FLUOROMETRIC STUDY OF THE PARTITION OF BILIRUBIN AMONG BLOOD COMPONENTS - BASIS FOR RAPID MICROASSAYS OF BILIRUBIN AND BILIRUBIN BINDING-CAPACITY IN WHOLE-BLOOD
FLUOROMETRIC STUDY OF THE PARTITION OF BILIRUBIN AMONG BLOOD COMPONENTS - BASIS FOR RAPID MICROASSAYS OF BILIRUBIN AND BILIRUBIN BINDING-CAPACITY IN WHOLE-BLOOD
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DOI：
10.1016/0003-2697(79)90105-2
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发表时间：
1979-01-01
影响因子：
2.9
通讯作者：
FLORES, J
中科院分区：
文献类型：
--
作者：
LAMOLA, AA;EISINGER, J;FLORES, J
胆红素与血液中的许多位点结合，最强的结合是与白蛋白上的单个位点。白蛋白上的次要位点、其他血浆蛋白上的大多数位点和红细胞膜上的位点对胆红素的亲和力最多只有 1/100。胆红素与红细胞中的 Hb 结合的有效亲和力小于初级白蛋白位点的 1/1000。基本上，血液中存在的唯一发出荧光的胆红素是与初级白蛋白位点结合的胆红素。血液中几乎所有其他胆红素都会发出荧光，但产量不超过其 1/50。使用正面技术可以对全血进行定量荧光测定。可以通过这种方式确定与初级白蛋白位点结合的胆红素的浓度。可以通过用胆红素滴定样本来类似地测定血液样本的白蛋白结合能力。非离子去污剂十二烷基二甲胺氧化物（DDAO）可清除血液中所有部位的胆红素，并且由于DDAO胶束中的胆红素呈荧光，因此在样本中添加DDAO后，可以通过荧光法测定血液总胆红素。这种去污剂方法还可以轻松测定红细胞结合胆红素。这些针对总血胆红素、白蛋白结合胆红素和白蛋白结合能力的荧光测定法简单、快速，并且使用的血液量非常小。它们对于新生儿黄疸的研究及其临床治疗具有重要价值。
Bilirubin bounds to many sites in blood, the strongest binding being to a single site on albumin. Secondary sites on albumin, most sites on other plasma proteins and sites on erythrocyte membranes had affinities for bilirubin that were at most 1/100 as great. Bilirubin binds to Hb in red cells with an effective affinity that is less than 1/1000 that of the primary albumin site. Essentially the only bilirubin present in blood which fluoresced was that bound to the primary albumin site. Almost all the other bilirubin in blood fluoresced with a yield no more than 1/50 as large. Quantitative fluorometry of whole blood was possible using the front-face technique. The concentration of bilirubin bound to the primary albumin site could be determined in this way. The albumin binding capacity of a blood specimen could be similarly assayed upon titration of the specimen with bilirubin. The nonionic detergent dodecyldimethylamine oxide (DDAO) scavenged bilirubin from all sites in blood, and, since bilirubin was fluorescent in DDAO micelles, the total blood bilirubin could be assayed fluorometrically after addition of DDAO to the specimen. This detergent method also allowed facile assay of red-cell-bound bilirubin. These fluorometric assays for total blood bilirubin, albumin-bound bilirubin and albumin binding capacity were simple and rapid and used very small volumes of blood. They should be of great value in the research on neonatal jaundice and in its clinical management.