Principle
Bilirubin reacts with diazotized sulfanilic acid to produce azobilirubin (violet color). DMSO catalyzes the formation of azobilirubin from free bilirubin, The violet color is proportional to bilirubin concentration measured at 546 nm.(530 – 550 nm.)
Procedure
Reaction type…………………. End-Point
Reaction time………………….5 min at R.T (25-30°C)
Wavelength………………….546 nm. (530-550 nm)
Zero setting with………………….Serum Blank
Sample volume………………….0.05 ml (50 µl)
Reagent volume………………….1.02 ml
Factor………………….20.2
Linearity………………….20 mg%
Quality Control
It’s Recommended that each batch should include a normal and an abnormal commercial reference control serum or a known patient serum, Use of quality control serum, checks both instrument and reagent functions together. Factors which might affect the performance of this test include proper instrument function, temperature control, cleanliness of glassware. Accuracy of pipetting and serum exposure to light.
Other Details
The reagent kit should be stored at Room Temperature (25-30°C) and is stable till the expiry date indicated on the label.
The working solution is stable for 2 days at 2- 80 and for 5 hours at Room Temperature (25-30°C).
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Delicia Adolfson –
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