Sensitive and selective analysis of coenzyme Q10 in human serum by negative APCI LC-MS
Sensitive and selective analysis of coenzyme Q10 in human serum by negative APCI LC-MS
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作者：
Hansen, G;Christensen, P;Lund, T
建立了两种新的高灵敏度、高选择性的LC-MS法测定人血清中总辅酶Q（10）浓度。该方法的灵敏度是基于CoQ（10）[M-]的自由基阴离子的非常有效的形成。负大气压电离，APCI（-）。[M-.]离子，m/z = 862.6，以选择性离子监测（SIM）或以MS 2模式（m/z = 862.6 -> m/z = 847.6）使用LCQ-十离子阱质谱仪进行。采用LC-APCI（2）-SIM和LC-APCI（2）-MS 2分析了两种标准血清样品（中等（0.73 μ g ml（-1））和高（1.96 μ g ml（-1））总CoQ（10）浓度），并将结果与电化学检测（ECD）的HPLC文献程序进行了比较。两种LC-MS方法均显示出更高的选择性，并且具有与HPLC-ECD方法相当或更好的灵敏度。中等浓度样品的LC-MS-SIM和LC-MS 2色谱图显示CoQ（10）信噪比分别为25和625。此外，还建立了一种简便、快速的血清前处理方法，该方法可在15 min内将血清中CoQ（10）H（2）定量氧化为CoQ（10）。
Two new sensitive and selective LC-MS methods have been developed for the quantification of the total coenzyme Q(10) concentration in human blood serum. The sensitivity of the methods is based on the very efficient formation of the radical anions of CoQ(10) [M-.] by negative atmospheric pressure ionisation, APCI(-). The mass detection of the [M-.] ions, m/z = 862.6, was performed either in selective ion monitoring (SIM) or in MS2 mode (m/z = 862.6 --> m/z = 847.6) using an LCQ-deca ion-trap mass spectrometer. Two standard serum samples with medium (0.73 mug ml(-1)) and high (1.96 mug ml(-1)) total CoQ(10) concentrations were analysed by LC-APCI(2)-SIM and LC-APCI(2)-MS2 and the results compared with a HPLC literature procedure with electrochemical detection (ECD). Both the LC-MS methods were shown to be more selective and with comparable or better sensitivity than the HPLC-ECD method. The LC-MS-SIM and LC-MS2 chromatograms of the medium concentration sample showed CoQ(10) signal to noise ratios of 25 and 625, respectively. In addition, a simple and fast serum pre-treatment procedure was developed, in which the serum CoQ(10)H(2) content was quantitatively oxidised quantitatively to CoQ(10) in less than 15 min by 1,4-benzoquinone.