Lu K C, Gooding K M, Regnier F E
Clin Chem. 1979 Sep;25(9):1608-12.
Evaulation of the severity of jaundice in the neonate may be determined by measuring the reserve binding capacity of serum proteins for free bilirubin. Determination of protein-bound bilirubin has been labor intensive, necessitating multiple runs on gel-permeation chromatography columns or, more recently, enzyme assays or fluorescence quenching analysis. We present a method for quantitation of free bilirubin and of bilirubin-binding capacity of serum by liquid chromatography. A gel-permeation column binds free bilirubin while allowing passage and quantitation of protein-bound bilirubin. Subsequent injection of a desorbing agent releases the adsorbed bilirubin from the column, permitting quantitation of free bilirubin. Bound and free serum bilirubin may be determined directly in less than 15 min using 10 microL of serum. The binding of bilirubin to neonatal serum is seen to be quite different from the binding to adult serum. Ion-exchange chromatography of adult and neonatal serum samples shows that their protein profiles are radically different. This difference probably accounts for the binding characteristics.
评估新生儿黄疸的严重程度可通过测量血清蛋白对游离胆红素的储备结合能力来确定。测定结合胆红素一直是劳动强度大的工作，需要在凝胶渗透色谱柱上进行多次分析，或者最近采用酶法或荧光猝灭分析。我们提出一种通过液相色谱法定量游离胆红素和血清胆红素结合能力的方法。凝胶渗透柱结合游离胆红素，同时允许结合胆红素通过并进行定量。随后注入解吸剂可从柱上释放吸附的胆红素，从而实现游离胆红素的定量。使用10微升血清可在不到15分钟内直接测定结合和游离血清胆红素。胆红素与新生儿血清的结合与与成人血清的结合明显不同。成人和新生儿血清样本的离子交换色谱显示，它们的蛋白质谱有根本差异。这种差异可能解释了结合特性。
