SAFETY PRECAUTIONS AND WARNINGS

For in vitro diagnostic use only. Do not pipette by mouth.

Exercise the normal precautions required for handling laboratory
reagents.

UREA
Enzymatic Kinetic Method
MANUAL
INTENDED USE
For the quantitative in vitro determination of Urea in serum,
plasma and urine. This product is suitable for Manual use.
Cat. No.
UR 10042

R1a. Buffer
R1b. Enzyme Reagent
CAL. Standard

2 x 50 ml
2 x 50 ml
1 x 5.5 ml

UV METHOD (1, 2)
Urea is hydrolyzed in the presence of water and urease to
produce ammonia and carbon dioxide. The ammonia produced
in the first reaction combines with -oxoglutarate and NADH in
the presence of glutamate dehydrogenase to yield glutamate and
NAD+.
PRINCIPLE

Solutions R1a and CAL contains Sodium Azide. Avoid ingestion

or contact with skin or mucous membranes. In case of skin
contact, flush affected area with copious amounts of water. In
case of contact with eyes or if ingested, seek immediate medical
attention.
Sodium Azide reacts with lead and copper plumbing, to form
potentially explosive azides. When disposing of such reagents
flush with large volumes of water to prevent azide build up.
Exposed metal surfaces should be cleaned with 10% sodium
hydroxide.
Health and Safety data sheets are available on request.
The reagents must be used only for the purpose
intended by suitably qualified laboratory personnel,
under appropriate laboratory conditions.

Urease

Urea + H20

2NH3

CO2

STABILITY AND PREPARATION OF REAGENTS

R1a. Buffer
Contents ready for use. Stable up to the expiry date when
stored at +2 to +8C.

2-oxoglutarate + 2NH4++ 2NADH

GLDH

2L-glutamate + 2NAD+ + 2H2O

SAMPLE
Serum, heparinized plasma or EDTA-plasma, urine.
Dilute urine 1 + 20 with distilled water (result x 21).

R1b. Enzyme Reagent

Reconstitute the contents of one vial of Enzyme Reagent R1b
with a portion of Buffer R1a and then transfer the entire
contents to bottle R1a, rinsing bottle R1b several times.
Stable for 4 weeks at +2 to +8C or 2 days at +15 to +25C.

REAGENT COMPOSITION
Contents

Initial Concentrations of the Solutions

R1a. Buffer
Tris-Buffer
Sodium Azide
R1b. Enzyme Reagent
Urease
GLDH
NADH
Adenosine-5-diphosphate
-oxoglutarate
CAL. Standard

150 mmol/l, pH 7.6

< 0.9 % w/v
15 U/ml
1 U/ml
0.28 mmol/l
2.45 mmol/l
11.7 mmol/l
See lot specific insert

CAL. Standard
Contents supplied ready for use. Stable up to the expiry
date when stored at +2 to +8C.
MATERIALS PROVIDED
Buffer
Enzyme Reagent
Standard
MATERIALS REQUIRED BUT NOT PROVIDED
Randox Assayed Multisera Level 2 (Cat. No. HN 4203)
Randox Assayed Multisera Level 3 (Cat. No. HE 4202)
NOTE
All anticoagulants except ammonia-heparinate can be used.

PAGE 1 OF 2

MANUAL UR 10042
PROCEDURE
Wavelength:
Cuvette:
Temperature:
Measurement:

340 nm (Hg 334nm or Hg 365 nm)

1 cm light path
+37C
against reagent blank

Pipette into cuvette:

Reagent Blank

Standard

Sample

10 l
----1000 l

--10 l
--1000 l

----10 l
1000 l

Distilled Water
Standard
Sample
Reagent (R1)

REFERENCES
1. Kassirer, J.P. New Eng. J. Med. 1971; 285: 385.
2. Teitz, N.W., Fundementals of Clinical Chemistry W.B.
Saunders Company, 1970 Philadelphia.
3. Young, D.S., Pestaner, L.C., Gibbermann, V. Clin. Chem.,
1975; 21: 1D.
4. Mackay, E. M., and L. L. Mackay. (1927). J. Clin. Invest. 4: 295.
5. Sarre, H. (1959). Nierenkraukheiten. Georg. Thieme Velag,
Stuttgart.

Mix, read initial absorbance after 30 sec and start timer

simultaneously. Read again after 1 min.
CALCULATION
Asample
Urea Concentration =
(mmol/l)

x
Astandard

Standard
Concentration

QUALITY CONTROL
Randox Assayed Multisera, Level 2 and Level 3 are recommended
for daily quality control.
NORMAL VALUES
Serum (4)
Urine (5)

1.7 - 8.3 mmol/l

333-583 mmol/24 hrs

10 - 50 mg/dl
20 - 35 g/24hrs

It is recommended that each laboratory establish its own

reference range to reflect the age, sex, diet and geographical
location of the population.
LINEARITY
The method is linear to 50.0 mmol/l (300 mg/dl) in serum or
plasma, and 1050 mmol/l (6300 mg/dl) in urine. Samples with
values higher than these concentrations should be diluted 1+1
with 0.9% NaCl solution and the assay repeated multiplying the
results by 2.

14 Jun 16 ml

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