Determination of Free and Total Polyamines in Human Serum and Urine by Ion-pairing High-performance Liquid Chromatography Using a Radial Compression Module. Application to Blood Polyamine Determination in Cancer Patients Treated or Not Treated With...

A sensitive and rapid determination of free and total polyamines (putrescine, cadaverine, spermidine and spermine) in urine and serum is described. The procedure is based on reversed-phase high-performance liquid chromatographic separation using radial compression module (Radialpak C 8). The samples are purified with a silica gel Sep-Pak cartridge. The polyamines are converted to dansyl chloride derivatives and separated using a linear gradient of triethylammonium phosphate-methanol within 15 min. The lower limits of detection are 10 pmoles for spermine and 5 pmoles for other polyamines. This method is applied to cancer patients treated by cytotoxic chemotherapy with or without difluoromethylornithine (DFMO). All four polyamines are significantly increased in these patients before treatment. On day 8, after onset of treatment, the levels of polyamines in patients not treated with DFMO are more elevated than on day 1, while in patients treated with DFMO the levels are decreased. However, DFMO does not seem to modify the treatment result. The patients which have a low level of putrescine before and during treatment, do not respond to treatment. Perhaps this low level is the consequence of conjugation of this polyamine?