Shimba Kawasue, Yohei Sakaguchi, Ena Yano, Tadashi Hayama, Reiko Koga, H. Yoshida, H. Nohta
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引用次数: 1
摘要
姜黄素已被证明在预防和治疗各种人类疾病方面具有药理活性。在本研究中,我们开发了一种用液相色谱(LC)串联质谱(MS/MS)选择性分析姜黄素的荧光衍生方法。姜黄素与含硫醇的含氟试剂(1H,1H,2H,2H-全氟-1-癸硫醇)在温和条件下通过Michael加成反应进行衍生,并将获得的衍生物引入含氟LC柱(Fluofix II 120E,150×2.0mm i.d.,5μm,Wako)。柱上选择性保留的氟衍生姜黄素也能够通过负电喷雾电离MS/MS进行高灵敏度检测。人血清样品的预处理采用CH3CN蛋白沉淀法。本方法得到的校准曲线在7.4-442 ng/mL血清范围内呈良好的线性关系(r2=0.9998),检测限(S/N=3)和定量限(S/N=10)分别为1.8 ng/mL血清和6.1 ng/mL血清。本方法已成功应用于人血清样品中姜黄素的分析。
Fluorous Derivatization Method for Selective Analysis of Curcumin with Liquid Chromatography-Tandem Mass Spectrometry
Curcumin has been shown to be pharmacologically active in the prevention and treatment of various human diseases. In this study, we developed a fluorous derivatization method for selective analysis of curcumin with liquid chromatography (LC)tandem mass spectrometry (MS/MS). Curcumin was derivatized with the thiol-containing fluorous reagent (1H,1H,2H,2Hperfluoro-1-decanethiol) under mild conditions via Michael addition reaction, and the obtained derivative was introduced to a fluorous LC column (Fluofix-II 120E, 150 × 2.0 mm i.d., 5 μm, Wako). The selectively retained fluorous-derivatized curcumin on the column was also enabled highly sensitive detection with negative electrospray ionization MS/MS. Pretreatment of human serum sample was used protein precipitation with CH3CN. The calibration curve obtained by the present method showed good linearity (r2 = 0.9998) in the range of 7.4-442 ng/mL serum, and the limit of detection (S/N = 3) and limit of quantification (S/N = 10) were 1.8 ng/mL serum and 6.1 ng/mL serum, respectively. The present method was successfully applied to the analysis of curcumin in human serum sample.