High-performance liquid chromatographic determination of diacetylpolyamines using benzoyl chloride as a derivatizing agent: application to human urine

We developed a simple reversed-phase high-performance liquid chromatographi c method, using pre-column derivatization with benzoyl chloride and ultravi olet detection, for the determination of N-1,N-12-diacetylspermine in human urine. N-1,N-12-diacetylspermine in the urine of 42 healthy subjects, 86 n on-cancer patients, 11 pregnant women and 104 cancer patients ( 82 blood ca ncers and 22 solid cancers) were determined by our Method. When 20-mul aliq uots (equivalent to 50 mul of original urine) were injected into HPLC, N-1, N-12-diacetylspermine was detected in only one (2.4%) of the healthy subjec ts and in 4 (4.7%) of the non-cancer patients, but was detected in all of t he pregnant women. In cancer patients, urinary N-1,N-12-diacetylspermine wa s detected in 39 cases (47.6%) of blood cancer and in 8 cases (36.4%) of so lid cancer. In addition, we also measured urinary N-1,N-8-diacetylspermidin e in a proportion of the healthy subjects and cancer patients using our HPL C method with minor modifications. Finally, free. monoacetylated and diacet ylated polyamines were determined in the urine of a woman during pregnancy, and our results were consistent with those reported by Van den Berg et at. (1986).