Determination of free polyamines in human bile by high-performance liquid chromatography

1. Polyamines are widely distributed in the body and may have cholesterol-nucleating activity in model bile and human bile. There are only partial and scant data available on the type of polyamines in human bile. 2. In this study methods for extraction of free polyamines, benzoylation and an h.p.l.c.-based method for the quantitative determination of putrescine, cadaverine, spermidine and spermine in bile are described. The h.p.l.c. methodology was validated and compared with separation on t.l.c. after dansylation. 3. The polyamine content of 11 gallbladder bile samples and 11 T-tube bile samples was analysed, all from patients with gallstones. Polyamines were found in three out of 11 gallbladder bile samples and eight out of 11 T-tube bile samples. Putrescine levels were 0.5-287 mumol/l and cadaverine levels were 2.4-645.4 mumol/l; these were considerably higher than spermine and spermidine levels. 4. As many of these gallstones bile samples were devoid of polyamines, it is questionable whether polyamines play an important role in cholesterol nucleation in human bile.