T h e C r e a t i v e A p p r o a c h t o B i o s c i e n c e
Urea/BUN - LS
(Modified Urease-Berthlot Method)
REF: 321 001 90 Test REF: 321 002 180 Test
R1 Buffer 1 x 90 ml R1 Buffer 2 x 90 ml
R2 Urease 1 x 1.5 ml R2 Urease 2 x 1.5 ml
R3 Alkaline reagent 1 x 20 ml R3 Alkaline reagent 1 x 40 ml
REF: 321 003 270 Test REF: 321 004 400 Test
R1 Buffer 3 x 90 ml R1 Buffer 4 x 100 ml
R2 Urease 3 x 1.5 ml R2 Urease 4 x 1.7 ml
R3 Alkaline reagent 1 x 65 ml R3 Alkaline reagent 2 x 40 ml
Intended Use Deterioration
Spectrum Diagnostics colorimetric urea reagent is intended for the Do not use the reagent if it is turbid. Failure to recover control values
in-vitro quantitative, diagnostic determination of urea in human serum within the assigned range may be an indication of reagent
on both automated and manual systems. deterioration.
Background Specimen Collection and Preservation
Urea is the major end product of protein nitrogen metabolism. It is Serum
synthesized by the urea cycle in the liver and excreted through the No special preparation of the patient is required. Use nonhaemolyzed
kidneys. The circulating levels of urea depend upon protein intake, serum or plasma [Link] only acceptable anticoagulants are heprin,
protein catabolism and kidney function. Elevated urea levels can EDTA and fluoride. Do not use ammonium heparin plasma.
occur due to renal impairment or in some diseases such as diabetes, Stability: 7 days at 15 –25oC ; 7 days at 2 – 8 oC;
infection, congestive heart failure and during different liver diseases. 1 year at -20 oC
Determination of blood urea nitrogen is the most widely used screening Urine
test for renal function together with serum creatinine. Urine samples are prediluted 1 : 50 with ammonium free water prior
to assay.
Method Stability: 2 days at 15 –25 oC ; 7 days at 2 – 8 oC;
1 month at -20 oC
Urease-colorimetric method.
System Parameters
Assay Principle Wavelength 578 nm (578-623 nm)
Optical path 1 cm
The reaction involved in the assay system is as follows: Assay type End-point
Urea is hydrolyzed in the presence of water and urease to produce Direction increase
ammonia and carbon dioxide. temperature 15-25 oC or 37 oC
Zero adjustment Against Reagent blank
Urea + H2O Urease 2NH3 + CO2 Reagent Blank Limits Low 0.02 AU
High 0.2 AU
The free ammonia in an alkaline pH and in the presence of indicator Sensitivity 0.6 md/dL (0.1 mmol/l)
forms coloured complex proportional to the urea concentration in Linearity 200 mg/dL (33.3 mmol/l)
the specimen.
Procedure
Reagents
Standard urea (ST) Aqueous primary standard
50 mg/dL 8.33 mmol/l Blank Standard Specimen
Reagent 1 (R1 Buffer) Working solution 1.0 ml 1.0 ml 1.0 ml
Phosphate buffer pH 8.0 100 mmol/l
Sodium salicylate 80 mmol/l
Sodium nitroprusside 6.0 mmol/l Standard ----- 10 µl -----
EDTA 30.0 mmol/l Sample ----- ----- 10 µl
Reagent 2 (R2 Enzyme)
Urease >350000 U/l Mix and incubate for at least 3 minutes at 37 oC or 5 minutes at
20-25 oC .
Reagent 3 (R3 Alkaline Reagent)
Sodium hydroxide 400 mmol/l
Sodium hypochlorite 20.0 mmol/l R3(Alk) 200 µl 200 µl 200 µl
Irritant (xi) R36/38: Irritating to eyes and skin. S26: In case of
contact with eyes, rinse immediately with plenty of water and seek Mix and incubate for 5 minutes at 37 oC or 10 minutes at 20-25 oC
medical advice. S37/39: Wear suitable gloves and eye/face protection. Measure absorbance of specimen (Aspecimen) and standard (Astandard)
against reagent blank .
For further information, refer to the Urea/BUN reagent material safety
data sheet.
Calculation
Aspecimen
Precautions and Warnings Serum urea concentration (mg/dl) = xn
Do not ingest or inhalate. In case of contact with eyes or skin; rinse Astandard
immediately with plenty of soap and water. In case of severe injuries; where n = 50.0 mg/dl (8.33 mmol/l)
seek medical advice immediately.
Urine urea concentration is determined by multiplying the result by the
Reagent Preparation , Storage and Stability dilution factor (50).
To prepare the working solution add the content of one vial of
urease (R2) to one bottle of buffer reagent (R1).Once opened, the Urea Nitrogen: To convert the result from urea to urea nitrogen
reagent vial and standard are stable for 3 months at the specified multiply the result by 0.467.
temperature if contamination is avoided.
Stability : 1 Month at 2-8 oC.
Quality Control Anticoagulants
Ammonium heparin should not be used.
Normal & abnormal control serum of known concentrations should
be analyzed with each run. Others
Ammonium ions should be avoided since it may cause erroneously
Performance Characteristics elevated results. Color development in the Berthlot reaction is
Precision suppressed by amines, thiols, steroids and ascorbic acid.
Within run (Repeatiblity)
Expected Values
Level 1 Level 2
Urea(Serum)
n 20 20
Mean (mg/dL) 60 144 Adults <65 years : 15 – 50 mg/dL (2.5-8.33 mmol/L)
Adults >65 years : < 70 mg/dL (<11.66 mmol/L)
SD 1.87 2.1
BUN(Serum)
CV% 3.12 1.46
Adults <65 years : 7 – 23.5 mg/dL
Run to run (Reproducibility) Adults >65 years : 7 – 32.9 mg/dL
Children : 5 – 18 mg/dL
Level 1 Level 2
Urine (24) hours
n 20 20
Mean (mg/dL) 62 146 Urea : 20 – 35 g/24hrs (330-580 mmol/24hrs)
BUN : 9.3 – 16.4 g/24hrs
SD 1.92 2.5
Spectrum Diagnostics does not interpret the results of a clinical
CV% 3.10 1.71 laboratory procedure; interpretation of the results is considered
the responsibility of qualified medical personnel. All indications
Methods Comparison of clinical significance are supported by literature references.
A comparison between Spectrum Diagnostics Urea/BUN reagent Analytical Range
and a commercial reagent of the same methodology was performed
on 20 human sera. A correlation of 0.97 was obtained. 0.6 – 200 mg/dL (0.1 - 33.3 mmol/L).
Sensitivity Waste Disposal
When run as recommended, the minimum detection limit of the This product is made to be used in professional laboratories.
assay is 0.6 mg/dL. Please consult local regulations for a correct waste disposal.
S56: dispose of this material and its container at hazardous or
Linearity special waste collection point.
S57: use appropriate container to avoid environmental contamination.
The reaction is linear up to a urea concentration of (200 mg/dl) S61: avoid release in environment. refer to special instructions/safety
33.3 mmol/L. Specimens showing higher concentrations should data sheets.
be diluted 1+2 with physiological saline and repeat the assay
(result×3). References
Interfering Substances 1. Batton, C. J & crouch, S.R : Anal. Chem., 1977,49:464-469.
Serum, plasma 2. Shephard MD, Mezzachi RD : Clin Biochem Revs, 4:61-7, 1983.
3. Tietz NW, ED. Clinical guide to Laboratory tests. 2ND ED.
Haemolysis Philadelphia: WB Saunders; 1990:566.
Erythrocyte contamination doesn’t elevate results. 4. Tiffany to, jansen JM, Burtis CA,Overton JB, Scott CD. Enzymatic
Kinetic Rate and end Point analyses of Substrate, By USE of A
Icterus Gemsaec fast analyzer. Clin Chem.
No significant interference.
Lipemia
Lipemic specimens interfere with the method of Berthlot. ORDERING INFORMATION
CATALOG NO. QUANTITY
321 001 1 x 90 ml
321 002 2 x 90 ml
321 003 3 x 90 ml
321 004 4 x 100 ml
Egyptian Company for Biotechnology (S.A.E)
Obour city industrial area. block 20008 piece 19 A. Cairo. Egypt.
Tel: +202 4489 2248 - Fax: +202 4489 2247
[Link]
E-mail:info@[Link]
MDSS GmbH
EC REP Schiffgraben 41
30175 Hannover, Germany
IFUFCC47 Rev.(2), 9/6/2022