Guo Yuanbiao, Guan Min, Du Zheli, Chen Manling, Guo Jianwei, Tu Yi, Zhao Juan, Li Shengli
West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China.
Hua Xi Yi Ke Da Xue Xue Bao. 2002 Jan;33(1):129-31.
To improve the accuracy and precision of the determination of bilirubin, especially direct bilirubin (DB), and the standardization of that as well.
Purified conjugated bilirubin (Bc) and ditaurobilirubin(DTB) and their diazo products were subjected to absorption spectrum analysis. The diazo reaction characters of their calibration solutions were compared by the method of Doumas J-G(TB & DB).
Bc, DTB and their azopigments were found to have the similar absorption spectra with the same lambda max. Their TB standard curves almost superposed together all over. Although the slopes of their DB standard curves were not markedly different ((YBc = 0.00366X + 0.00933, rBc2 = 0.9977, P < 0.01; YDTB = 0.00391X + 0.00023, rDTB2 = 0.9987, P < 0.01; Pb1-b2 > 0.05, n1 = n2 = 5), the DB value measured for Bc differed from that for DTB(n = 5, P < 0.05). In addition, the calibrators made from Bc based different matrices, such as HSA, BSA and human serum, were significantly different in DB/Bc, but no difference was seen among the concentrations. Furthermore, the DB values determined for DTB or Bc increased linearly with the corresponding concentrations, respectively, with no difference between the slopes (YBc = 0.8300XBc + 1.9463, rBc2 = 0.9977, P < 0.01; YDTB = 0.8853XDTB-0.0251, rDTB2 = 0.9986, P < 0.01; n1 = n2 = 5, Pb1-b2 > 0.05).
The results demonstrate that the diazo reaction characters of Bc are identified with those of DTB. However, under the condition of DB, Bc reacts differently from DTB. This study also indicates that as a calibrator of DB based human serum, Bc has the similar constant effect of HCl as serum samples do, so it is a more reliable calibrator to eliminate the matrix effects.
提高胆红素尤其是直接胆红素（DB）测定的准确性和精密度，并实现其标准化。
对纯化的结合胆红素（Bc）和二牛磺胆酸胆红素（DTB）及其重氮产物进行吸收光谱分析。采用Doumas J-G法（总胆红素和直接胆红素）比较它们校准溶液的重氮反应特性。
发现Bc、DTB及其偶氮色素具有相似的吸收光谱，最大吸收波长相同。它们的总胆红素标准曲线几乎完全重叠。虽然它们的直接胆红素标准曲线斜率无明显差异（YBc = 0.00366X + 0.00933，rBc2 = 0.9977，P < 0.01；YDTB = 0.00391X + 0.00023，rDTB2 = 0.9987，P < 0.01；Pb1-b2 > 0.05，n1 = n2 = 5），但Bc测定的直接胆红素值与DTB不同（n = 5，P < 0.05）。此外，基于Bc由不同基质（如人血清白蛋白、牛血清白蛋白和人血清）制成的校准品，其直接胆红素/结合胆红素存在显著差异，但浓度之间无差异。而且，DTB或Bc测定的直接胆红素值分别与相应浓度呈线性增加，斜率之间无差异（YBc = 0.8300XBc + 1.9463，rBc2 = 0.9977，P < 0.01；YDTB = 0.8853XDTB - 0.0251，rDTB2 = 0.9986，P < 0.01；n1 = n2 = 5，Pb1-b2 > 0.05）。
结果表明Bc的重氮反应特性与DTB相同。然而，在直接胆红素条件下，Bc与DTB反应不同。本研究还表明，作为基于人血清的直接胆红素校准品，Bc具有与人血清样品类似的盐酸恒定效应，因此是消除基质效应更可靠的校准品。
