Detection of bilirubin conjugates in faeces of germfree rats

The presence of bilirubin conjugates in faeces from germfree (GF) and conventional (CONV) rats was tested after using affinity chromatography and Porapak Q chromatography as clean-up procedures. The bilirubin conjugates were detected as their ethyl anthranilate azopigments after separation by thin layer chromatography (tlc). Azopigments prepared from bile of GF and CONV rats served as a reference material for the tlc analysis. After tlc of the faecal azopigment preparations it was concluded that GF rat faeces contains bilirubin conjugates, while faeces from CONV rats is devoid of conjugated bilirubin. The findings suggest that the main bilirubin conjugates present in faeces from GF rats are of the glucuronic acid type.