Urea FS*

Diagnostic reagent for quantitative in vitro determination of urea in serum, plasma or urine on photometric
systems

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Cat. No. Kit size Please refer to local legal requirements.
1 3101 99 10 021 R1 4x 20 mL + R2 1x 20 mL Reagent Preparation
+ 3 mL Standard
1 3101 99 10 026 R1 5x 80 mL + R2 1x 100 mL Substrate Start
1 3101 99 10 023 R1 1x 800 mL + R2 1x 200 mL
The standard and the reagents are ready to use.
1 3101 99 10 704 R1 8x 50 mL + R2 8x 12.5 mL
1 3101 99 10 917 R1 8x 60 mL + R2 8x 15 mL Sample Start
1 3101 99 90 314 R1 10 x 20 mL + R2 2x 30 mL Mix 4 parts of R1 with 1 part of R2
1 3100 99 10 030 6x 3 mL Standard
(e.g. 20 mL R1 + 5 mL R2) = mono reagent
Summary [1,2] Leave the mono reagent for at least 30 min. at 15 – 25°C before
use.
Urea is the nitrogen-containing end product of protein catabolism. Stability: 4 weeks at 2 – 8°C
States associated with elevated levels of urea in blood are referred 5 days at 15 – 25°C
to as hyperuremia or azotemia. Parallel determination of urea and Protect mono reagent from light!
creatinine is performed to differentiate between pre-renal and post-
renal azotemia. Pre-renal azotemia, caused by e.g. dehydration, Materials required but not provided
increased protein catabolism, cortisol treatment or decreased renal NaCl solution 9 g/L
perfusion, leads to increased urea levels, while creatinine values General laboratory equipment
remain within the reference range. In post-renal azotemias, caused
by the obstruction of the urinary tract, both urea and creatinine Specimen
levels rise, but creatinine in a smaller extent. In renal diseases Serum, plasma (no ammonium heparin!), fresh urine
urea concentrations are elevated when the glomerular filtration Dilute urine 1 + 50 with dist. water and multiply results by 51.
rate is markedly reduced and when the protein intake is higher TruLab Urine controls must be prediluted the same way as patient
than 200 g/day. samples.
Method Stability [4]
in serum or plasma:
“Urease – GLDH“: enzymatic UV test
7 days at 20 – 25°C
Principle 7 days at 4 – 8°C
1 year at –20°C
Urea + 2 H2O Urease > 2 NH4+ + 2 HCO3-
in urine:
2 days at 20 – 25°C
2-Oxoglutarate + NH4+ + NADH GLDH > L-Glutamate + NAD+ + H2O
7 days at 4 – 8°C
1 month at –20°C
GLDH: Glutamate dehydrogenase
Freeze only once! Discard contaminated specimens.
Reagents
Assay Procedure
Components and Concentrations
R1: TRIS pH 7.8 150 mmol/L Application sheets for automated systems are available on
2-Oxoglutarate 9 mmol/L request.
ADP 0.75 mmol/L Wavelength 340 nm, Hg 334 nm, Hg 365 nm
Urease ≥ 7 kU/L Optical path 1 cm
GLDH (Glutamate dehydrogenase, bovine) ≥ 1 kU/L Temperature 25°C/30°C/37°C
R2: NADH 1.3 mmol/L Measurement Against reagent blank
Standard: 50 mg/dL (8.33 mmol/L) 2-point kinetic

Storage Instructions and Reagent Stability Substrate start

Reagents and standard are stable up to the end of the indicated Blank Sample or standard
month of expiry, if stored at 2 – 8°C, protected from light and Sample or standard - 10 µL
contamination is avoided. Do not freeze the reagents! Reagent 1 1000 µL 1000 µL
Mix, incubate 0 – 5 min., then add:
Warnings and Precautions Reagent 2 250 µL 250 µL
1. The reagents contain sodium azide (0.95 g/L) as Mix, incubate for approx. 60 sec. at 25°C/30°C or approx.
preservative. Do not swallow! Avoid contact with skin and 30 – 40 sec at 37°C, then read absorbance A1. Read absorbance
mucous membranes. A2 exactly after another 60 seconds.
2. Reagent 1 contains animal material. Handle the product as ∆A = (A1 – A2) sample or standard
potentially infectious according to universal precautions and
good laboratory practice. Sample start
3. In very rare cases, samples of patients with gammopathy Blank Sample or standard
might give falsified results [6]. Sample or standard - 10 µL
4. Please refer to the safety data sheets and take the necessary Mono reagent 1000 µL 1000 µL
precautions for the use of laboratory reagents. For diagnostic Mix, incubate for approx. 60 sec. at 25°C/30°C or approx.
purposes, the results should always be assessed with the 30 – 40 sec at 37°C, then read absorbance A1. Read absorbance
patient’s medical history, clinical examinations and other A2 exactly after another 60 seconds.
findings.
∆A = (A1 – A2) sample or standard
5. For professional use only!

Urea FS – Page 1 * fluid stable

Notes Inter-assay precision Mean SD CV
1. The method is optimized for 2-point kinetic measurement. It is n = 20 [mg/dL] [mg/dL] [%]
recommended to perform the method only on mechanized Sample 1 20.3 0.58 2.88
equipment because it is difficult to incubate all samples and the Sample 2 48.3 1.12 2.32
reagent blank strictly for the same time intervals. The assay Sample 3 152 1.38 0.91
scheme may be used for adaptation purposes for instruments
Method Comparison
with no specific adaptation sheet. The volumes may be
proportionally smaller. A comparison of DiaSys Urea FS (y) with a commercially available
2. The statement “approx. 60 sec. or approx. 30 - 40 sec“ means test (x) using 68 samples gave following results:
that the time period chosen does not need to be exactly 60 resp. y = 0.99 x + 1.06 mg/dL; r = 0.999
30 – 40 sec. A time period once chosen (e.g. 55 sec.) has to be
respected exactly for all samples, standards and the reagent Reference Range
blanc. In Serum/Plasma [1]
Calculation [mg/dL] [mmol/L]
Adults
With standard or calibrator Global 17 – 43 2.8 – 7.2
∆A Sample Women < 50 years 15 – 40 2.6 – 6.7
Urea [mg/ dL] = × Conc. Std/ Cal [mg / dL] Women > 50 years 21 – 43 3.5 – 7.2
∆A Std/ Cal
Men < 50 years 19 – 44 3.2 – 7.3
Conversion factor Men > 50 years 18 – 55 3.0 – 9.2
Urea [mg/dL] x 0.1665 = Urea [mmol/L] Children
1 – 3 year(s) 11 – 36 1.8 – 6.0
Urea [mg/dL] x 0.467 = BUN [mg/dL] 4 – 13 years 15 – 36 2.5 – 6.0
14 – 19 years 18 – 45 2.9 – 7.5
BUN [mg/dL] x 2.14 = Urea [mg/dL]

(BUN: Blood urea nitrogen) BUN in Serum/plasma [mg/dL] [mmol/L]

Adults
Calibrators and Controls Global 7.94 – 20.1 2.8 – 7.2
For the calibration of automated photometric systems, DiaSys Women < 50 years 7.01 – 18.7 2.6 – 6.7
TruCal U calibrator is recommended. The assigned values of the Women > 50 years 9.81 – 20.1 3.5 – 7.2
calibrators have been made traceable to NIST SRM®-909 Level 1. Men < 50 years 8.87 – 20.5 3.2 – 7.3
DiaSys TruLab N, P and TruLab Urine controls should be assayed Men > 50 years 8.41 – 25.7 3.0 – 9.2
for internal quality control. Each laboratory should establish
Children
corrective action in case of deviations in control recovery.
Cat. No. Kit size 1 – 3 year(s) 5.14 – 16.8 1.8 – 6.0
TruCal U 5 9100 99 10 063 20 x 3 mL 4 – 13 years 7.01 – 16.8 2.5 – 6.0
5 9100 99 10 064 6 x 3 mL 14 – 19 years 8.41 – 21.0 2.9 – 7.5
TruLab N 5 9000 99 10 062 20 x 5 mL Urea/Creatinine ratio in serum [1]
5 9000 99 10 061 6 x 5 mL 25 – 40 [(mmol/L)/(mmol/L)]
TruLab P 5 9050 99 10 062 20 x 5 mL 20 – 35 [(mg/dL)/(mg/dL)]
5 9050 99 10 061 6 x 5 mL
TruLab Urine Level 1 5 9170 99 10 062 20 x 5 mL Urea in Urine [2]
5 9170 99 10 061 6 x 5 mL 26 – 43 g/24h (0.43 – 0.72 mol/24h)
TruLab Urine Level 2 5 9180 99 10 062 20 x 5 mL
5 9180 99 10 061 6 x 5 mL Each laboratory should check if the reference ranges are
transferable to its own patient population and determine own
Performance Characteristics reference ranges if necessary.
Measuring range Literature
The test has been developed to determine urea concentrations 1. Thomas L. Clinical Laboratory Diagnostics. 1st ed. Frankfurt:
within a measuring range from 2 – 300 mg/dL (0.3 – 50 mmol/L) in TH-Books Verlagsgesellschaft; 1998. p. 374-7.
serum/plasma respectively up to 30 g/dL (5 mol/L) in urine. When 2. Burtis CA, Ashwood ER, editors. Tietz Textbook of Clinical
values exceed this range the samples should be diluted 1 + 2 with Chemistry. 3rd ed. Philadelphia: W.B Saunders Company;
NaCl solution (9 g/L) and the result multiplied by 3. 1999. p. 1838.
Specificity/Interferences 3. Talke H, Schubert GE. Enzymatische Harnstoffbestimmung in
Blut und Serum im optischen Test nach Warburg (Enzymatic
No interference was observed by ascorbic acid up to 30 mg/dL,
determination of urea in blood and serum with the optical test
bilirubin up to 40 mg/dL, hemoglobin up to 500 mg/dL and lipemia
according to Warburg). Klin Wschr 1965; 43: 174-5.
up to 2000 mg/dL triglycerides. Ammonium ions interfere;
4. Guder WG, Zawta B et al. The Quality of Diagnostic Samples.
therefore, do not use ammonium heparin as anticoagulant for
1st ed. Darmstadt: GIT Verlag; 2001; p. 48-9, 52-3.
collection of plasma! For further information on interfering
5. Young DS. Effects of Drugs on Clinical Laboratory Tests. 5th
substances refer to Young DS [5].
ed. Volume 1 and 2. Washington, DC: The American
Sensitivity/Limit of Detection Association for Clinical Chemistry Press 2000.
The lower limit of detection is 2 mg/dL. 6. Bakker AJ, Mücke M. Gammopathy interference in clinical
chemistry assays: mechanisms, detection and prevention.
Precision (at 37°C) Clin Chem Lab Med 2007; 45(9):1240–1243.
Intra-assay precision Mean SD CV
n = 20 [mg/dL] [.mg/dL] [%] Manufacturer
Sample 1 21.3 0.50 2.33 DiaSys Diagnostic Systems GmbH
Sample 2 35.3 0.82 2.33 IVD Alte Strasse 9 65558 Holzheim Germany
Sample 3 141 1.52 1.08

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