UREA (UV)

(UV GLDH METHOD)

INTENDED USE: PROCEDURE :
The reagent kit is intended for the "in vitro" quantitative Pipette into clean dry test tube labeled as Standard (S) and
determination of Urea in serum, Plasma and Urine. Test (T) :

CLINICAL SIGNIFICANCE: Adittion sequence S T

Urea is the end product of he protein metabolism. It is synthesized Working reagent 1 ml 1 ml
in the liver from the ammonia produced by the catabolism of amino Standard 10µl -
acids. It is transported by the blood to the kidneys from where it is
excreted. Increased levels are found in renal diseases, urinary Sample - 10 µl
obstructions, shock, congestive heart failure and burns. Decreased
levels are found in liver failure and pregnancy.
Mix well and read the initial absorbance 1A for the Standard and
the Test after exactly 30 seconds. Read another absorbance2A
PRINCIPLE: of the Standard and the Test exactly 60 seconds later. Calculate
Urease hydrolyzes urea to ammonia and CO2. The ammonia
the change in absorbance DA for both the Standard and
formed further combines with α Ketoglutarate and NADH to form Test.
Glutamate and NAD. The rate of oxidation of NADH to NAD is
measured as a decrease in absorbance in a fixed time, which is For Standard D AS = AS2 - AS1
propotional to the urea concentration in the sample. For Test D AT = AT 2 - AT 1

REACTION: CALCULATION :
Urease D AT
+ x 50
Urea + HO
2 + 2H+ 2NH4 + CO2 Urea (mg/dl) =
D AS
GLDH
+ NORMAL VALUE :
2α-ketoglutarate + 2NH4 + 2NADH 2L-Glutamate +
Serum / Plasma : 10-40 mg/dl
2NAD+ + 2HO2
Urine : 26 to 43 g/24 hrs.

CONTENTS: Urea/Creatinine ratio

Reagent 1 : Urea Enzyme Reagent 25 : 40 [(mmol/L)/(mmol/L)]
Reagent 2 : Urea Substrate Reagent 20 : 35 [(mg/dL)/(mg/dL)]
Reagent 3 : Urea Standard 50 mg/dl Each Laboratory should establish it's own normal range
representing its patient population.
MATERIALS REQUIRED BUT NOT PROVIDED:- LINEARITY :
- Clean & Dry Glassware.
- Laboratory Glass Pipettes or Micropipettes & Tips. The procedure is linear upto 300 mg/dl. If the value exceed this
- Bio-Chemistry Analyzer. limit, dilute the serum with normal saline (NaCl 0.9 %) and repeat
the assay. Multiply result by dilution factor.
SAMPLES:
Serum, plasma, urine. QUALITY CONTROL :
Do not use anticoagulants containing fluoride or ammonium ions. For accuracy it is necessary to run known controls with every
Dilute urine 1+49 with distilled water and multiply results by 50. assay.
LIMITATION & PRECAUTIONS :
PREPARATION OF WORKING REAGENT & STABILITY : The reagents should be handled with caution, avoiding
Working reagent: Mix 4 parts of Reagent 1 + 1 part of Reagent 2 . swallowing and conctact with skin, eyes and mucous
Working reagent is stable for 5 days at 15-25 °C and for 4 weeks at
membranes. The use of the laboratory reagents according to
2-8 °C.
Unopened kit is stable til expiry date mentioned on the kit when good laboratory practice is recommended.
stored at 2-8 °C. Do not freeze the reagents. REFERENCES:
(1) Thomas L. Clinical Laboratory Diagnostics. 1st ed.
GENERAL SYSTEM PARAMETERS: Frankfurt : TH-Books Verlagsgesellschaft;1998 p.374-7.
Reaction Type : Fix Time (Decreasing) (2) Burtis CA, Ashwood ER, editors. Tietz Textbook of Clinical
Wave Length : 340 nm. Chemistry. 3 rd ed. Philadelphia: W.B Saunders Company;
Cuvette Temp : 37°C 1999. p. 1838.
(3)Talke H, Schubert GE. Enzymatische Harnstoff-
DelayTime : 30 Sec.
bestimmung in Blut und Serum im optischen Test nach
Interval Time : 60 Sec.
Warburg (Enzymatic determination of urea in blood and
No. of reading :1 serum with the optical test according to Warburg). Klin
Reagent Volume : 1.0 mL Wschr 1965; 43:174-5.
Sample Volume : 10 µL
CODE NO. PACK SIZE Reagent 1 Reagent 2 Reagent 3
Standard : 50 mg/dl S16C 2x20 ml + 1x10 ml 2x20 ml 1x10 ml 3.0 ml
Zero Setting : Deionised water S16A 4x20 ml + 2x10 ml 4x20 ml 2x10 ml 3.0 ml
Light Path : 1 cm.
IVD
UNIONPROJET S.R.L.S.
EC REP Via Aliprandi 41-20851
Lissone (MB)
www.jas-anz.com.au/register

ISO 9001:2015 ISO 13485:2003