在数据依赖性获取中对质谱参数的优化,用于基于假定赋值的非目标代谢学
Hailemariam Abrha Assress1,2, Mario G Ferruzzi1,2, Renny S Lan1,2
1Arkansas Children's Nutrition Center, Little Rock, Arkansas 72202, United States.
概括
优化质谱学参数,如MS/MS事件和信号值,显著提高了在非目标代谢学中的代谢物识别. 本研究提供了改善复杂生物样本中代谢物覆盖率和光谱质量的策略.
科学领域:
- 分析化学 分析化学
- 生物化学 生化学
- 质谱测量质量谱测量
背景情况:
- 非定位代谢学依赖于全面的代谢物识别.
- 数据依赖采集 (DDA) 质谱需要仔细的参数优化,以获得最大的MS/MS覆盖范围.
- 准确的代谢物注释对于理解生物途径至关重要.
研究的目的:
- 研究各种质谱参数对非目标代谢学中的代谢物注释的影响.
- 确定Exploris 480-Orbitrap质谱仪的最佳设置,以改善代谢物识别.
- 提出提高代谢物覆盖率和光谱质量的策略.
主要方法:
- 系统地探索质谱参数,包括质分辨率,射频水平,信号强度值,MS/MS事件数量,周期时间,碰撞能量,最大离子注入时间 (MIT),动态排除和自动增强控制 (AGC) 目标值.
- 在数据依赖采集 (DDA) 实验中使用了Exploris 480-Orbitrap质谱仪.
- 评估了参数变化对代谢物注释率的影响.
主要成果:
- 通过十次MS/MS扫描,2.0m/z隔离窗口,1x10^4信号强度值,180,000个MS分辨率和30,000个MS/MS分辨率在70%的射频水平上,实现了最佳的注释.
- 将5x10^6 (MS) 和1x10^5 (MS/MS) 的AGC目标值与100 ms (MS) 和50 ms (MS/MS) 的MIT相结合,改善了代谢物注释.
- 一个10秒的排除时间和两阶段碰撞能量优化了光谱质量.
结论:
- 质谱参数显著影响代谢学结果,影响代谢物覆盖率和识别信心.
- 该研究为DDA实验提供了优化的参数,以增强在非目标代谢学中的代谢物识别.
- 建议使用真实标准进行进一步验证,并在不同的协议上进行测试.
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