A spectrophotometric assay of skin bilirubin in adult Gunn rats.
The present study was undertaken to assess the value of a spectrophotometric method for the assay of bilirubin content in Gunn rat skin. The pigment extraction was performed by incubating skin strips in a barbital-acetate buffered methanol-chloroform mixture at pH 7 for 90 min. The procedure yielded an overall recovery average of 93 +/- 9% which was not influenced when the incubation time was varied from 1 to 2.5 h. The bilirubin concentration in skin extracts was calculated from spectrophotometric absorbance readings and according to the molar extinction coefficient of the pigment in chloroform. Bilirubin appeared to remain stable during the analytical process. The mean bilirubin content as measured by this method was 1.88 mumol/100 g (1.1 mg/100 g). It is concluded that the proposed method is well suited for the investigation of bilirubin skin metabolism.
D000818:Animals; D001663:Bilirubin / Q000032:analysis*; D005069:Evaluation Studies as Topic; D005260:Female; D008297:Male; D010860:Pigments, Biological / Q000302:isolation & purification; D051381:Rats; D011911:Rats, Gunn; D012867:Skin / Q000032:analysis*; D013053:Spectrophotometry
D010860:Pigments, Biological; D001663:Bilirubin