TOTAL BILIRUBIN REAGENT
INTENDED USE
Total Bilirubin Reagent can be used for the quantitative determination of total bilirubin in human serum.
INTRODUCTION
Bilirubin is a metabolite of the heme portion of heme proteins, mainly hemoglobin. Bilirubin is excreted into the intestine and bile from the liver. The site of the catabolism of hemoglobin is the reticuloendothelial system (RES). Bilirubin is then released into the bloodstream where it binds tightly to albumin and is transported to the liver. Upon uptake by the liver, bilirubin is conjugated with glucuronic acid to form bilirubin mono and diglucuronide, which are
water-soluble metabolites. The metabolites are then excreted in the
bile.
Elevation of total serum bilirubin may occur due to (1) excessive hemolysis or destruction of the red blood cells (e.g. haemolytic disease of the newborn), (2) liver diseases (e.g. hepatitis and cirrhosis), (3) obstruction of the biliary tract (e.g.. gallstones). Most chemical methods for the determination of total bilirubin are based on the reaction between diazotized sulfanilic acid and bilirubin to produce azobilirubin, which absorbs maximally at 560 nm. Such tests are often run in the presence or absence of an organic solvent e.g., methanol to distinguish free bilirubin from conjugated bilirubin on a differential solubility basis.
PRINCIPLE
Total Bilirubin Reagent
Bilirubin reacts with diazotized sulfanilic acid to produce azobilirubin, which has an absorbance maximum at 560 nm in the dimethyl sulfoxide (DMSO) solvent. The intensity of the color produced is directly proportional to the amount of total bilirubin concentration present in the sample.
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