Simultaneous quantification of S-adenosyl methionine and S-adenosyl homocysteine in human plasma by stable-isotope dilution ultra performance liquid chromatography tandem mass spectrometry
Simultaneous quantification of S-adenosyl methionine and S-adenosyl homocysteine in human plasma by stable-isotope dilution ultra performance liquid chromatography tandem mass spectrometry
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DOI：
10.1016/j.jchromb.2009.09.039
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发表时间：
2009-11-15
影响因子：
3
通讯作者：
Obeid, Rima
中科院分区：
文献类型：
--
作者：
Kirsch, Susanne H.;Knapp, Jean-Pierre;Obeid, Rima
S-腺苷蛋氨酸是由蛋氨酸形成的一种重要的甲基供体。S-腺苷同型半胱氨酸是腺苷同型半胱氨酸去甲基化后形成的，是许多甲基转移酶的有效抑制剂。建立了一种改进的稳定同位素稀释超高效液相色谱串联质谱(UPLC-MS/MS)同时测定生物样品中SAM和SAH的方法。该方法包括含有苯基硼酸的固相萃取过程，用于SAM和SAH的结合和净化。样品提取后，使用高效液相色谱柱SymmetryShield RP18或Acquity UPLC BEH C-18色谱柱与高效液相-MS/MS或UPLC-MS/MS系统分离和检测。采用Acquity UPLC BEH C-18色谱柱检测效果最佳。在血浆样品中，SAM和SAH的批内变异系数(CVs)分别为3.3%和3.9%，批间变异系数分别为10.1%和8.3%。两种不同浓度下的平均回收率分别为100.0%和101.7%。SAM和SAH的定量限分别为0.5nmol/L和0.7nmol/L。31份血浆中SAM和SAH的平均浓度分别为85.5(11.1)nmol/L和13.3(5.0)nmol/L，SAM/SAH比值为7.0(1.8)。新的UPLC-MS/MS方法对SAM和SAH具有很高的灵敏度和选择性，变异系数低，样品制备速度快(60min内40个样品)，分析时间(3min)。这种新的检测方法可以用于大规模的临床研究。(C)2009爱思唯尔B.V.保留所有权利。
S-adenosyl methionine (SAM) is an important methyl group donor that is formed from methionine. S-adenosyl homocysteine (SAH) is formed after demethylation of SAM and represents a potent inhibitor of many methyltransferases. We developed an improved stable-isotope dilution ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method for the simultaneous quantification of SAM and SAH in biological samples. The method comprises a phenylboronic acid-containing solid-phase extraction procedure, serving for binding and clean-up of SAM and SAH. After extraction, samples were separated and detected using either a HPLC SymmetryShield RP18 or an Acquity UPLC BEH C-18 column with a HPLC-MS/MS or an UPLC-MS/MS system. The best results were obtained by Acquity UPLC BEH C-18 column. In plasma samples, the estimated intraassay coefficients of variation (CVs) for SAM and SAH were 3.3% and 3.9%, respectively, the interassay CVs were 10.1% for SAM and 8.3% for SAH. Mean recovery of SAM and SAH at two different concentrations was 100.0% for SAM and 101.7% for SAH. The quantification limits were 0.5 and 0.7 nmol/L for SAM and SAH, respectively. In 31 plasma samples, the mean concentrations (SD) were 85.5 (11.1) nmol/L for SAM and 13.3 (5.0) nmol/L for SAH with a SAM/SAH ratio of 7.0 (1.8). The new UPLC-MS/MS method showed very high sensitivity and selectivity for SAM and SAH, low CVs and fast sample preparation (40 samples in 60 min) and analysis time (3 min). This new assay can be used for large-scale clinical studies. (C) 2009 Elsevier B.V. All rights reserved.