UREA

LIQUID STABLE REAGENT IVD

UREASE - GLDH - FIXED TIME

For in vitro Diagnostic use only

INTENDED USE WORKING REAGENT STABILITY
This reagent is intended for in vitro determination of Urea in The working reagent (4R1 : 1R2) is stable for 30 days at 2...8°C,
serum / plasma. when protected from contamination and light. It is recommended to
CLINICAL SIGNIFICANCE prepare fresh working solution before assay is performed.
Urea is the major end product of protein metabolism in humans.
It constitutes the largest fraction of the non-protein nitrogen com- INDICATIONS OF REAGENT DETERIORATION
ponent of the blood. Urea is produced in the liver and excreted - Turbidity
through the kidneys in the urine. Consequently the circulating - Working Reagent absorbance <0.8 at 340 nm.
levels of urea depend upon protein intake, protein catabolism - Failure to recover control values within the acceptable range.
and kidney function.
INCREASES SAMPLE
Elevated serum urea concentrations are observed in impaired Unhaemolysed serum, EDTA or heparinised plasma. Do not use
kidney function, liver diseases, congestive cardiac failure, diabe- heparin ammonium salt and fluoride as anticoagulants. Urea in
tes, infections and diseases which impair kidney function. serum is stable for 7 days at 2...8°C. It is recommended to per-
form assay with freshly collected samples.
METHODOLOGY
Kinetic, enzymatic method. Talke and Schubert, Tiffany et al.
ASSAY PARAMETERS
PRINCIPLE
Urease Mode Fixed time
Urea + 2H2O 2 NH4+ + CO32- Wavelength 1 (nm) 340
GLDH Wavelength 2 (nm) --
NH4+ + 2-Oxoglutarate + NADH L-Glutamate + NAD+ + H2O
Sample Volume (µl) 5 / 10
The rate of absorbance changing at 340 nm is directly proportional Reagent Volume (µl) 500 / 1000
to Urea concentration in the serum.
Lag time (sec.) 20
REAGENT COMPOSITION
(Concentration and Activity in the test) Kinetic interval (sec.) 60
Active Ingredient R1 Concentration No. of readings 1
Tris Buffer (pH 8) 100 mmol/l Reaction temperature (°C) 37
Urease ≥ 10 kU/l Reaction direction Decreasing
GLDH ≥ 2.5 kU/l Normal low (mg/dl) 13
2-Oxoglutarate 5.49 mmol/l Normal high (mg/dl) 45
Active Ingredient R2 Concentration Linearity low (mg/dl) 0
NADH 1.66 mmol/l Linearity high (mg/dl) 250
Also contains Non-reactive fillers and stabilizers. Abs. limit (Max) 1.1
UREA Calibrator Concentration of Standard (mg/dl) See bottle label
Urea Calibrator See bottle label Blank with Reagent
Units mg/dl
WARNING
For in vitro diagnostic use. To be handled by entitled and professi- Programme parameters for specific clinical analysers are
onally educated person. available on request.
Reagents of the kit are not classified like dangerous but contain
less than 0.1% sodium azide - classified as very toxic and dange- ASSAY PROCEDURE
rous substance for the environment.
Pipette into tubes marked Calibrator Test

WORKING REAGENT PREPARATION Working Reagent 1000 µl 1000 µl

The reagent (R1 & R2) are ready to use. The working reagent is Calibrator 10 µl --
prepared by mixing gently 4 parts of R1 with 1 part of R2.
Test -- 10 µl
STORAGE AND STABILITY Mix well and aspirate calibrator followed by samples. Read initial
Unopened reagent bottles (R1 and R2) and Standard are stable till absorbance (A1) 20 seconds after mixing and final absorbance ( A2)
the expiry date stated on the label when stored at 2...8°C. 80 seconds after mixing.

QUALITY SYSTEM CERTIFIED

ISO 9001 ISO 13485
CALCULATION REFERENCES
Calculate the results as follows: 1. Kassirer J. P. : New Eng. J. Med. 285, 385 (1971).
ΔA = A2 - A1 2. Talke H. N., Schubert G. E. : Klin. Wschr. 42, 174 (1965).
Δ A of Test 3. Mackay E. M., Mackay L. L. : Clin. Invest. 4, 295 (1927).
Urea (mg/dl) = x Concentration of Calibrator
4. Sarre H. : Nierenkrankheiten. George Thieme Verlag, Stuttgart
Δ A of Standard
(1959).
UNIT CONVERSION 5. Tietz NW, ed. Clinical Guide to Laboratory Tests, 3rd ed. Phila-
mg/dl x 0.166 = mmol/l delphia, PA : WB Saunders, (1995).
6. Young D. S., Effects of Preanalytical Variables on Clinical Labo-
REFERENCE VALUES 5 ratory Tests, 1st ed. Washington, DC : AACC Press, 3-306 (1995).
Serum / Plasma 13 - 45 mg/dl 7. Burtis C.A., Ashwood E R, ed. Tietz Textbook of Clinical Che-
mistry, 2nd ed. Philadelphia PA : WB Saunders, 2209 (1994).
It is recommended for each laboratory to establish its own reference
8. Dembinska - Kiec A, Naskalski IW : Diagnostyka Laboratoryjna
range for the population it serves.
z elementami biochemii klinicznej, Volumed, 24 - 25 (1998).
9. Kaplan, L. A., Pesce., A. J. : Clinical Chemistry. Theory, ana-
PERFORMANCE CHARACTERISTICS
These characteristics have been obtained using an automatic lysis and correlation 3rd ed., the C. V. Mosby Company, St. Louis
analyzer. Results may vary if a different instrument or a manual 1996, p. 499.
procedure is used. PACK PRESENTATION
Product Pack UREA UREA UREA
LINEARITY Code Size Reagent 1 Reagent 2 Calibrator
Up to 250 mg/dl. For higher values it is recommended to dilute
the samples with normal saline and repeat the assay. Multiply the FBCEM0012 5 x 30 ml 4 x 30 ml 1 x 30 ml 1 x 5 ml
result with the dilution factor.
FBCEM0013 5 x 60 ml 4 x 60 ml 1 x 60 ml 1 x 5 ml

SENSITIVITY
Version No.: EL - 9 - FBCEM - UREA
Limit of Quantitation – 7.0 mg/dl
Date of Issue: 1. 6. 2011

SPECIFICITY / INTERFERENCE SYMBOLS

Haemoglobin up to 5 g/l, Bilirubin up to 30 mg/dl and Triglycerides The following symbols are used in the labeling of ERBA Mannheim
up to 10 g/l do not interfere with the test. kits:

PRECISION
Repeatibility (within run) n Mean (mg/dl) SD (mg/dl) CV (%)
Level 1 20 42 1.4 3.3
Level 2 20 137 2.6 1.9

Reproducibility (between run) n Mean (mg/dl) SD (mg/dl) CV (%)

Level 1 25 42 1.8 4.3
Level 2 25 137 3.8 2.8
UREA
METHOD COMPARISON
A comparison between ERBA reagent (y) and commercially
available assay was performed. The comparison did not show
systematic differences. Details of the comparison experiments
are available on request.

WASTE MANAGEMENT
As per local legal requirements.