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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 compare the plasma free hemoglobin with three wavelength colorimetric method and compare with o-toluidine method to explore the feasibility of this method. Methods: Different concentrations of hemoglobin (Hb) standard application solution were added to the prepared Tris-HCl buffer diluent tube, after mixing, respectively, at 380 nm, 415 nm, 450 nm colorimetry, calculated according to the formula Hb Content, and with o-toluidine method for comparison. Results: The linear range of this method was (0 ~ 800) mg / L, the minimum detectable limit was 2.25 mg / L. The CV of two different concentrations of Hb solution were 3.28% and 1.52%, respectively. The results of inter-assay repeated test (CV) were 5.44% and 3.51%, respectively. The recovery results were 98.1% , 102.6%; control experiment regression equation Y = 0.989 (1.023X + 0.218), γ = 0.989. Conclusion: The results of three-wavelength colorimetric assay showed good repeatability, high accuracy, good linearity, simple operation and no contamination of the reagent used, which provided a better method selection for detecting plasma free hemoglobin.