• Method –Jendrassik & Grof
• Ready-to-use liquid stable reagents
• Simple test procedure : 1.0 ml + 0.02 ml + 0.05 ml
• Results via factor : 20.2 at 530 nm. – 550 nm.
• Autozyme Bilirubin is a reagent set for determination of Total and Direct Bilirubin based on Jendrassik & Grof method, using Diazotized Sulfanilic Acid with DMSO as an activator for
• Total Bilirubin
• Autozyme Bilirubin is a two reagent system using one procedure
• Autozyme Bilirubin is linear up to 20mg%
• Autozyme Bilirubin has one step reconstitution
• Autozyme Bilirubin can be determined in just 5 minutes
• Autozyme Bilirubin method is relatively free of interference from hemoglobin and other commonly occurring substances in serum or plasma

Principle :

Bilirubin reacts with diazotized sulfanilic acid in acidic medium to
produce azobilirubin. DMSO catalyzes the formation of azobilirubin
from free bilirubin. DMSO
Bilirubin + Sulphanilic acid + Sodium NitriteAzobilirubin
Direct bilirubin, being water soluble directly reacts in acid c med um to produce Azobilirubin.
Bilirubin Sulphanilic acid + Sodium Nitrite Azobilirubin
The intensity of the viciet co our formed is directly proportional to the Concentration of bilirubin in the specimen and is measured
photometrically.

Procedure  :

Reaction type……………………. End-Point E
Reaction time ………………….5 mins. at R.T. (25-30°C)
Wavelength……………………………546 nm.(530 – 550 nm.)
Zero setting with…………………….Serum Blank
Sample volume ……………………… 0.05 ml (50 III)
Reagent volume…………………..1.02 ml
Factor …………………………………20.2
Linearity …………………………20 mg%

Quality Control :

Each batch of AutoPure Bilirubin is assayed with at least six quality control sera – Precinom Precipath*, Bio Rad |” Biorad It* Accutrol Normal and Accutrol Abnormal prior to release
To ensure adequate quality control, it is recommended that the laboratory should Use a normal and abnormal commercial reference control serum. It should Be realized that the use of quality control material checks both the reagent and
instrument functions together.
H the control values fall outside the specified limits, each of the below criteria should be cross checked and corrected:

-Proper instrument function wavelength setting, light sourCe and temperature
-Cleanliness of probes anc cuvettes.
-Bacterial contamination of wash water used by the instrument
-Expiry date of the reagent kt.
-Brand naTIE of Roche Diagnostics, CermaryBrand name a Biorad. USA,subiect to
availability of control serum.