Detecting low-abundance vasoactive peptides in plasma: Progress toward absolute quantitation using nano liquid chromatography-mass spectrometry
Detecting low-abundance vasoactive peptides in plasma: Progress toward absolute quantitation using nano liquid chromatography-mass spectrometry
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DOI：
10.1016/j.ab.2009.07.021
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发表时间：
2009-11-15
影响因子：
2.9
通讯作者：
Hook, Vivian
中科院分区：
文献类型：
--
作者：
Lortie, Mark;Bark, Steven;Hook, Vivian
分析与功能相关的多肽激素浓度的变化对于了解许多疾病和治疗的病因至关重要。我们提出了使用纳米液质联用(LC-MS)同时检测50亩L血浆样本中选定的一组血管活性多肽(血管紧张素、缓激肽和相关激素)的新数据，使啮齿动物模型能够重复采样。通过对目标多肽的层析和多反应监测来提高MS的灵敏度，获得了低至10(-17)摩尔的线性响应。通过甲醇沉淀或离线高效液相色谱(HPLC)分离纯化血浆多肽，能够检测内源性多肽，并揭示了提高回收率的方法。作为原理的证明，在麻醉大鼠的急性血管紧张素转换酶(ACE)抑制之前、期间和之后，对七种血管活性多肽进行了分析。值得注意的是，血管舒张性缓激肽在药物注射后明显增加10倍，但血管紧张素II水平变化相对较小。用于描述与功能相关的多肽或其他分析物的靶向MS分析将极大地增强我们定义调节复杂和动态生理过程的事件序列的能力。(C)2009 Elsevier Inc.保留所有权利。
Profiling changes in the concentration of functionally related peptide hormones is critical to understanding the etiology of many diseases and therapies. We present novel data using nano liquid chromatography-mass spectrometry (LC-MS) to simultaneously measure a select group of vasoactive peptides (angiotensin, bradykinin, and related hormones) in 50-mu l plasma samples, enabling repeated sampling in rodent models. By chromatographically resolving target peptides and using multiple reaction monitoring to enhance MS sensitivity, linear responses down to 10(-17) mol were achieved. Purification of plasma peptides by either methanol precipitation or off-line high-performance liquid chromatography (HPLC) fractionation enabled the detection of endogenous peptides and revealed approaches for enhancing recovery. As proof of principle, seven vasoactive peptides were profiled before, during, and after acute angiotensin-converting enzyme (ACE) inhibition in an anesthetized rat. Of note was an apparent 10-fold increase in vasodilatory bradykinin that reversed after drug infusion but relatively minor changes in angiotensin II levels. Targeted MS analysis used to profile functionally related peptides or other analytes will greatly enhance our ability to define the sequence of events regulating complex and dynamic physiological processes. (C) 2009 Elsevier Inc. All rights reserved.