Determination of dansylated polyamines in red blood cells by liquid chromatography-tandem mass spectrometry.

The concentration of polyamines in red blood cells (RBCs) is considered to be an index of cell proliferation. This index has been demonstrated to be of clinical importance for the follow-up and treatment of some cancer patients. The concentration of polyamines in RBCs is usually determined by high-performance liquid chromatography (HPLC) with fluorescence detection. In the current work, we present a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the quantification of putrescine, spermidine, and spermine, the three major polyamines in RBCs. The polyamines were dansylated and analyzed by an LC gradient of 20-min duration on a C18 column on-line with a tandem mass spectrometer. An internal standard (1,8-diaminooctane) was used for quantification. This method exhibited excellent linearity for the three polyamines with regression coefficients higher than 0.99. The limits of detection for putrescine, spermidine, and spermine were 0.10, 0.75, and 0.50 pmol/ml, respectively. The intrarun precision values for putrescine, spermidine, and spermine all were better than 10%, and the interrun precision values were 13%, 9%, and 20%, respectively. The LC-MS/MS method is sufficiently simple and reliable enough to replace the currently used HPLC method with fluorescence detection in which putrescine is not always detectable.

Relationships between selenium, lipids, iron status and hormonal therapy in women of the SU.VI.M.AX cohort.

Significant differences in serum selenium concentration according to contraceptive treatment and age have been evidenced in women of the SU.VI.M.AX cohort. This study aimed at verifying the physiopathological hypothesis that the observed increase in serum selenium concentration could be related to serum lipid increase and/or bleeding decrease. Women were divided into six groups: menopausal with or without hormonal replacement therapy; non-menopausal using contraceptive pills; intrauterine device; other contraceptive treatment or no contraceptive treatment. Adjusted linear regression indicated positive associations between selenium and apolipoprotein A1 (r(2) from 0.038 to 0.074, p<0.07 depending on groups) or ferritin in serum (r(2) from 0.032 to 0.075, p<0.07 depending on groups). These relationships could explain the differences observed according to hormonal treatment and age in the SU.VI.MAX study.