肝素与抗凝血酶Ⅲ对人凝血因子Ⅸ效价检测的影响

Influence of different levels of heparin and antithrombinⅢon human coagulation factorⅨactivity

目的了解不同浓度水平的肝素、抗凝血酶Ⅲ(ATⅢ)或肝素/ATⅢ(1∶1混)对人凝血因子Ⅸ(FⅨ)类制品中FⅨ效价检测的影响。方法向FⅨ或人凝血酶原复合物(PCC)样品中加入不同浓度水平的肝素或肝素/ATⅢ来制备每1 IU FⅨ含0、0.1、0.3、0.5、0.8、1、2、4 IU的肝素或肝素/ATⅢ的样品以及向FⅨ或PCC样品中单独加入ATⅢ时制备每1 IU FⅨ中含0、0.1、0.5、1 IU ATⅢ的样品后,采用一期凝固法测定所制备样品中的FⅨ效价;取适量制备的含不同肝素浓度的样品,分别加入相应量的硫酸鱼精蛋白中和样品中的肝素后再次测定FⅨ效价。观察并分析不同浓度水平肝素、ATⅢ或肝素/ATⅢ对FⅨ效价检测的影响。结果当样品中每1 IU FⅨ的肝素或肝素/ATⅢ含量为0、0.1、0.3、0.5 IU时,FⅨ效价检测结果较一致;当肝素或肝素/ATⅢ含量为0.8、1、2、4 IU时,FⅨ效价检测结果均低于不含肝素样品的检测结果;ATⅢ含量为0、0.1、0.5、1 IU时,样品中FⅨ效价检测一致。结论当FⅨ或PCC制品中每1 IU FⅨ含肝素或肝素/ATⅢ的量≤0.5 IU时,对FⅨ效价的检测结果无影响,可省略加入硫酸鱼精蛋白中和肝素的步骤;>0.5 IU时,宜加入硫酸鱼精蛋白中和肝素后再检测FⅨ效价。

Objective To study the effect of different concentrations of heparin,ATⅢor a mixture of heparin and antithrombinⅢ(ATⅢ)(1∶1)on the activity of human coagulation factorⅨ(FⅨ).Methods The heparin or heparin/ATⅢwith different concentrations were added into human coagulationⅨproducts or human prothrombin complex(PCC)to prepare heparin or heparin/ATⅢsamples,containing 0,0.1,0.3,0.5,0.8,1,2 and 4 IU per unit.ATⅢwith different concentrations were added into FⅨor PCC to prepare ATⅢsamples containing ATⅢ0,0.1,0.5 and 1 IU per unit.The FⅨactivity of the samples prepared was tested by one-stage coagulation method.Then corresponding amount of protamine sulfate were added to neutralize heparin or heparin/ATⅢto detect the FⅨactivity again.Their influence of heparin,ATⅢand heparin/ATⅢwith different concentrations on the activity of FⅨwere analyzed.Results When the content of heparin or heparin/ATⅢwas 0,0.1,0.3 and 0.5 IU per unit of FⅨ,the detection results of FⅨtiter in samples were consistent.When the content of heparin or heparin/ATⅢper unit of FⅨwas 0.8,1,2 and 4 IU,the detection results of FⅨtiter were all lower than those of samples without heparin.When the ATⅢcontent was 0,0.1,0.5 and 1 IU,the FⅨtiter of the samples was consistent.Conclution When the content of heparin or heparin/ATⅢin the product is less than or equal to 0.5 IU per IU of FⅨ,the step of protamine sulfate adding could be omitted as it has little effect on FⅨactivity.When>0.5 IU per IU of FⅨ,however,protamine sulfate adding,to neutralize heparin,is necessary before FⅨactivity testing.