A rapid chemiluminescence-based method for the determination of total polyamines in biological samples

Abstract

The metabolism of the natural polyamines putrescine, spermidine and spermine is closely associated with cell proliferation [l]. Although in human cancer, the concentrations of polyamines in serum and urine are mostly higher than in healthy persons, the determination of extracellular polyamines obviously does not represent a reliable method for early detection of cancer (see [2]). However, an acute rise in serum polyamine levels or in their urinary excretion in response to successful anti-cancer therapy [3], as the result of the release from damaged cells of these strictly intracellular compounds, may be helpful for the evaluation of the efficacy of a given anti-cancer regimen. In addition, serial determinations of extracellular polyamines have a distinct value in the surveillance of tumor regression or relapse in certain tumor types, especially in medulloblastomas, where an increase in putrescine in cerebrospinal fluid predicts the regrowth of the tumor well before it is detected by any other diagnostic techniques [4]. A number of sensitive methods are available for determination of polyamines in biological samples including extracellular fluids. Dansylation of polyamines followed by chromatography on thin layer plates [S] has the drawback of being time-consuming. High performance liquid chromatography [6] and liquid ion exchange chromatography [7] are sensitive methods but somewhat cumbersome for efficient handling of large numbers of samples. Radioimmunological methods utilizing antibodies to the polyamines [8] are useful for rapid analyses, but require a good quality antibody. We have now developed a rapid (up to 20 samples/h) and sensitive (in the pmol range) method for the determination of total polyamines (putrescine + spermidine +