Solution reagent BIT series

for clinical chemistrytotal bilirubin

This reagent is intended for the in vitro quantitative determination of Total Bilirubin in human serum. METHODOLOGY This total bilirubin method is based on a modification of the Pearlmen and Lee method in which a surfactant is used as a solubiliser. Sodium nitrite is added to sulfanilic acid to form diazotized sulfanilic acid. Bilirubin in the sample reacts with the diazotized sulfanilic acid to produce azobilirubin which absorbs strongly at 550 nm. The absorbance measured at 550 nm is directly proportional to the total bilirubin concentration in the sample. REAGENT PREPARATION All the reagents are ready to use. REAGENT DETERIORATION Do not use the reagent if: 1. Presence of particles and turbidity. 2. Colour development in R2. NTERFERENCE A comprehensive list of drugs and substances which may interfere with the Total Bilirubin assay. Hemoglobin: No interference up to 500 mg/dL. Lipemia: No interference in the presence of triglycerides up to 1500 mg/dL. Ascorbic Acid is interference. ADDITIONAL EQUIPMENT REQUIRED BUT NOT PROVIDED 1. Spectrophotometer or colorimeter measuring absorbance at 555 ( 530-580)nm. 2. Deionized water and related equipment, e.g.: pipettes 3. Analyzer specific consumables, e.g.: sample cups 4. Control and calibrator materials. Procedure Notes For bilirubin determination in newborns,pipette 10 µL of sample. Multiply the result by 2. CALIBRATION bt products serum calibrator or Bilirubin calibrator can be used . QUALITY CONTROL The integrity of the reaction should be monitored by use of a two level control with known Total Bilirubin values.