Determination of monoacetyldiamines and -polyamines in urine by high-performance liquid chromatography

Abstract

A procedure is described for the determination of monoacetylputrescine, N1-acetyl-spermidine and N8-acetylspermidine in human urine. The procedure is based on the high-performance liquid chromatographic separation of the 5-dimethylaminonaphthalene-1-sulfonyl (dansyl) derivatives of these amines using two different chromatographic modes. Monoacetyl-1,6-diaminohexane was used as an internal standard. The amines were extracted from urine using a silica gel cartridge. The dansyl monoacetylpolyamines were separated from the mixture of dansyl derivatives of urinary amines on a bonded-phase CN column using a programmed solvent gradient elution. The dansyl acetylpolyamines were rechromatographed on a silica gel column. This chromatographic procedure was used for the determination of the concentration of N1-acetylspermidine, N8-acetylspermidine and monoacetylputrescine in the urine of healthy volunteers and cancer patients.