.75(-21.81%)IU/bag,and the storage results of the two groups were compared.At 24 h at 4℃and 48 h at 22℃,the content of factorⅧhad significant statistical significance(P<0.01),and when stored at 22℃,the decay rate of factorⅧwas slower;2)When stored at 4℃,the content of factor V was 41.19,41.31(0.29%),40.52(-1.64%),40.27(-2.23%),39.05(-5.19%)and 36.99(-10.21%)IU/bag,respectively;Stored at 22℃,the factor V content was 41.19,41.71(1.25%),42.54(3.28%),41.94(1.80%),39.21(-4.80%)and 35.64(-13.48%)IU/bag,respectively.Comparison of storage results between the two groups showed that the content of factor V was statistically significant(P<0.05)and significantly significant(P<0.01)at 4℃48 h and 22℃48 h,respectively,and the decay rate of factor V was faster when stored at 22℃;3)When stored at 4℃,the Fbg content was 268.86,268.17(-0.26%),262.46(-2.38%),270.50(0.61%),267.52(-0.50%)and 261.92(-2.58%)mg/bag,respectively;Stored at 22℃,the Fbg content was 268.86,265.86(-1.12%),264.12(-1.77%),265.89(-1.11%),266.04(-1.05%)and 261.04(-2.91%)mg/bag,respectively.There was no statistical significance between the 2 groups and the original 0 h content in each time period(P>0.05).Conclusion After CAF melting,coagulation factor decreased with the extension of storage time,especially the decrease of factorⅧ,followed by factor V,while Fbg basically unchanged.Comparison between the two groups showed that,factorⅧdecay rate is slower,factor V decay rate is faster of storage at 22℃.CAF should be transfused as soon as possible after melting.If the delay is unavoidable,for the delay time less than 12 h,storage at 4℃is recommended,fot the delay time more than 12 h and less than 24 h,storage at 22℃is recommended.