Ducros Véronique, Ruffieux Daniel, Belva-Besnet Hélène, de Fraipont Florence, Berger François, Favier Alain
Département de Biologie Intégrée, Centre Hospitalier Universitaire, 38043 Grenoble cedex, France.
Anal Biochem. 2009 Jul 1;390(1):46-51. doi: 10.1016/j.ab.2009.04.007. Epub 2009 Apr 11.
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.
红细胞（RBC）中多胺的浓度被认为是细胞增殖的一个指标。已证明该指标对某些癌症患者的随访和治疗具有临床重要性。红细胞中多胺的浓度通常通过高效液相色谱（HPLC）结合荧光检测来测定。在当前工作中，我们提出了一种液相色谱 - 串联质谱（LC - MS/MS）方法，用于定量红细胞中的三种主要多胺——腐胺、亚精胺和精胺。多胺经丹磺酰化后，在C18柱上通过20分钟的LC梯度进行分析，并与串联质谱仪在线联用。使用内标（1,8 - 二氨基辛烷）进行定量。该方法对三种多胺表现出优异的线性，回归系数高于0.99。腐胺、亚精胺和精胺的检测限分别为0.10、0.75和0.50 pmol/ml。腐胺、亚精胺和精胺的批内精密度值均优于10%，批间精密度值分别为13%、9%和20%。LC - MS/MS方法足够简单且可靠，足以取代目前使用的HPLC荧光检测方法，在该方法中腐胺并非总能被检测到。
