High-Speed Quantitative UPLC-MS Analysis of Multiple Amines in Human Plasma and Serum via Precolumn Derivatization with 6-Aminoquinolyl-N-hydroxysuccinimidyl Carbamate: Application to Acetaminophen-Induced Liver Failure
High-Speed Quantitative UPLC-MS Analysis of Multiple Amines in Human Plasma and Serum via Precolumn Derivatization with 6-Aminoquinolyl-N-hydroxysuccinimidyl Carbamate: Application to Acetaminophen-Induced Liver Failure
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DOI：
10.1021/acs.analchem.6b04623
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发表时间：
2017-02-21
影响因子：
7.4
通讯作者：
Nicholson, Jeremy K.
中科院分区：
文献类型：
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作者：
Gray, Nicola;Zia, Rabiya;Nicholson, Jeremy K.
开发了一种靶向反相梯度UPLC-MS/MS测定法，用于使用6氨基喹啉-N-羟基琥珀酰亚胺氨基甲酸酯（AccQTag Ultra）进行柱前衍生化，定量/监测人血浆和血清中的66种氨基酸和含氨基化合物。目标胺的衍生化需要最少的样品制备，并导致具有优异的色谱和质谱检测性能的分析物。所得到的方法，它只需要10 μ L的样品，提供了66个分析物在7.5分钟内，包括基线分辨率的异构体，如亮氨酸和异亮氨酸的可重复性和强大的分离。该测定法已被验证的33个氨基化合物（主要是氨基酸）的定量浓度范围从2到20和800 μ M。日内和日间准确度分别为0.05 - 15.6%和0.78-13.7%，精密度分别为0.91 - 16.9%和2.12-15.9%。另外33种生物胺可以在样品中监测浓度的相对变化，而不是定量。将该测定应用于来自健康对照和患有对乙酰氨基酚（APAP，扑热息痛）诱导的急性肝衰竭（ALF）的患者的样品，显示各组之间芳香族和支链氨基酸以及许多其他分析物的量存在显著差异，包括ALF患者中肌氨酸浓度增加的新观察结果。所开发的检测方法的特性（包括分析时间短）使其适用于临床和流行病学环境中的高通量靶向UPLC-ESI-MS/MS代谢组学分析。
A targeted reversed-phase gradient UPLC-MS/MS assay has been developed for the quantification /monitoring of 66 amino acids and amino-containing compounds in human plasma and serum using precolumn derivatization with 6aminoquinolyl-N-hydroxysuccinimidyl carbamate (AccQTag Ultra). Derivatization of the target amines required minimal sample preparation and resulted in analytes with excellent chromatographic and mass spectrometric detection properties. The resulting method, which requires only 10 mu L of sample, provides the reproducible and robust separation of 66 analytes in 7.5 min, including baseline resolution of isomers such as leucine and isoleucine. The assay has been validated for the quantification of 33 amino compounds (predominantly amino acids) over a concentration range from 2 to 20 and 800 mu M. Intra-and interday accuracy of between 0.05 and 15.6 and 0.78-13.7% and precision between 0.91 and 16.9% and 2.12-15.9% were obtained. A further 33 biogenic amines can be monitored in samples for relative changes in concentration rather than quantification. Application of the assay to samples derived from healthy controls and patients suffering from acetaminophen (APAP, paracetamol)-induced acute liver failure (ALF) showed significant differences in the amounts of aromatic and branched chain amino acids between the groups as well as a number of other analytes, including the novel observation of increased concentrations of sarcosine in ALF patients. The properties of the developed assay, including short analysis time, make it suitable for high-throughput targeted UPLC-ESI-MS/MS metabonomic analysis in clinical and epidemiological environments.