Veterinary Clinical Biochemistry and Laboratory Diagnosis

ESTIMATION OF UREA AND BUN - PROCEDURE

Specimen

• Serum/Plasma

Reagents :[reagents in the kit vary from manufacturer to

                   manufacturer   so the procedure in  the kit used may be followed.]

• Urea - Working standard
• Urea Reagent
• Diacetylmonoxime (DAM)

Preparation of working solution

• Solution I: Dilute one ml of Urea Reagent (1) to 5ml with distilled water.

Procedure (Agappe Diagnostics Kit)

• Mark three test tubes as Blank, Standard and Test and proceed as follows

S.No.	Reagents	Blank (B)	Standard (S)	Test(T)
1.	Solution I	2.5 ml	2.5 ml	2.5 ml
2.	Working Urea Standard (30 mg%)	-	10 µl	-
3.	Serum / Plasma	-	-	10 µl
Mix Well
4.	Diacetylmonoxime (DAM)	250 µl	250 µl	250 µl

• Mix well and keep the tubes in boiling water, exactly for 10 minutes. Cool them immediately under running water for 5 minutes and mix by inversion.
  Measure the optical density within 10 minutes at 525 nm or using a green filter.
  

Calculations

Result

• The amount of urea in the given serum = -------mg/dl
• The amount of BUN in the given serum = -------mg/dl