Total billirubin couples with diazotised sulphanillic acid in the presence of TAB to form pink coloured azobillirubin complex of both Direct & Indirect billirubin & in the absence of TAB only direct billirubin reacts with diazotised sulphanillic acid to form azobillirubin complex. The intensity of the colour formed is directly proportional to the billirubin present in the sample.
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