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 chromatographic method, using pre-column derivatization with benzoyl chloride and ultraviolet detection, for the determination of N¹,N¹²-diacetylspermine in human urine. N¹,N¹²-diacetylspermine in the urine of 42 healthy subjects, 86 non-cancer patients, 11 pregnant women and 104 cancer patients (82 blood cancers and 22 solid cancers) were determined by our method. When 20-μ1 aliquots (equivalent to 50 μ1 of original urine) were injected into HPLC, N¹,N¹2-diacetylspermine was detected in only one (2.4%) of the healthy subjects and in 4(4.7%) of the non-cancer patients, but was detected in all of the pregnant women. In cancer patients, urinary N¹,N¹²-diacetuylspermine was detected in 39 cases (47.6%) of blood cancer and in 8 cases (36.4%) of solid cancer. In addition, we also measured urinary N¹,N⁸-diacetylspermidine in a proportion of the healthy subjects and cancer patients using our HPLC method with minor modifications. Finally, free, monoacetylated and diacetylated polyamines were determined in the urine of a woman during pregnancy, and our results were consistent with those reported by Van den Berg et al. (1986).