ACCUCARETM
UREA REAGENT
(BERTHELOT METHOD)
ORDER INFORMATION
Catalog No: Product Description: Pack Size:
URE 100 UREA REAGENT 2 x 50 ml
URE 200 UREA REAGENT 2 x 100 ml

METHOD
Colorimetric end point test

PRINCIPLE
The Berthelot reaction has long been used for the measurement of urea and ammonia. The present method is a modified Berthelot Method. The urea colorimetric procedure is a modification of the Berthelot reaction. Urea is converted to ammonium by the use of urease. Ammonium ions then react with a mixture of salicylate, sodium nitroprusside and hypochlorite to yield a blue-green chromophore. The intensity of the color formed is proportional to the urea concentrtion in the sample.

SAMPLE
Serum or plasma,

REFERENCE VALUES
15 - 50 mg/dl
The reference values are to be considered as indicative only. Every Laboratory should establish its own normal range

QUALITY CONTROL :
Accutestrol N - H

REAGENTS
Reagent I : Urea Enzyme reagent
Reagent II : Urea Color developer
Reagent III : Urea Std : 50 mg/dl

REAGENT PREPARATION
Reagent I : Dissolve contents of Reagent I with the Volume of distilled water as specified on the vial
Reagent II : Ready to use

STABILITY:
Reagent I is stable for 45 days on reconstitution when stored at 2-8°C.

Reagent II is stable for 90 days on opening the vial.
The reagent should be clear & pale yellow
AUTOMATED PARAMETERS
Wave length 578 nm
Measurement Against reagent blank
Reaction Type End point
Cuvette 1 cm light path
Reaction Temperature 37°C
Reaction direction Increasing
Sample/Reagent Ratio 1: 200
Incubation 5+5 min
No: of Readings 01
Low Normal at 37°C 15 mg/dl
High Normal at 37°C 50 mg/dl
Linearity at 37°C 200 mg/dl

MANUAL PROCEDURE

  BLANK STD SAMPLE
SAMPLE - - 10µl
STANDARD - 10µl -
REAGENT I 1000µl 1000µl 1000µl

Incubate all tubes for 5 mts at 37°C or 10 mts at rt.

REAGENT II 1000µl 1000µl 1000µl

Incubate all tubes for 5 mts at 37°C or 10 mts at rt.
Zero sectrophotometer with the reagent blank at 578 nm.
Read and record absorbance of samples of all tubes.

CALCULATION AND LINEARITY
Abs of Unk / Abs of Std. X Conc. of Std. = mg/dl Urea

NOTES :
The reagent is linear to 200 mg/dl Urea.

REFERENCES Teitz.N.W; Fundamentals of clinical chemistry, Philadelphia, W.B.Saunders & Co,. Philadelphia, p991 (1976)., Talke H. Schubert GE, Klin Wchers., (1965), 43, 174.
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