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Liver disorders and gallstones
Published in Martin Andrew Crook, Clinical Biochemistry & Metabolic Medicine, 2013
Bilirubin monoglucuronide passes to the canalicular surfaces of the hepatocytes, where, after the addition of a second glucuronate molecule, it is secreted by active processes into the bile canaliculi. This process is largely dependent on the active secretion of bile acids from hepatocytes. These energy-dependent steps are the ones most likely to be impaired by liver damage (hypoxia and septicaemia) and by increased pressure in the biliary tract. Other anions, including drugs, may compete for binding to ligandin, thus impairing bilirubin conjugation and excretion. Novobiocin inhibits glucuronyl transferase, thus exacerbating unconjugated hyperbilirubinaemia. Bilirubin is often assayed by the Van den Bergh reaction, which allows conjugated (direct-reacting) and unconju-gated (indirect-reacting) bilirubin to be distinguished.
Structure, Photochemistry, and Organic Chemistry of Bilirubin*
Published in Karel P. M. Heirwegh, Stanley B. Brown, Bilirubin, 1982
Karel P. M. Heirwegh, Stanley B. Brown
In addition to the often extremely useful structural information imparted by analysis of the azo pigment from the diazo reaction, it has been used for many years in the clinical quantitation of serum bilirubin (van den Bergh reaction)1.22 with particular notice paid to the differing reactivities of bilirubin and its conjugates. Thus, the diazo reaction was placed on a reasonably quantitative basis using visible spectroscopy, and distinction could be made between the faster reacting (“direct-reacting”) conjugated serum bilirubin and the slower (“indirect-reacting”) unconjugated serum bilirubin. Addition of “accelerators”, e.g., ethyl alcohol, to the serum samples causes the slower reacting bilirubin to react considerably faster, probably because its state of aggregation is reduced (see below and Chapter 3).
Diagnostic criteria and contributors to Gilbert’s syndrome
Published in Critical Reviews in Clinical Laboratory Sciences, 2018
Karl-Heinz Wagner, Ryan G. Shiels, Claudia Anna Lang, Nazlisadat Seyed Khoei, Andrew C. Bulmer
Currently, the diazo dye method [96] is the most widely-used assay in clinical laboratories and quantifies the bilirubin content in blood and urine using a diazotized sulfanilic acid solution. The assay detects DBIL (mostly bilirubin glucuronides) or TB (inclusive of unconjugated, albumin bound and bilirubin glucuronides) when solubilizing agents are added. The van den Bergh reaction (diazo test) is widely used because of its low cost, its simplistic design, and its ability to quantify clinically-relevant levels of unconjugated and physiologically conjugated bilirubins [103]. Although the diazo method has these benefits, HPLC analysis is gaining in popularity because it can differentiate between many different bile pigments. These compounds include IBIL and its isomers (IIIα, IXα, and XIIIα), bilirubin mono, or diglucuronide, and biliverdin [81,104]. HPLC analysis is particularly well-suited within the research setting because of its low detection levels. Currently many different HPLC assays for bile pigments have been published [17,86]. More recently, liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods have also been developed [105].