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首页> 美国卫生研究院文献>other >An Interface for Sensitive Analysis of Monoamine Neurotransmitters by Ion-Pair Chromatography–ESI-MS with Continuous On-Line Elimination of Ion-Pair Reagents
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An Interface for Sensitive Analysis of Monoamine Neurotransmitters by Ion-Pair Chromatography–ESI-MS with Continuous On-Line Elimination of Ion-Pair Reagents

机译：离子对色谱-ESI-MS对单胺类神经递质进行灵敏分析的接口可连续在线消除离子对试剂

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A challenge in coupling ion-pair chromatography (IPC) on-line with electrospray ionization-mass spectrometry (ESI-MS) is that the nonvolatile ion-pair reagent (e.g., alkyl sulfate for amines or tetrabutylammonium for carboxylic acids) in the mobile phase suppresses the ESI-MS signals in the gas phase and their accumulation can clog the MS sampling interface. Consequently, IPC–ESI-MS is conducted either with a volatile ion-pair reagent, which could compromise the analyte separation efficiency, or with a downstream ion-exchange column to rid the ion-pair reagents of the mobile phase. In the latter approach, the limited capacity of ion-exchange columns requires frequent off-line column regeneration, which affects the separation throughput and prohibits long separations from being performed. A dual-valve, dual-ion exchange column interface of IPC–ESI-MS is designed for undisrupted separations and simultaneous column regeneration. Owing to the efficacy in removing the ion-pair reagent, the detection of eluents of monoamine neurotransmitters by an ion trap MS results in the limits of detection of 0.03 μM for dopamine or DA and 0.01 μM for 5-hydroxytryptamine or 5-HT. These values are lower than those obtained with ion trap MS of similar sensitivity when combined with the use of specialized chromatographic columns or sample preconcentration. Excellent reproducibility was attained with repeatedly regenerated ion-exchange columns (RSD = 4–6%) for an extended period of time (RSD < 6% for six days). DA and 5-HT in rat straital extracts were analyzed and our data demonstrate that interferences inherent in the tissues and the ion-pair reagent have been successfully eliminated. This simple interface should be readily amenable to the separation and MS analysis of other types of polar compounds in complex sample media.
机译：在线结合离子对色谱法（IPC）与电喷雾电离质谱（ESI-MS）的挑战在于，流动相中的非挥发性离子对试剂（例如，胺的烷基硫酸盐或羧酸的四丁基铵）抑制气相中的ESI-MS信号，其累积会阻塞MS采样界面。因此，IPC–ESI-MS可以使用挥发性离子对试剂（可能会影响分析物的分离效率）进行操作，也可以使用下游离子交换柱进行离子交换，以去除流动相中的离子对试剂。在后一种方法中，有限的离子交换柱容量需要频繁的离线柱再生，这会影响分离通量并阻碍长距离分离的进行。 IPC–ESI-MS的双阀双离子交换柱接口设计用于不间断的分离和同时的柱再生。由于去除离子对试剂的功效，通过离子阱质谱仪检测单胺类神经递质的洗脱液导致多巴胺或DA的检出限为0.03μM，5-羟基色胺或5-HT的检出限为0.01μM。当与专用色谱柱或样品预浓缩结合使用时，这些值低于使用具有相似灵敏度的离子阱质谱仪获得的值。反复再生的离子交换柱（RSD = 4–6％）延长了一段时间（RSD <6％，持续6天），可获得出色的重现性。分析了大鼠纹状体提取物中的DA和5-HT，我们的数据表明已成功消除了组织和离子对试剂中固有的干扰。这种简单的界面应易于适应复杂样品介质中其他类型极性化合物的分离和质谱分析。

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