Hishinuma Takanori, Suzuki Kaori, Saito Masayoshi, Yamaguchi Hiroaki, Suzuki Naoto, Tomioka Yoshihisa, Kaneko Izumi, Ono Masao, Goto Junichi
Graduate School of Pharmaceutical Sciences, Tohoku University, and Department of Pharmaceutical Sciences, Tohoku University Hospital, Sendai, Japan.
Prostaglandins Leukot Essent Fatty Acids. 2007 Jun;76(6):321-9. doi: 10.1016/j.plefa.2007.04.005. Epub 2007 Jun 21.
We have developed a method for the simultaneous estimation of the levels of the prostanoids 6-keto prostaglandin (PG) Flalpha, PGB2, PGD2, PGE2, PGF2(alpha), PGJ2, and thromboxane (TX) B2 in blood- or serum-containing medium using liquid chromatography-tandem mass spectrometry. These prostanoids and their deuterium derivatives, which were used as internal standards, were subjected to solid-phase extraction using Empore C18 HD disk cartridges and analyzed in the selected reaction-monitoring mode. A linear response curve starting at 10 pg of prostanoid/tube was observed for each prostanoid. The accuracy of the method was demonstrated with samples containing known amounts of the prostanoids. Furthermore, we used this method to analyze the prostanoids produced in mouse bone marrow-derived mast cells stimulated with arachidonic acid, which resulted in the production of PGD2, PGE2, PGF2alpha, and TXB2. The results suggest that this simultaneous quantification method is useful for the analysis of the production of biomedically important prostanoids.
我们开发了一种使用液相色谱-串联质谱法同时测定含血或血清培养基中前列腺素6-酮前列腺素（PG）F1α、PGB2、PGD2、PGE2、PGF2α、PGJ2和血栓素（TX）B2水平的方法。这些前列腺素及其用作内标的氘代衍生物，使用Empore C18 HD盘式柱进行固相萃取，并在选择反应监测模式下进行分析。每种前列腺素均观察到从10 pg前列腺素/管开始的线性响应曲线。该方法的准确性通过含有已知量前列腺素的样品得到证明。此外，我们使用该方法分析了用花生四烯酸刺激的小鼠骨髓来源肥大细胞中产生的前列腺素，结果产生了PGD2、PGE2、PGF2α和TXB2。结果表明，这种同时定量方法对于分析具有重要生物医学意义的前列腺素的产生是有用的。
