Method for the separation of serum bile pigments by thin-layer chromatography on polyamide

Abstract

A method is proposed to seperate bile pigments by thin-layer choromatography.

Bile pigments are extracted from se­rum by means of n-propanol. The ex­tracts are subjected to chromatographic analysis using a Polyamide thin-layer (250 /z thick) as adsorbent and a mixture of alcohol, pyridine and water as a solvent System. Chromatoplates are sprayed with a solution of diazotized p-nitroaniline and the spots are examined with ultraviolet light.

Cole and allied have separated and identified three different pigments from jaundiced sera. In the samples we have examined the chromatographic analysis on Polyamid thin-layer permits the se­paration of more than three pigments, all fractions reacting immediately with diazotized p-nitroaniline. When exami­ned with ultraviolet light of 365 m/x, all spots show a reddish fluorescence.

This new method is of greater sensitivity than that of any used until now as it allows the detection of bilirubin in amounts down to 50 /xgrs.