BUN: KINETIC QUANTITATIVE DETERMINATION OF - aqueous solution of urea with traces of

UREA NITROGEN (BUN) IN SERUM amounts of stabilizers

SUMMARY AND PRINCIPLE:
 Urea is the principle waste product of protein
catabolism
 synthesized in the liver from ammonia,
produced as a result of the deamination of
amino acids
 comprises only about 45% of the non-protein
nitrogen
 its values is important in determining liver and
kidney functions
Decrease in BUN: nephritis, acute liver destruction,
amyloidosis, pregnancy
Increase in BUN: acute and chronic nephritis, intestinal
and urinary obstruction, uremia, metallic poisoning,
pneumonia, Addison’s disease, peritonitis, surgical
shock, cardiac failure
- Stanbio BUN procedure is a modification of the
method described by Sampson
1. Urea is catalytically converted to ammonium
carbonate with: Urease
2. Rate of reaction is dependent upon
concentration of: glutamic dehydrogenase
3. Rate of this second reaction depends upon the
first and can be measured by rate of conversion
of NADH to NAD by the change of absorbance at
340 nm
- reference against NIST material
PRECAUTION: FOR IN VITRO DIAGNOSTIC USE
- reagent contains sodium azide,
toxic if ingested
- may also react with lead and copper
plumbing, forming highly explosive
metal azides
REAGENT PREPARATION:
1. Buffer and enzyme liquid reagents are supplied
read-to-use
2. Prepare working reagent. 5 parts Buffer to 1
part Enzyme
3. Before use, allow to stand at least 30 minutes at
room temperature (15-25 C)
REAGENT STORAGE AND STABILITY:
- store at 2-8 degrees Celsius
- protected from light
- should appear clear and colorless
- discard if it appears cloudy or has
particulate matter
 Working reagent – stable for 4 weeks (2-8 C) or
5 days (15-25 C)
- discarded if initial absorbance read
against distilled water at 340 nm is
below 1.000
SPECIMEN COLLECTION AND PREPARATION:

REAGENTS: - Non-hemolyzed serum is the

specimen of choice
BUN Buffer Ref. No. 2021
- should be separated and analyzed
 composition: on the day of collection
- Tris. pH 7.8 120 mmol/L - avoid anticoagulants containing
- 2-Oxoglutarate 7.0 mmol/L ammonium or fluoride salts
- ADP (monosodium salt) 0.6 mmol/L  Urea in serum: stable for 24 hours (room
- Urease (Jack Bean) 60000 U/L temperature 15-25 C)
- GLDH (Backer’s yeast) 10000 U/L - several days (2-8 C)
- 2-3 months (-20 C)
BUN Enzyme
INTERFERING SUBSTANCES: hemolysis, fluoride at
 composition: elevated concentrations, ammonia
- NADH 0.25 mmol/L

BUN Standard – 30mg/dL

RESULTS: values are derived by:

- comparing the absorbance change

of unknown with that of a standard
 Serum BUN (mg/dL) = Au/As x 30

LIMITATIONS: If BUN values exceeds 140 mg/dL,

specimen should be diluted 2 fold with distilled water

EXPECTED VALUES: