Occurrence of polyhydroxy alcohols in the gall bladder bile of a mud-puppy, Salamander necturus

Bile obtained from the gall bladder of Salamander necturus was investigated for the presence of bile alcohols by a combined technique of Thin Layer Chromatography (TLC), Gas Liquid Chromatography (GLC) and Gas Chromatography-Mass Spectrometry (GC-MS). Dried bile salts were hydrolysed with 2.5N KOH in an autoclave at 121 degrees C. Unsaponifiable matter was extracted with ethyl acetate, three times, washed with distilled water, dried with anhydrous sodium sulphate,evaporated to dryness and analysed for its bile alcohols. Analytical GLC revealed the presence of at least thirteen bile alcohols in the mixture. Two major bile alcohols, i.e. 5alpha-anhydrocyprinol and 5alpha-cyprinol constituted about 73% of the total unsaponifiable matter. Others were identified as 3alpha, 7alpha, 12alpha,trihdroxy-5beta-delta23-homocholene,3alpha,7alpha,12alpha-trihydroxy-5alpha-delta24-nor-27-cholestene,3alpha,7alpha,12alpha-rihydroxy-5alpha-delta24-cholestene,5alpha-bufol and 5alpha-dermophol.