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Estimation of Urea by DAM Method

This document describes the diacetyl monoxime method (DAM method) for estimating blood urea. Urea reacts with diacetyl monoxime under acidic conditions in the presence of ferric ions and thiosemicarbazide to form a pink colored complex. The intensity of the color corresponds to the amount of urea present and is measured using a green filter at 540nm. The procedure involves preparing standards and samples, adding the color and acid reagents, incubating in a boiling water bath, cooling, and measuring absorbance. The urea concentration in a blood sample is calculated using the standard absorbance and a factor to convert to mg/dl. Normal blood urea levels are 15-45 mg

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3K views3 pages

Estimation of Urea by DAM Method

This document describes the diacetyl monoxime method (DAM method) for estimating blood urea. Urea reacts with diacetyl monoxime under acidic conditions in the presence of ferric ions and thiosemicarbazide to form a pink colored complex. The intensity of the color corresponds to the amount of urea present and is measured using a green filter at 540nm. The procedure involves preparing standards and samples, adding the color and acid reagents, incubating in a boiling water bath, cooling, and measuring absorbance. The urea concentration in a blood sample is calculated using the standard absorbance and a factor to convert to mg/dl. Normal blood urea levels are 15-45 mg

Uploaded by

mdanasd123
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© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Estimation of Blood Urea by DAM

(Diacetyl Monoxime Method)

Aim: To estimate blood Urea by DAM method


Method: Diacetyl monoxime method (DAM method)
Principle: Urea reacts with Diacetyl monoxime under strong acidic
condition in presence of ferric ions and thiosemicarbazide to give a pink
colored complex. Intensity of color is a measure of amount of urea
present in blood. Color intensity is compared with standard and is
measured using green filter (540nm).

Reagents:
1. Preparation of standard: Take 1mg of urea and add 100ml of
distilled water.
2. Composition of mixed colour reagent: Diacetly monoxime and
Thiosemicarbazide
3. Composition of mixed acid reagent: Orthophosphoric acid
concentrated H2SO4 , Ferric chloride.
4. Diluted sample: 1:100

Procedure: Label three test tubes as B (Blank), T (Test), and S


(Standard). Pipette 1mL water into B, 1mL of diluted serum (1:100) into
T and 1mL of standard urea solution ( 1mg in 100ml of distilled water )
into S. Add 2mL of diacetyl monoxime(DAM) solution and 2mL of acid
reagent into each tube. Mix well and keep in a boiling water bath for 20
minutes. Mix well and keep in a boiling water bath for 20minutes.cool at
temperature and take OD at 540nm.

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Tabular column:

Reagents Blank Standard Test

Distilled water 1mL - -


Standard - 1mL -
Diluted Serum - - 1mL
Mixed colour reagent(mL) 2 2 2
Mixed acid reagent (mL) 2 2 2
Mix well and keep in the boiling waterbath for 12minutes and then cool under tap
water
Absorbance at 540nm

Calculation:
Concentration of urea = ODT – ODB Χ CS Χ 100
in mg/100dl of blood ODS – ODB VS
= ODT – ODB Χ 0.01 Χ 100
ODS – ODB 0.01

= ....................... mg/dl
Blood urea Nitrogen = Blood urea level
2.14
Report: The amount of urea present in the given blood sample is.....mg/dl
Clinical significance:

 Normal values: Normal blood urea = 15-45 mg/dl, slightly higher


in males
 Normal BUN: 10-20mg/dl

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Causes of increased blood urea level:
1. Pre-renal causes: Since blood supply to the kidney is reduced,
filtrated and excreted of urea is minimum
a. High protein diet
b. Increases with age vomiting
c. Dehydration as in diarrhea, vomiting
d. Increased cardiac output/ failure
e. Increased catabolism of proteins as in fever and wasting diseases.
2. Renal causes: Synthesis of urea is normal. But damage in the renal
tissue leads to poor filtration and excretion of urea.
a. Nephritis
b. Nephrotic syndrome
c. Acute renal failure
d. Chronic renal failure
e. Polycystic kidney
f. Hydronephrosis
g. Malignant hypertension
3. Postreanal causes: synthesis and filtration are normal but renal
passage is blocked. Hence minimum excretion of urea occurs.
a. Obstruction in the renal tract
b. Enlarged of prostate
c. Stones in bladder
 Blood urea level decreases in liver diseases due to decreased
synthesis
 Blood urea level is commonly monitored to evaluate kidney
diseases.
 A high urea concentration in the urine reflects the concentrating
power of the kidney
 Relationship between blood urea nitrogen(BUN) and urea is blood
urea nitrogen(mg/dl) = urea (mg/dl)/2.14
 Urea level is generally studied in conjuction with creatnine level to
identify renal dysfunction. Urea/creatinine ratio is sometimes used
to discriminate between prerenal ansd postrenal uremia.
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