Red blood cells which contain hemoglobin are constantly being hemolysed, resulting in the hemoglobin they carry

to be degraded to give globin and heme and iron(Ophart,2003). This disintegration process takes place mostly in the livers,spleens and bone marrows reticuloendothelial cells(Ophart,2003). The heme product breaks apart to give biliverdin which is a green coloured pigment(Ophart,2003). This then undergoes rapid reduction to form bilirubin ,an orange coloured pigment(Ophart,2003).

Society of Chemisty,2014).

Figure 1: Structure of bilirubin C33H36N4O6(Royal

The structure of the heme molecule is a ring of four pyrroles connected via carbon bridges surrounding a central iron atom(Orfei,2014). This molecule undergoes sequential catalytic degradation via the use of the enzymes: heme oxygenase and biliverdiin reductase(Orfei,2014). An oxygenase breaks the ring at the alpha bridge to produce a tetrapyrrolic chain with no iron. This is the unconjugated bilirubin (Orfei,2014) .It is in a lipid soluble form(RnCeus.com,1999).Being a non-polar pigment which gives an indirect reaction with alcohol(RnCeus.com,1999).Heme accounts for 85% of daily bilirubin production(Richardson,2008). As the uncongugated bilirubin is toxic to tissues, it is bound to albumin only allowing for its free form to exist ~1 minute(Orfei,2014).It is then transported in the blood to the liver(Orfei,2014). Here hepatocytes take it up on their sinusoidal suface(Orfei,2014).There, the bond between the bilirubin and albumin is broken and the albumin remains in the plasma(Orfei,2014).The bilirubin is rapidly uptaken by the hepatocyte(Orfei,2014) .Here the toxic product is bound to ligandins and Z protein which are cytoplasmic proteins(Orfei,2014).These proteins ensure that a reflux of bilirubin in to the blood is prevented(Orfei,2014). In order to neutralize this toxic product it is conjugated with glycosyl,a modified sugar(Orfei,2014).Here glucuronic acid is conjugated to the bilirubin making it hydrophilic and polar(Orfei,2014).Now can be eliminated in the bile(Orfei,2014). Glucuronic acid (GA) is synthesized from cytosolic glucose in the presence of UP-glucose dehydrogenate(Orfei,2014).This then complexes with uridinediphophate(UDP) ,in the

presences of bilirubin-DUgAN-transferees to produce udpglucuronic acid(UDPGA) (Orfei,2014).The glucuronic acid from UDPGA is then transferred to the bilirubin(Orfei,2014). This entire conjugation process occurs in the endoplasmic reticulum(Orfei,2014). The entire reaction results in an ester being formed between glucuronic acid and either 1 or 2 propionic side-chains of the bilirubin(Orfei,2014).There are bilirubin mono and diglucuronides which are formed in a ratio of 80% to 20% respectively(Orfei,2014). The conjugated form is then secreted into the bile and enters the duodenum(Richardson,2008).It is then reduced to urobilins(Richardson,2008). The cascade is as follows: Bilirubin glucuronide + bacterial or intestinal beta-glucuronidase= free bilirubin(Orfei,2014) Free bilirubin + bacterial dehydrogenase = urobilinogen(colourless) (Orfei,2014) Urobilinogen + dehydrogenase = urobilin (orange-yellow) (Orfei,2014) The majority of the urobilinogen,urobilin and bilirubin is excreted in the feces(Orfei,2014). Some urobilinogen is absorbed back into the blood and then transported to the liver for re-excretion into the bile (Ophart,2003) .It can also enter the blood and then into the kidneys and then be excreted in urine(Ophart,2003). Urine only contains conjugated bilirubin upon exceeding normal levels in the plasma(Orfei,2014).It is not a component in the urine of normal persons a small fraction of non-protein bound bilirubin from the plasma is passed in the urine(Orfei,2014).

Figure 2: Mechanism for production of bilirubin form heme (Photonics4Life,2010). Lab test are able to distinguish between conjugated and unconjugated bilirubin due to their chemical difference ,that is their non-polar and polar nature respectively (RnCeus.com,1999) . The former reacts directly upon addition of dyes as it is water soluble,

while the latter requires the presence of alcohol to react with dyes, making it an indirect measurement(RnCeus.com,1999). As dicussed above, the liver is responsible for converting the toxic substance into a soluble form to be excreted. Abnormal bilirubin levels would indicate that there is some malfunction in the liver or surrounding delivery routes form the liver. Bilirubin testing is done mainly to check liver function and monitor it for signs of liver disease including hepatitis and cirrhosis(WebMD,2014).It can also be used to determine if medications are damaging the liver(WebMD,2014). Blocks in the bile duct, such as gallstones and pancreatic tumors among others can also be detected via the test(WebMD,2014). Conditions that result in the increased destruction of red blood cell including hemolytic anemia and hemolytic disease of newborns can be diagnosed form the results of the test(WebMD,2014). Finally, it is integral in deciding on whether or not new born babies should undergo treatment(WebMD,2014).