KHC_3法测定胆红素试剂的研制

目的:探讨建立一种新型KHC3法测定血清总胆红素(TBIL)和直接胆红素(DBIL)的新方法。方法:采用化学氧化法测定血清胆红素反应到达终点时的吸光度,与标准品相比较,求得标本中总胆红素和直接胆红素的浓度。结果:精密度试验TBIL批内高低浓度CV值为0.78%、1.44%;DBIL为0.96%、1.85%。TBIL批间为3.36%、2.66%及3.79%;DBIL批间为4.03%、3.25%及2.96%。线性范围:TBIL可达515μmol/L;DBIL可达171μmol/L;总胆回收率98.4%～105.3%,平均102.1%;DBIL回收率为98.5%～101.5%,平均为99.7%。与其他相关试剂比较TBILγ=0.9954～0.9994;DBILγ=0.9950～0.9985。溶血、脂浊和维生素C干扰试验表明:标本中Hb小于10g/L,TG小于7.55mmol/L,VitC小于0.1g/L,对TBIL和DBIL的测定基本无干扰。结论:本试剂方法简便,结果准确、线性范围宽,试剂稳定,价格低廉,完全可用于检测临床标本。

Development of the reagents with KHC3 method for measuring serum bilirubin

Objective To establish a new KHC3 method for measuring serum bilirubin.Methods KHC3 method was applied for detecting serum total bilirubin(TBIL) and cholebilirubin(DBIL) concentrations according to the absorbance changes,and the results were analysed comparing with the standard controls.Results The intraassay CVs of TBIL were 0.78% and 1.44%(n=15),with the mean values of 22.3 and 258.6μmol/L respectively,while that of DBIL were 0.96% and 1.85%(n=15),with the mean values of 7.5 and 116.6μmol/L respectively.The inter-assay CVs of TBIL were 3.36%,2.66% and 3.79%(n=20),with the mean values of 14.6,164.5 and 483.4μmol/L respectively,while that of DBIL were 4.03%,3.25% and 2.96%(n=20),with the mean values of 4.4,87.2 and 153.6μmol/L respectively.The linear ranges were up to 515μmol/L for TBIL and 171μmol/L for DBIL respestively.The average recoveries were 102.1% for TBIL and 99.7% for DBIL.The nitrite method was well correlated with Vanadate method and Diazo method.When hemoglobin level in serum was under 10g/L,Triglycerides under 7.55 mmol/L,or Vitamin C under 0.1g/L,the results showed no significant interference for measuring bilirubin.Conclusion This new KHC3 method is simple and stable,and it can be used in routine detections of serum total TBIL and DBIL.