• Auto zyme Bilirubin is a reagent set for determination of Total and Direct Bilirubin based on Jurassic & Grof method, using Diazotized Sulphonic Acid with DMSO as an activator for Total Bilirubin.
• Auto Zyme Bilirubin is a two reagent system using one step procedure.
• Auto Zyme Bilirubin is linear upto 20 mg%.
• Auto Zyme Bilirubin can be used on any Colorimeter, Spectrophotometer, Discrete semi-automated and Automated analyzer. Programmed can be designed for any specific analyzer upon request.
• Auto Zyme Bilirubin has one step reconstitution.
• Auto Zyme Bilirubin can be determined in just 5 minutes.
• Auto Zyme Bilirubin method is relatively free of interference from hemoglobin and other commonly occurring substances in serum or plasma.
• This insert covers technical matter for the complete range of Auto Zyme Bilirubin kits available viz.:

Please read the appropriate matter depending on the kit you have purchased.

PRINCIPLE

Bilirubin reacts with diazotized sulphanilic acid to produce azobilirubin (violet colour). DMSO catalyzes the formation of azobilirubin from free bilirubin. The violet colour is proportional to bilirubin concentration measured at 546 nm.(530 - 550 nm.).

TOTAL BILIRUBIN

Bilirubin + Sulphanilic Acid + Sod. Nitrite DMSO x p Azobilirubin

DIRECT BILIRUBIN

Bilirubin + Sulphanilic Acid + Sod. Nitrite .Azobilirubin

DMSO  Dimethyl Sulphoxide

REAGENT STORAGE & STABILITY

• The reagent kit should be stored at Room Temperature (25-30 degree C) and is stable till the expiry date indicated on the label.
• The working solution is stable for 2 days at 2-8 degree C and for 5 hours at Room Temperature (25-30 degree C).

 SPECIMEN COLLECTION & PRESERVATION

Blood should be collected in a clean dry container. Although serum is preferred plasma can also be used as sample.

Following anticoagulant can be used for plasma separation :

• EDTA 2 mg/ml of blood
• HEPARIN 200 IU/ml of blood

Bilirubin is light sensitive. Avoid exposure of serum or plasma to direct light. Bilirubin in serum and plasma is stable for a day at 2-8 degree C or one month at -10 degree C. Samples should be brought to room temperature before use.

PROCEDURE

• Reaction type: End-Point
• Reaction time: 5 mins. at R.T. (25-30 degree 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%