游离血红蛋白浓度的定量检测方法及其在失血性休克复苏中的应用

Quantitative determination of free hemoglobin and its application in resuscitation for hemorrhagic shock

目的研究全血和血液制品中游离血红蛋白(free hemoglobin,FHb)浓度的定量检测方法及其在大鼠失血性休克模型输血复苏时红细胞溶血的评价。方法通过检测血浆本底比色反应后的吸收光谱和制备系列浓度FHb标准液与光密度值的标准曲线,进行FHb检测的方法学验证;建立Wistar大鼠的失血性休克模型,随机分为5组,失血40%后,A组仅输血液,B、C、D、E组根据动物体质量分别输注含不同浓度FHb(10,20,30,40 mg/kg)的血液;在基线水平和复苏后1 h抽取动物股动脉血液,测定对照组和实验组动物血液的FHb浓度值,并进行血气分析。结果 FHb检测装置及方法的浓度范围为0～2 g/L,日内和日间精密度分别小于3.6%和6%。将该方法应用于复苏前后血液中FHb浓度的定量检测,结果显示,各组动物FHb的检测结果在复苏前后均有统计学意义(P<0.01),含不同浓度FHb的血液与液体进入体内后1 h,pH水平得到了纠正,并发现复苏前后FHb测定结果与pH的变化呈线性相关(R2=0.905,P<0.01)。结论建立一种FHb定量检测的方法,可用于院前输血治疗时全血或血液制品质量安全的快速评估。

Objective To develop a quantitative determination method for free hemoglobin（FHb） concentration in whole blood and blood products,and to evaluate the blood transfusion in rats with hemorrhagic shock.Methods A hemorrhagic shock model was reproduced with 40% blood loss.Twenty-five Wistar rats were randomly divided into five groups： Group A treated by blood（n=5）,Group B treated by blood with 10 mg/kg FHb（n=5）,Group C treated by blood with 20 mg/kg FHb（n=5）,Group D treated by blood with 30 mg/kg FHb（n=5）,and Group E treated by blood with 40 mg/kg FHb（n=5）.In baseline and post-infusion 1 h,FHb concentrations of each group were measured,and blood gases of animals were analyzed.Results The detection range was from 0 to 2 g/L.The coefficient of variation（CV） of inter-day variability and intra-day variability were less than 3.6% and 6.0%,respectively.The method was applied to quantitative detection of FHb concentration before and after blood transfusion.The results showed that FHb values of animals in each group were statistically significant（P0.01） before and after blood transfusion.When animals were infused by blood with different FHb concentrations,pH levels were corrected for 1 h,and the changes of pH were in a linear correlation with FHb concentrations（R2=0.905,P0.01）.Conclusion A simple and accurate measurement method for FHb concentration is established,which can be applied to rapidly assess the quality and safety of whole blood or blood products on blood transfusion in the pre-hospital stage.