ACCUCARETM
DIRECT BILIRUBIN
monoreagent
ORDER INFORMATION :
Catalog No: Product Description: Pack Size:
TDBIL 100 T+D BILIRUBIN 2 X 50 ML
TDBIL 200 T+D BILIRUBIN 2 X 100 ML

METHOD

Colorimetric end point test

PRINCIPLE
Total bilirubin reacts in acid enviroment with diazotised sulfanilic acid to give a blue azo compound. The complex formed absorbs at 570 nm.
Direct bilirubin (conjugated) reacts in acid environment with diazotised sulfanilic acid. The complex formed absorbs at 570 nm.

Reagent :
Reagent I : Total Bilirubin Reagent
Reagent II : Total Bilirubin Nitrite Reagent
Reagent III : Direct Bilirubin Reagent
Reagent IV : Direct Bilirubin Nitrite Reagent

SAMPLE: Serum or EDTA plasma.

REFERENCE VALUES
Direct Bilirubin 0 - 0.25 mg/dl
Total Bilirubin 0 - 1.2 mg/dl

The reference values are to be considered as indicative only. Every Laboratory should establish is own normal ranges.


QUALITY CONTROL
Accutestrol N - H


NOTES : Haemolysis may cause falsely low values in direct Bilirubin assays.


REAGENT Tietz, N.W., textbook of clinical Chemistry. Gambino, S.R. Michealson, M.Gambino. S.R.et al, jama, W.B.Saunders Co. (1983). 		REAGENT PREPARATION
Reagents are ready to use.

STABILITY : Till Expiry at R.T.

AUTOMATED PARAMETERS
Wave length 570 nm
Cuvette 1 cm light path
Reaction Type End Point
Reaction Temperature 37°C
Measurement Against sample blank
Sample/Reagent Ratio 1 : 20
Incubation 2 minutes
Factor 19.33
Low Normal Refer reference values
High Normal Refer reference values
Linearity Linearity 25 mg/dl

MANUAL PROCEDURE

PIPETTE INTO TEST TUBE
  TSAMPLE TBLANK DSAMPLE DBLANK
TBIL REAGENT 1 1000 µl 1000 µl - -
TBIL REAGENT 2 50 µl - - -
DBIL REAGENT 1 - - 1000 µl 1000 µl
DBIL REAGENT 2 - - 50 µl -
SAMPLE 50 µl 50 µl 50 µl 50 µl

Mix well and Incubate for 5 mins total Bilirubin & 2 mins for direct Bilirubin at 37°C. Measure final absorbance of the sample (Ac) and blank (Ab) against water.

CALCULATION AND LINEARITY

(Ac - Ab) x 19.33 = mg/dl Direct Bilirubin.

(Ac - Ab) x 19.33 = mg/dl Total Bilirubin.

The method is Linear to a Concentration of 25 mg/dl.
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