Estimation of Serum Bilirubin: Jendrassik-Grof Method

Introduction

Serum bilirubin estimation is crucial for diagnosing liver function disorders, hemolytic anemia, and jaundice. The Jendrassik-Grof method is a widely used colorimetric technique.

Principle

The Jendrassik-Grof method is based on the reaction of bilirubin with diazotized sulfanilic acid, forming a colored complex. This reaction is stabilized using caffeine, allowing accurate measurement using a spectrophotometer.

Procedure

• Sample: Collect venous blood and separate serum.
• Reagents: Diazotized sulfanilic acid, caffeine reagent, acidic solution.
• Reaction: Mix serum with reagents, incubate, and measure absorbance at 546 nm.
• Calculation: Compare absorbance against a bilirubin standard curve.

Clinical Significance

Elevated bilirubin levels indicate liver disease, bile duct obstruction, or hemolysis. Reference values:

• Normal total bilirubin: 0.3 - 1.2 mg/dL
• Direct bilirubin: <0.3 mg/dL

Advantages of Jendrassik-Grof Method

• High specificity and sensitivity.
• Minimized interference from hemoglobin and other substances.
• Reliable and reproducible for clinical diagnosis.

Conclusion

The Jendrassik-Grof method remains a gold standard for bilirubin estimation, offering precision and accuracy crucial for hepatic disorder assessment.