BIOLABO

www.biolabo.fr
UREA Colorimetric Method
MANUFACTURER:
BIOLABO SAS,
Reagent for quantitative determination of urea
Les Hautes Rives in human serum and plasma or urines.
02160, Maizy, France

REF 80221 R1 1 x 125 mL R2 1 x 1.25 mL R3 1 x 31 mL R4 1 x 10 mL

REF 80321 R1 1 x 500 mL R2 1 x 5 mL R3 1 x 125 mL R4 1 x 10 mL

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TECHNICAL SUPPORT AND ORDERS
Tel : (33) 03 23 25 15 50
IVD IN VITRO DIAGNOSTIC USE
Fax: (33) 03 23 256 256

CLINICAL SIGNIFICANCE (1) (5)

REAGENTS PREPARATION
More than 90% of urea is excreted through the kidneys in urines. Add contents of vial R2 (Urease) into vial R1 (Salicylate). Mix gently by
Measurement of the plasma or serum urea concentration is widely inversion.
regarded as a test of renal function. However, a number of nonrenal
factors also influence the circulating urea concentration: Urea Alkaline (vial R3):
increased level occurs when proteins catabolism is accelerated, burns, Procedure n°1 and n°2 (manual): Dilute (1 + 3) with demineralised
stress, myocardial infarction... Urea is decreased in acute liver
water
destruction and is accompanied with increased ammonium level. Urea
level is generally studied in conjunction with creatinine level Procedure n°3 (manual or automatic): Ready to use
(urea/creatinine ratio) to refine post-renal or pre-renal diagnosis.
Standard (vial R4): transfer the requested quantity, recap and store at
2-8°C.
PRINCIPLE (4)
Enzymatic and colorimetric method based on the specific action of
urease which hydrolyses urea in ammonium ions and carbon dioxide. STABILITY AND STORAGE
Ammonium ions then form with chloride and salicylate a blue-green Store at 2-8°C, well recap in the original vial and away from light.
complex. This coloration, proportional to urea concentration in the • Unopened, reagents are stable until expiry date stated on the label
specimen, is measured at 600 nm. when stored and used as described in the insert.
• Once reconstituted, working reagent (R1+R2) is stable for 1 month
REAGENTS COMPOSITION when free from contamination.
• Once opened, Base (vial R3) diluted ¼ is stable for 3 months when
Vial R1 SALICYLATE free from contamination.
• Once opened, the contents of vial R4 is stable for at least 3 months
Salicylate 31 mmol/L
when free from contamination.
Nitroprussiate 1.67 mmol/L
Discard any reagent if cloudy or if absorbance of blank against water
Vial R2 UREASE at 600 nm > 0.100.
Urease > 15 KUI/L Don’t use working reagent or diluted contents of vial R3 after expiry
date stated on the label of the kit.
Vial R3 BASE
SPECIMEN COLLECTION AND HANDLING (2)
Sodium hypochlorite 7 mmol/L
Sodium hydroxide 62 mmol/L Unhemolysed serum or heparinised plasma. Avoid fluoride or
Before dilution: Xi, R36/38, Irritating to eyes and skin
ammonium as anticoagulant which interfere with the assay.
S24-25-26-28: Avoid contact with skin and eyes. After contact with skin, rinse Urea is stable in serum or plasma for:
immediatly with plenty of water. After contact with eyes, rinse immediatly with plenty of • 24 h at room temperature.
water and seek medical advice
• several days at 2-8°C.
Once diluted: None
• at least 2-3 months frozen.
Vial R4 STANDARD Urea 40 mg/dL (6.66 mmol/L) 24h Urine: diluted (1+19) with demineralised water before assay.
Urea is stable in urines for:
• 4 days at 2-8°C.
SAFETY CAUTIONS Add antibacterial agent as Thymol to improve the stability.
BIOLABO reagents are designated for professional, in vitro diagnostic
use.
INTERFERENCES (3)
• Verify the integrity of the contents before use.
• Use adequate protections (overall, gloves, glasses). No interference of assayed substances (ascorbic acid, bilirubin,
haemoglobin, triglycerides) with the test.
• Do not pipette by mouth.
• In case of contact with skin or eyes, thoroughly wash affected areas For a more comprehensive review of factors affecting this assay refer
with plenty of water and seek medical advice. to the publication of Young D.S.
• Reagents contain sodium azide (concentration < 0.1%) which may
react with copper and lead plumbing. Flush with plenty of water
when disposing. MATERIAL REQUIRED BUT NOT PROVIDED
• Material Safety Data Sheet is available upon request. 1. Basic medical analysis laboratory equipment.
• Waste disposal: Respect legislation in force in the country. 2. Normal and pathological control sera
All specimens should be handled as potentially infectious, in
accordance with good laboratory practices using appropriate
precautions. Respect legislation in force in the country.

IVD REF LOT →

Manufacturer Use by In vitro diagnostic Temperature limitation Catalogue number See insert Batch number Store away from light sufficient for dilute with

Made in France Latest revision : www.biolabo.fr Revision : 26/07/2011

CALIBRATION LINEARITY
• Standard enclosed in the kit (vial R4) or BIOLABO Multicalibrator Procedure n°1 and n°3:
REF 95015 traceable to SRM 909b. The reaction is linear up to 250 mg/dL (41.7 mmol/L).
• Or any calibrator traceable to a reference method or material. Procedure n°2: The reaction is linear up to 125 mg/dL (20.9 mmol/L).
Above, dilute the specimen with saline solution and reassay taking into
The calibration frequency depends on proper instrument functions and
account dilution factor to calculate the result. Linearity limit depends on
on the preservation of reagent.
specimen/reagent ratio.
It is recommended to calibrate in the following cases :
1. When changing batch of reagent. MANUAL PROCEDURE
2. After maintenance operations on the instrument.
3. If control values are out of range, even after using a new vial of Let stand reagents and specimens at room temperature.
fresh control serum. Procedure n°1

Pipette into test tubes Blank Standard Assay

QUALITY CONTROL
Working reagent (R1+R2) 1 mL 1 mL 1 mL
• BIOLABO EXATROL-N Level I REF 95010.
Demineralised water
• BIOLABO EXATROL-P Level II REF 95011. 5 µL

• Other assayed control sera referring to the same method. Standard 5 µL

• External quality control program. Specimen (Note 1) 5 µL

It is recommended to control in the following cases: Mix and wait for 4 minutes at room temperature or 2 minutes at 37°C
• At least once a run. 1 mL 1 mL 1 mL
Base (vial R3) diluted ¼
• At least once within 24 hours.
• When changing vial of reagent. Mix. Let stands for 8 minutes at room temperature or 5 minutes at 37°C. Read absorbance at
• After maintenance operations on the instrument. 600 nm (590-610) against blank (Note 3). Reaction coloration is stable for 2 hours.
If control is out of range, apply following actions:
Procedure n°2
1. Repeat the test with the same control.
2. If control is still out of range, prepare a fresh control and repeat Pipette into test tubes Blank Standard Assay
the test. Working reagent (R1+R2) 1 mL 1 mL 1 mL
3. If control is still out of range, use a new vial of calibrator or a fresh
Demineralised water 10 µL
calibrator and repeat the test.
4. If control is still out of range, calibrate with a new vial of reagent. Standard 10 µL
5. If control is still out of range, please contact BIOLABO technical Specimen (Note 1) 10 µL
support or your local Agent.
Mix and wait for 4 minutes at room temperature or 2 minutes at 37°C

Base (vial R3) diluted ¼ 1 mL 1 mL 1 mL

EXPECTED VALUES (2)
In serum and plasma mg/dL [mmol/L] Mix. Let stands for 8 minutes at room temperature or 5 minutes at 37°C. Read absorbance at
600 nm (590-610) against blank (Note 3). Reaction coloration is stable for 2 hours.
In cord 45-86 [7.5-14.3]
Premature 6-54 [1.1-8.9] Procédure n°3 (alcalin PUR)

< 1 year 9-41 [1.4-6.8] Pipette into test tubes Blanc Etalon Dosage
Working reagent (R1+R2) 1 mL 1 mL 1 mL
Children 11-39 [1.8-6.4]
Demineralised water 5 µL
18-60 years 13-43 [2.1-7.1] Standard 5 µL

60-90 years 17-49 [2.9-8.2] Specimen (Note 1) 5 µL

> 90 years 21-66 [3.6-11.1] Mix and wait for 4 minutes at room temperature or 2 minutes at 37°C

Base (vial R3) PUR 250 µL 250 µL 250 µL

In urines 26-43 g/24 h [0.43-0.71 mol/24 h]
Mix. Let stands for 8 minutes at room temperature or 5 minutes at 37°C. Read absorbance at
To calculate blood urea nitrogen (BUN): multiply the value of urea 600 nm (590-610) against blank (Note 3). Reaction coloration is stable for 2 hours.
(mg/dL) by 0.467.
Each laboratory should establish its own normal ranges for the 1. Serum, plasma or urines diluted (1+19) with demineralised water.
population that it serves. 2. Specific procedures are available upon request for automated
instruments. Please contact BIOLABO technical support.
3. The test may be performed at 578 nm. In this case, Procedure n°2 is
PERFORMANCE CHARACTERISTICS (Procedure n°1) linear up to 300 mg/dL.

Within run Normal level High level Between run Normal level High level CALCULATION
N = 20 N = 20
Calculate the result as follows:
Mean mg/dL 40 141.6 Mean mg/dL 35 111 Serum and plasma:
S.D. mg/dL 0.76 1.66 S.D. mg/dL 1.58 3.44 Abs (Assay)
Result = x Standard concentration
C.V. % 1.89 1.17 C.V. % 4.5 3.1 Abs (Standard)
Detection limit: approximately 10 mg/dL. Urines diluted (1+19):
Multiply the result by 20 (dilution factor).
Comparison with commercially available reagent: y = 0.9816 + 0.87
r = 0.9961 REFERENCES
rd
Sensitivity for 100 mg/dL at 600 nm (1) TIETZ N.W. Text book of clinical chemistry, 3 Ed. C.A. Burtis, E.R.
Ashwood, W.B. Saunders (1999) p. 1239-1241.
Procedure n°1 Approx. 0.400 abs (2) Clinical Guide to Laboratory Test, 4th Ed., N.W. TIETZ (2006) p. 1096-
Procedure n°2 Approx. 0.800 abs 1099.
(3) YOUNG D.S., Effect of Drugs on Clinical laboratory Tests, 4th Ed. (1990)
Procedure n°3 Approx. 0.700 abs p. 3-599 to 3-609
(4) SEARCY R.L., REARDON J.E., FOREMAN J.A., Amer. J. Méd. Techn.
Note: Sensitivity is higher at upper wavelength and lower at inferior 1967, 33, 15-20.
ème
wavelength. (5) Bernard S. Bioch. clin. Diagnostics médicaux chirurgicaux 2 éd. p.143-
144. Ed. Maloine PARIS (1989).
(4) SRM: Standard Reference Material ®

Made in France Latest revision : www.biolabo.fr Revision : 26/07/2011