2 Objective The aim of the experiment is to determine : Total bilirubin Direct bilirubinIndirect bilirubin , Contents of human serum .
3 IntroductionThe liver responsible for many important metabolic functions> 100 lab tests associated with liver functionMajor hepatic functionsProtein synthesisRegulation of carbohydrate metabolismRegulation of lipoproteinsDetoxification of drugs and chemicalsExcretion of bileFormation of bilirubin
4 Introduction Alkaline phosphatase SGOT ( AST) SGPT ( ALT) These enzymes are released from dameged liver cells
5 Extravascular Pathway for RBC Destruction (Liver, Bone marrow,& Spleen)Phagocytosis & LysisHemoglobinGlobinHemeBilirubinAmino acidsFe2+Amino acid poolRecycledExcreted
6 DEGRADATION OF HEME TO BILIRUBIN Bilirubin is the main pigment derived from heme catabolism75% is derived from RBCsIn normal adults this results in a daily load of 200 mg of bilirubinNormal plasma concentrations are less then 1 mg/dLHydrophobic – transported by albumin to the liver for further metabolism prior to its excretionP450 cytochrome“unconjugated” bilirubin
8 HYPERBILIRUBINEMIA-- elevated bilirubin in serum (above 3mg/dL) occur when there is imbalance between production and excretion-- can be conjugated or unconjugated or both depending on the situation-- elevated bilirubin can diffuse into tissues, making them appear yellow (jaundice)
9 Clinical Consequences: HYPERBILIRUBINEMIAClinical Consequences:-- Conjugated hyperbilirubinemia: benign-- Unconjugated hyperbilirubinemia: benign at concentrations < 25 mg/dL (albumin capacity)-- At concentrations >25 mg/dL, unconjugated bilirubin is free (uncomplexed) and can enter the brain.bilirubin encephalopathy (kernicterus)
10 Causes of JAUNDICE( per-hepatic_ the pathology is occurring prior to the liver ) Ex. Hemolytic anemia-- destruction of erythrocytes( hepatic _ the pathologyis located within the liver ) a)Hepatitis b) cirrhosis-- conjugation and excretion of bilirubinc) Neonatal “physiological jaundice”-- immature hepatic system of the newborn: uptake, conjugation, excretion of bilirubin3) ( post-hepatic _ _ the pathology is occurring poster to the liver ) Bile duct obstruction-- conjugated bilirubin not delivered to intestine;it backs up, spills over into the blood
11 principle Direct bilirubin water soluble Bilirubin + Diazo reagent Red –blue color* Only direct bilirubin react .
12 principle Direct bilirubin water soluble Bilirubin + Diazo reagent + caffeineRed –blue colorDirect + indirect bilirubin react .
13 Total bilirubin = Conjugated + Un conjugated Bilirubin.