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Mass Spectrometry: Carboxylic Acid, Ester, and Amide Fragmentation01:01

Mass Spectrometry: Carboxylic Acid, Ester, and Amide Fragmentation

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The fragmentation patterns observed for compounds such as carboxylic acids, esters, and amides in the mass spectra include ⍺-cleavage and McLafferty rearrangement. Fragmentation by ⍺-cleavage preferentially occurs at the carbon-carbon bond at the ⍺-position next to the carboxylic group to generate a neutral radical and a cation. Long chain compounds with hydrogen at their γ-carbon undergo McLafferty rearrangement to give a radical cation and a neutral alkene.
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Mass Spectrometry: Overview01:19

Mass Spectrometry: Overview

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Mass spectrometry is an analytical technique used to determine the molecular mass and molecular formula of a compound. The basic principle of mass spectrometry is to generate ions from the analyte molecule and measure these ion abundances against their molecular mass.  One common type of ionization, known as electrospray ionization or EI, bombards the analyte molecules in the gas phase with high-energy electron beams. The electron beams displace an electron from the molecule and leave...
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Mass Spectrometry: Complex Analysis01:21

Mass Spectrometry: Complex Analysis

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Mass spectrometry is an important technique for the identification of pure compounds. However, it has some limitations for the analysis of complex mixtures, often due to excessive fragmentation making the spectrum too complicated to decipher. Mass spectrometry can be combined with suitable separation methods in sequence, forming hyphenated methods, which are useful in the analysis of complex mixtures.
GC–MS is a powerful hyphenated method commonly used in forensics and environmental...
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Mass Spectrometry: Amine Fragmentation00:55

Mass Spectrometry: Amine Fragmentation

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Amines can be identified using mass spectroscopy based on their characteristic fragmentation patterns. The molecular ions of amines undergo fragmentation via ⍺-cleavage. The ⍺-cleavage of the carbon-carbon bonds in amines generates an alkyl radical and resonance-stabilized nitrogen-containing cation.
In amines, the number of nitrogen atoms affects the mass of the molecular ion, which is described by the nitrogen rule of mass spectrometry. This rule states that a compound containing...
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In mass spectroscopy, amines undergo fragmentation to give parent ions with odd molecule weights. This observed mass spectrum follows the nitrogen rule: a molecule with an odd number of nitrogen atoms produces a parent ion with an odd molecular weight. The remaining fragments have an even mass.
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Mass Spectrometers

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This lesson details the instrumentation of a mass spectrometer—a physical instrument to perform mass spectrometry on analyte molecules and record the characteristic mass spectra. This is achieved via three chief functions:
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缓冲器4-乙基/乙酸盐:探索原生质谱学的新替代缓冲器.

Darya Hadavi1, Che Yee Ng1, Yuandi Zhao1

  • 1Maastricht Multi Modal Molecular Imaging (M4i) Institute, Division of Imaging Mass Spectrometry (IMS), Maastricht University, Maastricht, The Netherlands.

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概括
此摘要是机器生成的。

一种新的4-乙烯基/乙酸盐 (4EM/A) 缓冲剂在原生质谱 (MS) 研究中保留了蛋白质结构和生物活性. 这种缓冲区提供了增强的结构稳定性,并使得分析具有挑战性的蛋白质,如心脏托罗宁T.

关键词:
收费状态 分配状态 分配乙基摩尔福林 (Ethylmorpholine) 是一种有机物.离子流动性光谱仪离子流动性光谱仪原生质谱法本土质谱法.蛋白质复合体 蛋白质复合体

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科学领域:

  • 生物物理化学 生物物理化学
  • 分析化学 分析化学
  • 结构生物学 结构生物学

背景情况:

  • 原生质谱 (MS) 需要挥发性,电喷离子化 (ESI) 兼容的缓冲器.
  • 传统的缓冲剂,如酸 (AA),可以在ESI-MS过程中诱导蛋白质展开和酸化.
  • 需要一个缓冲区来维持蛋白质的原始状态,并模拟生理条件.

研究的目的:

  • 为了评估本地MS分析的4-乙烯基/乙酸盐 (4EM/A) 缓冲区.
  • 为了比较4EM/A与酸 (AA) 蛋白质和蛋白质复合体分析.
  • 评估4EM/A对蛋白质生物活性和结构完整性的影响.

主要方法:

  • 将4EM/A缓冲区与AA缓冲区进行蛋白质 (5103 kDa) 和蛋白质复合物的比较.
  • 分析了原生MS配置文件,碰撞截面 (CCS) 值和到达时间分布 (ATD).
  • 评估了蛋白质的生物活性,并分析了具有挑战性的蛋白质,如人类心脏热素复合物 (cTn复合物).

主要成果:

  • 4EM/A导致较低的蛋白质电荷状态,在纳米-ESI和MS期间保持本地折叠.
  • 离子流动性在4EM/A中显示出减少的构造变化,表明增强的稳定性和紧性.
  • 莱索酶在4EM/A中表现出增加的生物活性;4EM/A首次启用了心脏特罗邦尼T (cTnT) 的本地MS分析.

结论:

  • 引入了4EM/A (pKa 7.72/4.76) 作为本地MS.的优越缓冲器.
  • 4EM/A保持了蛋白质和蛋白质复合物的生物活性和结构完整性.
  • 4EM/A是高级本地MS应用程序的一个有前途的工具.