三波长比色法测定血浆游离血红蛋白的探讨

Study on determination of plasma free hemoglobin by three-wavelength colorimetry

目的:应用三波长比色法测定血浆游离血红蛋白,并与邻-甲联苯胺法相比较,探讨该方法的可行性。方法:将不同浓度的血红蛋白(Hb)标准应用液分别加入到配制好的Tris-HCl缓冲稀释液的试管中,混匀后分别在380 nm、415 nm、450 nm比色,按公式算Hb的含量,并与邻-甲联苯胺法进行对照。结果:该法线性范围为(0～800)mg/L,最低检出限为2.25 mg/L。2种不同浓度的Hb应用液批内重复性试验(CV)结果分别为3.28%、1.52%,批间重复性试验(CV)结果分别为5.44%、3.51%;回收率实验结果分别为98.1%、102.6%;对照实验回归方程Y=0.989(1.023X+0.218),γ=0.989。结论:三波长比色法实验结果重复性好、准确度高、线性好,且操作简单,所用试剂无污染,为检测血浆游离血红蛋白提供较好的方法选择。

Objective：To determine the concentration of plasma free hemoglobin by three-wavelength colorimery,and discuss the feasibility of this method by comparison with method of o tolidine.Methods：Different concentrations of hemoglobin（Hb） were respectively added to the test tube with standard Tris-HCl buffer solution.The absorbance of those mixture were determined at wavelength of 380 nm,415 nm and 450 nm respectively.The concentrations of Hb calculated by the formula and compared with method of o tolidine.Results：The linear range of this method was（0～800） mg/L,the determination limit was 2.25 mg/L;Two kinds of Hb application liquids were tested repeatedly,the intraassay coefficients of variation（CV） were 3.28% and 1.52% respectively,the interassay coefficients of variation（CV） were 5.44% and 3.51% respectively;the results of recovery were 98.1% and 102.6% respectively;the equation of control test was Y= 0.989（1.023 X ＋ 0.218）,γ=0.989.Conclusion：The operation of three-wavelength colorimery was simple and the reagents were non-polluting.This method can be used to determine plasma free hemoglobin with good repeatability,high accuracy and good linearity.