游离血红蛋白测定:三波长法和邻联甲苯胺法的比较研究

Determination of free hemoglobin:comparative study of three wavelengths method and orthotolidine method

目的对三波长法和邻联甲苯胺法测定血浆游离血红蛋白(FHb)的准确性、线性、精密度、回收率、稳定性和操作简便性进行比较研究。方法分别用2种方法检测FHb标准品和全血样品中的FHb含量,依次进行准确性、线性、精密度、回收率、稳定性各方面的实验,比较并分析结果。结果三波长法检测得到的FHb标准品含量结果与真实值基本相符,并且直接计算即可得出结果,精密度和回收率都符合方法学要求,检测时呈色稳定;邻联甲苯胺法检测的结果需要外加参照才可获得,吸光度值A与FHb浓度在一定范围内(<600 mg/L)才成线性,精密度不如三波长法,且呈色反应极不稳定。结论三波长法测定FHb结果真实可信,稳定性高,操作简便;邻联甲苯胺法测定FHb含量步骤相对较繁琐,线性范围较窄,注意事项多,稳定性差,比较而言,三波长法的应用前景更为广阔。

Objective To compare the accuracy,linearity,stability,precision,recovery rate,and ease of operation of free hemoglobin（FHb） measurement by using three-wavelength method and orthotolidine method.Methods FHb concentration of FHb standards and whole blood samples were tested with three-wavelength method and orthotolidine method.Then the precision,linearity,recovery,stability were tested and the results analyzed.Results The results obtained by detecting the FHb concentration of FHb standards were close to the real value and the results can be obtained by direct calculation.Recovery and precision were in accordance with methodological requirements.The color development was stable when detection was in progress.The results could be obtained by orthotolidine method only when the reference standard was available.Besides,there was linear relationship between the absorbance（A） and the concentration of FHb only within certain range limits（CFHb600 mg/L）.The precision was no better than the three-wavelength method and the color development was extremely unstable.Conclusion The FHb results obtained by three-wavelength method is easy to operate as well as of high authenticity and stability;the orthotolidine method has obvious disadvantages： tedious operation steps,narrow linear range,many matters needing attention and poor stability.