抄録
An improved assay method for urinary polyamines using an automated amino acid analyzer was developed. Three ml of human urine was hydrolyzed in 6N HC1 at 110°C for 24 hrs. 1, 6-Diaminohexane (200 n moles) was added to serve as internal standard.The hydrolysates were evaporated to dryness at 60°C and then 3.5 ml of 50 mM EDTA (pH 7.0) was added. The supernatants were applied on the column (9×40mm) of Dowex 50W-X8 (H+) and amino acids were washed out with sodium phosphate buffer and HCl after the method of Inoue and Mizutani (1973). The polyamines were eluted with 6N HCl. The eluates were evaporated to dryness and then 0.5 ml of distilled water was added. The polyamines in aqueous extracts were analyzed by an automated amino acid analyzer. Amino acids and other ninhydrin-positive contaminants were first removed by the citrate buffer (pH 5.25) containing 0.5 M KCl. Polyamines and some biogenic amines were then eluted by the citrate buffer (pH 5.25) containing 1.6 M KCl.
Putrescine, spermidine, 1, 3-diaminopropane and cadaverine were found in all the urine samples analyzed. Spermine and histamine was detected only in some normal subjects and cancer patients. Urinary excretion of putrescine and spermidine in terms of μ moles per gm. creatinine was significantly increased in patients with advanced breast cancer. A ninhydrin-positive peak which eluted just before cadaverine was observed in some patients with breast cancer. The diagnostic significance of this unknown substance remains to be elucidated.
