Determination of polyamines in urine

Abstract

A method is described for the determination of putrescine, cadaverine, spermidine and spermine in urine. The procedure involves hydrolysis of 1 ml of urine in 6 N HCl at 120° overnight, adjustment to pH 13, and extraction into isoamyl alcohol. The evaporated solution is treated with dansyl chloride, and the dansyl polyamines are chromatographed on a thin-layer plate of alumina G (Woelm). A solvent of dioxane-chloroform (1:200) allows good separation of putrescine, cadaverine and spermidine, but spermine is not always distinguishable. Dioxane-acetic acid-chloroform (2:1:97) separates cadaverine, spermidine and spermine, but putrescine is not well separated from ammonia. Preliminary treatment of urine with urease should be effected if it is required to eliminate ammonia. Dansyl polyamines are measured fluorimetrically after elution. Normal values are given for each polyamine. Elevated amounts of spermidine, and sometimes of spermine as well, are frequently found in cancer patients. Raised excretions of putrescine and cadaverine are occasionally encountered, the latter possibly due to bacterial action.