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Cls331,,,experiment (8):. Determination of serum urea. Introduction:. Urea or carbamide is an organic compound with the chemical formula ( N H 2 ) 2 C O . The molecule has two amine (-NH 2 ) groups joined by a carbonyl (C=O) functional group .

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cls331 experiment 8

Cls331,,,experiment (8):

Determination of serum urea

introduction
Introduction:

Urea or carbamide is an organic compound with the chemical formula (NH2)2CO. The molecule has two amine (-NH2) groups joined by a carbonyl (C=O) functional group.

slide3

Urea is the main end product of protein metabolism in the body, which takes place in the liver.

  • Normally cleared from the blood by the kidney into the urine. Diseases that compromise the function of the kidney often lead to increased blood levels of urea, as measured by the blood urea nitrogen (BUN) test.
principle
Principle:
  • Urea is hydrolized by the enzyme urease forming ammonia and carbon dioxide.
  • NH2CO NH2 + H2O urease
  • NH3 +NH2 CO OH 2NH3 +CO2
  • NOTES:
  • Urease acts optimally at 55⁰c & PH (7-8)
  • Inhibited by ammonia & fluoride.
o phthalaldehyde colorimetric method
o-Phthalaldehyde colorimetric method
  • The principle:

Urea + o-fhthalalehyde Isoindoline

Clinical significance:

ureain blood(uremia): diets with excess of proteins, renal diseases, renal obstruction and heart failure…

slide7

Mix and incubate 15 min. at 37⁰c.

  • Read the (A) at 510 nm.
  • Calculations;
  • Sample × 50 = mg/dl

(A) std.

Reference values: 15 – 45 mg/dl