Bilirubin is a yellow breakdown product of heme catabolism that is excreted in bile and urine. Elevated bilirubin levels can indicate liver or blood disorders. Bilirubin exists in conjugated and unconjugated forms, and levels of each are measured to determine the type of jaundice. Differential diagnoses of jaundice include pre-hepatic causes that increase unconjugated bilirubin or post-hepatic causes that increase both conjugated and unconjugated bilirubin. Bilirubin is measured using direct spectral methods or diazo reactions to determine the type and cause of jaundice.
Bilirubin is a yellow breakdown product of heme catabolism that is excreted in bile and urine. Elevated bilirubin levels can indicate liver or blood disorders. Bilirubin exists in conjugated and unconjugated forms, and levels of each are measured to determine the type of jaundice. Differential diagnoses of jaundice include pre-hepatic causes that increase unconjugated bilirubin or post-hepatic causes that increase both conjugated and unconjugated bilirubin. Bilirubin is measured using direct spectral methods or diazo reactions to determine the type and cause of jaundice.
Bilirubin is a yellow breakdown product of heme catabolism that is excreted in bile and urine. Elevated bilirubin levels can indicate liver or blood disorders. Bilirubin exists in conjugated and unconjugated forms, and levels of each are measured to determine the type of jaundice. Differential diagnoses of jaundice include pre-hepatic causes that increase unconjugated bilirubin or post-hepatic causes that increase both conjugated and unconjugated bilirubin. Bilirubin is measured using direct spectral methods or diazo reactions to determine the type and cause of jaundice.
Bilirubin is a yellow breakdown product of heme catabolism that is excreted in bile and urine. Elevated bilirubin levels can indicate liver or blood disorders. Bilirubin exists in conjugated and unconjugated forms, and levels of each are measured to determine the type of jaundice. Differential diagnoses of jaundice include pre-hepatic causes that increase unconjugated bilirubin or post-hepatic causes that increase both conjugated and unconjugated bilirubin. Bilirubin is measured using direct spectral methods or diazo reactions to determine the type and cause of jaundice.
Clinically hyperbilirubinemia appears as jaundice or
Yellow breakdown product of normal heme icterus catabolism Jaundice can be usually detected when the serum Excreted in the bile and urine (elevated levels may bilirubin level exceeds 2.0 - 2.5 mg/dL indicate certain diseases) When the level of bilirubin is between 1-2 mg/dL, it Responsible for the yellow color of bruises, the is known as Latent Jaundice background straw-yellow color of urine In neonates unconjugated bilirubin can cross BBB The brown color of feces (via its conversion to (blood brain barrier) and thus lead to accumulation in stercobilin), and yellow discoloration in jaundice brain of neonates. This interfere with nervous system End product of hemoglobin and serve as a diagnostic development and permanent nervous impairment marker of liver and blood disorders
FUNCTIONS DIFFERENCE BETWEEN CONJUGATED AND
Levels of serum bilirubin are inversely related to risk UNCONJUGATED BILIRUBIN of certain heart disease CONJUGATED UNCONJUGATED Present normally in bile Present normally in plasma Acts as uncoupler in neonates and thus maintain body Conjugated to glucoronic acid Conjugated to albumin heat Can be filtered by the kidney Cannot be filtered by the Bile pigments such as biliverdin naturally possess kidney significant anti-mutagenic and antioxidant properties Cannot cross BBB Can cross BBB Biliverdin and bilirubin have shown to be potent scavengers of peroxyl radicals Inhibit the effcts of polycyclic aromatic hydocarbons, IN THE LABORATORY, BLIRUBIN IS MEASURED AS: heterocylci amines, and antioxidants – all of which are 1. Total bilirubin mutagens 2. Indirect or Unconjugated bilirubin 3. Direct or conjugated bilirubin FORMS OF BILIRUBIN Unconjugated Conjugated Structure Free bilirubin; B1 Bilirubin CHANGES IN CONCENTRATION OF BILIRUBIN diglucoronide; B2 IN THOSE WITH JAUNDICE Other name Hemobilirubin; Choleglobin; TOTAL CONJUGATED Hemolytic bilirubin; Cholebilirubin; One- TYPE OF JAUNDICE BILIRUBIN BILIRUBIN UNCONJUGATED
Prehepatic bilirubin minute bilirubin; PRE HEPATIC
Prompt bilirubin; HDN Normal Cholestatic bilirubin; Physiologic Normal Post-hepatic Diseases of the Types of Non-polar Polar Newborn Compound Ineffective Increase Normal Increase Solubility Hematopoesis Water Insoluble Soluble Crigler-Najar Decrease Alcohol Soluble Soluble Syndrome Van Den High affinity; Indirect Low affinity; Direct Gilberts Normal Berg Syndrome reaction HEPATIC Urine: Absent Present Viral Increase Increase Increase Hemolytic Dubin Johnson Normal Urine: Rotor syndorme Normal Obstructive POST HEPATIC Positive (+) Negative (–) Increase Increase Increase Affinity to OBSTRUCTION brain tissue
FRACTIONS OF BILIRUBIN Indirect Blilirubin = Total Bilirubin – Direct Bilirubin
Alpha – unconjugated Beta – mono conjugated Gamma – diconjugated Delta – irreversibly bound to albumin. Usually seen in solutions with problems in the extinction of conjugated bilirubin Note: Alpha beta and gamma have a directly out with diazo reagent DIFFERENTIAL DIAGNOSIS BILIRUBIN DETERMINATION DIRECT SPECTRAL METHOD DECREASED ICTERUS INDEX OF MUELENGRACHT TRANSPORT - It is based on the degree of icterus in the serum or INCREASED OR IMPAIRED BILIARY PRODUCTION DECREASED EXCRETION OBSTURCTION plasma. As compared to the standard potassium CONJUGATION dichromate solution normal value is 2-6 icterus Increased UB Increased CB index units. Hemolysis Gilbert’s Rotor’s CB/CBH Syndrome Disease Syndrome - Not reliable – pigments (carotene; xanthophyll) Transfusion Crigler Najjar Dubin- Stricture JACKSON DIRECT BILIRUBINOMETER Johnson - The serum is diluted with 15 m phosphate buffer Transfusion Neonatal Cancer Tumor/ and the absorbance is read at 450 mm and 575 nm Reaction Cancer using a narrow band filter. Standard methyl Sepsis Cirrhosis Cirrhosis Pancreatitis orange at pH of 7.4 Burns Hepatitis Hepatitis Primary Note: it is very rapid and usually use among Sclerotizing infants but not in adults due to interferences Cholangitis - Bilirubin (mg/dl) = A450 - A575 x 50 Hemoglobino Drug Amyloidosis Parasitic pathies Inhibition Infections DIAZOTIZATION PRINCIPLE by Van den Berg Reaction Pregnancy – based on the reaction of bilirubin with diazotized sulfanilic acid producing a colored pigment azobilirubin (red color) FUNCTION PRE POST Evelyn-Malloy method HEPATIC TEST HEPATIC HEPATIC Jendrassik-Grof method Total Bilirubin Normal/ Increased Increased Rand and Di Pasqua method Increase Michaelson Conjugated Normal Increased Increased Gindler-Ishizaki Unconjugated Normal/ Increased Normal Anino Watson and Ducci Increase Stoner and Wisberg Urobilinogen Normal/ Increased Decreased/ Thamhausser Anderson Increase Negative Fluorometry Urine Color Dark Dark Dark (Urobilinogen) (Urobilinogen (Conjugated Continuous Flow Technique by Gambino + Conjugated Bilirubin) Bilirubin) Basis Evelyn- Jendrassik- Rand & Di Stool Color Normal Normal Pale Malloy Grof Pasqua Alkaline Product Red Blue Red Phosphatase Normal Increased Increased Measured Violet Azobilirubin Azobilirubin Level Azobilirubin Alanine Incubation Db: 1 minute Db: 5 minutes 5 minutes Transferase Period Tb: 15 minutes Tb: 10 minutes and Aspartate Normal Increased Increased pH Acidic to Basic Transferase Neutral Levels Accelerator 50% Methanol Caffeine Conjugated Sodium Bilirubin in Absent Present Present Benzoate Urine (Coupling Promoter) Buffer None Sodium SPECIMEN CONSDERATION Acetate Other Diazo Reagent Diazo Reagent Dicholor Light sensitive analyte Reagent and Blank; Benzene Cover with foil or carbon paper both collection tubes Ascorbic Acid Diazonoum and plasma or serum container after separation of use and Alkaline Chloride of specialized tube which is amber colored Tartarate Prevent life policies and lipemia Diazo Reagent: 1.0% sulfanilic acid + 0.5% sodium nitrite SERUM BILIRUBIN Diazo Blank: 1.5% HCL Hyperbilirubinemia – plasma bilirubin exceeds 1 Ascorbic Acid – used to determine diazo reaction mg/dL Alkaline Tartrate – shift peak absorbance of blilirubin from 586 to 600 nm (converts pink to blue) Latent jaundice – between 1 to 2mg/dL >2m g/dL – yellowish discoloration sclera Advantages of Jendrassik-Groff conjunctiva skin mucous membrane resulting in Insensitive to pH change end 50 fold variation in change of jaundice the concentration of the sample Jaundice or Icterus – a condition characterized by Adequate optical sensitivity (low bilirubin); minimal yellowish discoloration of the skin sclera and turbidity and relatively constant blank mucous membrane Not affected by hemoglobin up to 750 mg/dL