title: [Purification of conjugated bilirubin from human urine].
abstract: OBJECTIVE
  To develop a simple, practical method with fine yield and purity for separating conjugated bilirubin(Bc) from a sustainable and facile material in order that it could serve as a calibrator for total and direct bilirubin determination.
  
  
  METHODS
  Isolating Bc from urine of patients with obstructive jaundice by CHCl3: CH3CH2OH(1:1, V/V) followed by CNBr activated Sepharose 4B-HAS affinity chromatography, we analyzed the products by the use of Doumas J-G diazo method and HPLC, respectively.
  
  
  RESULTS
  Determined by J-G diazo method, the crude product extracted by CHCl3: CH3CH2OH(1:1, V/V) yields 90.2 +/- 5.4%(n = 3) of urine Bc, the ratio of Bc to total dry weight being 67.9 +/- 6.4%(n = 3). The final product purified by affinity chromatography with the ratio of separation of 1.5, 1.737 and analyzed by HPLC has the yield of 46.9% and purity of 94.7%(BDG of 76.2%, BMG of 18.5%).
  
  
  CONCLUSIONS
  The above data demonstrate that urine conjugated bilirubin can be successfully purified by CHCl3: CH3CH2OH extraction followed by Sephrose-HSA affinity chromatography with sufficient purity and output.
authors.affiliation: School of Basic Medical Sciences, WCUMS, Chengdu 610041, China.
authors.forename: Y
authors.initials: Y
authors.lastname: Guo
authors.forename: M
authors.initials: M
authors.lastname: Chen
authors.forename: Y
authors.initials: Y
authors.lastname: Tu
authors.forename: H
authors.initials: H
authors.lastname: Luo
authors.forename: S
authors.initials: S
authors.lastname: Li
authors.forename: D
authors.initials: D
authors.lastname: Yan
authors.forename: M
authors.initials: M
authors.lastname: Niu
mesh_terms: D001663:Bilirubin / Q000302:isolation & purification* / Q000652:urine*; D002779:Cholestasis / Q000652:urine*; D002846:Chromatography, Affinity; D006801:Humans
chemical_list: D001663:Bilirubin