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The ever-increasing demand for a sensitive, rapid and reliable method for determination of serum bilirubin level has been inciting the interest of the researchers to develop new methods for both laboratory set up and point of care applications. These efforts embrace measurement of different forms of bilirubin, such as, unconjugated (free and albumin bound) bilirubin, conjugated (direct) bilirubin, and total (both conjugated and unconjugated) bilirubin in the serum that may provide critical information useful for diagnosis of many diseases and metabolic disorders. Herein, an effort has been made to provide a broad overview on the subject starting from the conventional spectroscopy based analytical methods widely practiced in the laboratory setup along with the sophisticated instrument based sensitive methods suitable for determination of different forms of bilirubin to various portable low cost systems applicable in point of care (POC) settings. In all these discussions emphasis is given on the novel methods and techniques bearing potential to measure the bilirubin level in biological samples reliably with less technical complexity and cost. We expect that this review will serve as a ready reference for the researchers and clinical professionals working on the subject and allied fields.Deviation of bilirubin concentration from normal level in serum is associated with many diseases and metabolic disorders.Hence, it is a key compound in blood bearing immense diagnostic importance.Rapid and selective bilirubin determination has also prime importance for managing jaundice in neonates.Herein, advances on developing new methods for bilirubin determination have been discussed.