![Calculation: -
Total Bilirubin (mg/dl): - Absorbance of test / Absorbance of standard *Conc. of std.
(mg/dl)
Direct Bilirubin (mg/dl): - Absorbance of test / Absorbance of standard *Conc. of std.
(mg/dl)
[Note: - Total Bilirubin= Indirect Bilirubin- Direct Bilirubin]
Normal value: - Total Bilirubin- 0- 2.0mg/dl. Direct Bilirubin- 0-0.2mg/dl
Clinical Significance: -
Hepatitis, cirrhosis, haemolytic illnesses, a number of inherited enzyme abnormalities,
and obstructive conditions of the bile duct all cause increased total bilirubin.
Pre-hepatic conditions such haemolytic disorders or liver illnesses that limit entrance,
transport, or conjugation inside the liver raise indirect bilirubin levels.
Since indirect (or free) bilirubin coupled to albumin is more likely to pass through the
blood-brain barrier and increase the risk of cerebral injury, monitoring indirect bilirubin
in neonates is especially crucial.](https://image.slidesharecdn.com/serumbilirubintest-250804112321-9ba78f81/85/Liver-Function-Test-Serum-bilirubin-test-pptx-9-320.jpg)
Liver Function Test -Serum bilirubin test.pptx
- 1. Liver Function Test ( Serum Bilirubin Test) PRESENTED BY: SHANTI NATH ASSISTANT PROFESSOR MEWAR UNIVERSITY Department of Paramedical Science
- 4. •Alkaline Phosphatase (ALP): ALP is found in the liver and bile ducts, and its elevation can indicate bile duct obstruction or liver inflammation. •Gamma-Glutamyl Transferase (GGT): GGT is primarily found in the liver and is a sensitive marker for liver damage, especially in the bile ducts. 2. Tests for Bile Flow: •Bilirubin: Bilirubin is a waste product produced during the breakdown of red blood cells. The liver processes bilirubin and excretes it in bile. Elevated bilirubin levels (both direct and indirect) can indicate liver dysfunction, bile duct obstruction, or problems with bilirubin processing. 3. Tests for Overall Liver Function: •Albumin: Albumin is a protein produced by the liver. Low albumin levels can indicate impaired liver function as the liver is unable to produce enough protein. •Total Protein: This test measures the total amount of protein in the blood, including albumin and other proteins. Low total protein can also suggest liver dysfunction. •Prothrombin Time (PT) and International Normalized Ratio (INR):
- 5. BILIRUBIN TOTAL & DIRECT: - Introduction: - After 120 days RBCs is dead and Bilirubin is formed in this process haemoglobin breakdown in heme and globulin. heme again release iron and porphyrin ring. Iron and globulin are returning to the blood circulation for new haemoglobin formation and non- protein part porphyrin is convert into biliverdin. Presence of biliverdin reductase it converts into bilirubin.
- 6. Bilirubin is transported to the liver bound by albumin. This bilirubin is water insoluble and is known as indirect or unconjugated bilirubin. In the liver, bilirubin is conjugated to glucuronic acid to form direct bilirubin this bilirubin is water soluble and also known as conjugated bilirubin. Conjugated bilirubin is excreted via the biliary system into the intestine. Here it is metabolised by bacteria to 2% urobilinogen and 18% stercobilinogen and rest return to the blood.
- 7. Method: - Diazo method. Principle Pearlman & Lee's modified technique, which uses a surfactant as a solubilizer. Direct reaction between bilirubin glucuronate and sulphodiazonium salt produces a coloured derivative. Azobilirubin. The direct bilirubin concentration in the sample is directly correlated with the colour intensity of the produced azobilirubin as measured at 540–550 nm. Reagent composition:-
- 8. REAGENT PREPARATION:- Reagents are liquid, ready to use. Prepare working reagent by mixing of 4 portion of reagent R1 (BIT) or reagent R2 (BID) with 1 portion of reagent R3. STABILITY AND STORAGE:- The unopened reagents are stable till the expiry date stated on the bottle and kit label when stored at 2–8 °C. The working reagent is stable for 7 days at 2–8 °C, when protected from contamination and light. It is recommended to prepare fresh working solution before assay is performed. Stability in serum / plasma: Bilirubin Total: 1 day at 15–25 °C Bilirubin Direct: 2 days at 15–25 °C 6 months at -20 °C 6 months at -20 °C 7 days at 4–8 °C 7 days at 4–8 °C Protect sample from light. Discard contaminated specimens. Procedure: - Take three test tube and label them as T, S, and B. Add 500microlitre working reagent in each tube and mix thoroughly. Add 25 microlitre distilled water in test tube B. Add 25 microlitre standard in test tube S. Mix well, and incubate for 5 minutes at 370 C for total and direct bilirubin. Read absorbance at 546/630 nm against reagent blank.
- 9. Calculation: - Total Bilirubin (mg/dl): - Absorbance of test / Absorbance of standard *Conc. of std. (mg/dl) Direct Bilirubin (mg/dl): - Absorbance of test / Absorbance of standard *Conc. of std. (mg/dl) [Note: - Total Bilirubin= Indirect Bilirubin- Direct Bilirubin] Normal value: - Total Bilirubin- 0- 2.0mg/dl. Direct Bilirubin- 0-0.2mg/dl Clinical Significance: - Hepatitis, cirrhosis, haemolytic illnesses, a number of inherited enzyme abnormalities, and obstructive conditions of the bile duct all cause increased total bilirubin. Pre-hepatic conditions such haemolytic disorders or liver illnesses that limit entrance, transport, or conjugation inside the liver raise indirect bilirubin levels. Since indirect (or free) bilirubin coupled to albumin is more likely to pass through the blood-brain barrier and increase the risk of cerebral injury, monitoring indirect bilirubin in neonates is especially crucial.