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Urea/BUN - Liquizyme SYMBOLS IN PRODUCT LABELLING
(Modified Urease-Berthlot Method) EC REP Authorised Representative Use by/Expiration Date
IVD For in-vitro diagnostic use CAUTION. Consult instructions
REF: 318 001 100 test REF: 318 002 200 test LOT Batch Code/Lot number for use
REF Catalogue Number Manufactured by
R1 Buffer 1 x 100 ml R1 Buffer 2 x 100 ml
R2 Urease 1 x 6 ml R2 Urease 2 x 6 ml o
Consult instructions for use (Xi) - Irritant
C
R3 Alkaline reagent 1 x 20 ml R3 Alkaline reagent 1 x 45 ml o
C Temperature Limitation
REF: 318 003 500 test REF: 318 004 1000 test
R1 Buffer 5 x 100 ml R1 Buffer 4 x 250 ml
R2 Urease 2 x 15 ml R2 Urease 51 ml
R3 Alkaline reagent 2 x 55 ml R3 Alkaline reagent 1 x 210 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 non haemolyzed
kidneys. The circulating levels of urea depend upon protein intake, serum or plasma [Link] only acceptable anticoagulants are heparin,
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 mg/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 1
Reagents
Standard urea (ST) Aqueous primary standard Blank Standard Specimen
50 mg/dL 8.33 mmol/l
R1(Buffer) 1.0 ml 1.0 ml 1.0 ml
Reagent 1 (R1 Buffer)
Phosphate buffer pH 8.0 100 mmol/l R2(Enzyme) one drop one drop one drop
Sodium salicylate 80 mmol/l (50 µl) (50 µl) (50 µl)
Sodium nitroprusside 6.0 mmol/l
EDTA 30.0 mmol/l Standard ----- 10 µl -----
Sample ----- ----- 10 µl
Reagent 2 (R2 Enzyme)
Urease >6000 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 R3([Link]) 200 µl 200 µl 200 µl
Sodium hypochlorite 20.0 mmol/l
Irritant (xi) R36/38: Irritating to eyes and skin. S26: In case of Mix and incubate for 5 minutes at 37 oC or 10 minutes at 20-25 oC
contact with eyes, rinse immediately with plenty of water and seek Measure absorbance of specimen ( A specimen ) and standard
medical advice. S37/39: Wear suitable gloves and eye/face protection. (Astandard) against reagent blank.
For further information, refer to the Urea/BUN reagent material safety Procedure 2 (Using working solution)
data sheet.
Blank Standard Specimen
Precautions and Warnings
Do not ingest or inhalate. In case of contact with eyes or skin; rinse Working solution 1.0 ml 1.0 ml 1.0 ml
immediately with plenty of soap and water. In case of severe injuries; Standard ----- 10 µl -----
seek medical advice immediately. Sample ----- ----- 10 µl
Reagent Preparation, Storage and Stability Mix and incubate for at least 3 minutes at 37 oC or 5 minutes at
Spectrum colorimetric urea reagents are supplied ready-to-use and 20 -25 oC.
stable up to the expiry date labeled on the bottles (2 – 8 oC).
NB: For mega labs having high numbers of patient specimens,working R3([Link]) 200 µl 200 µl 200 µl
buffer reagent can be prepared .( Stability 1 week )
Mix and incubate for 5 minutes at 37 oC or 10 minutes at 20-25 oC.
REF:318 001: add 5 ml from R2 to one bottle of R1; mix gently. Measure absorbance of specimen (Aspecimen) and standard
REF:318 002: add 5 ml from R2 to one bottle of R1; mix gently. (Astandard) against reagent blank.
REF:318 003: add 5 ml from R2 to one bottle of R1; mix gently.
REF:318 004: add 12.5 ml from R2 to one bottle of R1; mix gently.
Calculation Anticoagulants
Aspecimen Ammonium heparin should not be used.
Serum urea concentration (mg/dl) = xn
Astandard Others
where n = 50.0 mg/dl (8.33 mmol/l) Ammonium ions should be avoided since it may cause erroneously
elevated results. Color development in the Berthlot reaction is
Urine urea concentration is determined by multiplying the result by suppressed by amines, thiols, steroids and ascorbic acid.
the dilution factor (50).
Expected Values
Urea Nitrogen: To convert the result from urea to urea nitrogen
multiply the result by 0.467. Urea(Serum)
Quality Control Adults <65 years : 15 – 50 mg/dL (2.5-8.33 mmol/L)
Adults >65 years : < 70 mg/dL (<11.66 mmol/L)
Normal & abnormal control serum of known concentrations should
be analyzed with each run. BUN(Serum)
Performance Characteristics Adults <65 years : 7 – 23.5 mg/dL
Precision Adults >65 years : 7 – 32.9 mg/dL
Within run (Repeatability) Children : 5 – 18 mg/dL
Level 1 Level 2 Urine (24) hours
n 20 20 Urea : 20 – 35 g/24hrs (330-580 mmol/24hrs)
BUN : 9.3 – 16.4 g/24hrs
Mean (mg/dL) 60 144
SD 1.87 2.1 Spectrum Diagnostics does not interpret the results of a clinical
laboratory procedure; interpretation of the results is considered
CV% 3.12 1.46 the responsibility of qualified medical personnel. All indications
of clinical significance are supported by literature references.
Run to run (Reproducibility)
Analytical Range
Level 1 Level 2
n 20 20 0.6 – 200 mg/dL (0.1 - 33.3 mmol/L).
Mean (mg/dL) 62 146 Waste Disposal
SD 1.92 2.5
This product is made to be used in professional laboratories.
CV% 3.25 1.65 Please consult local regulations for a correct waste disposal.
S56: dispose of this material and its container at hazardous or
special waste collection point.
Methods Comparison S57: use appropriate container to avoid environmental contamination.
S61: avoid release in environment. refer to special instructions/safety
A comparison between Spectrum Diagnostics Urea/BUN reagent data sheets.
and a commercial reagent of the same methodology was performed
on 20 human sera. A correlation of 0.97 was obtained.
References
Sensitivity 1. Batton, C. J & crouch, S.R : Anal. Chem., 1977,49:464-469.
2. Shephard MD, Mezzachi RD : Clin Biochem Revs, 4:61-7, 1983.
When run as recommended, the minimum detection limit of the 3. Tietz NW, ED. Clinical guide to Laboratory tests. 2ND ED.
assay is 0.6 mg/dL. Philadelphia: WB Saunders; 1990:566.
4. Tiffany to, jansen JM, Burtis CA,Overton JB, Scott CD. Enzymatic
Linearity Kinetic Rate and end Point analyses of Substrate, By USE of A
Gemsaec fast analyzer. Clin Chem.
The reaction is linear up to a urea concentration of (200 mg/dl)
33.3 mmol/L. Specimens showing higher concentrations should
be diluted 1+2 with physiological saline and repeat the assay
(result×3). ORDERING INFORMATION
Interfering Substances CATALOG NO. QUANTITY
Haemolysis 318 001 100 Test
Erythrocyte contamination doesn’t elevate results. 318 002 200 Test
318 003 500 Test
Icterus 318 004 1000 Test
No significant interference.
Lipemia
Lipemic specimens interfere with the method of Berthlot.
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
IFUFCC40 Rev.(5), 17/12/2018