Indirect determination of bilirubin by linear sweep polarography

Summary A polarographic method for the determination of bilirubin is described: In hydroquinone-sodium phosphate buffer (pH 11.2) medium, bilirubin can be rapidly oxidized to a green product, which causes a sensitive polarographic wave at –0.83 V (vs. SCE); the peak current of the wave is linear with the bilirubin concentration over a range of 3×10^–7 to 5×10^–6 mol/l. It has also been found to be more stable than that produced by bilirubin; in general, it can remain unchanged for about 1 h at room temperature and exposure to air. This overcomes the instability of bilirubin for its direct assay to a certain extent. If a small amount of EDTA is added to the solution, the stability of the wave can be further improved. In addition, the characteristics of the wave have been investigated and it has been confirmed to be an adsorptive catalytic wave. The chemical reaction mechanism is discussed and the green product is confirmed to be biliverdin.