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Chemistry
规模以上非靶向代谢组学剖析血清超高效液相色谱 - 质谱(UPLC-MS)
规模以上非靶向代谢组学剖析血清超高效液相色谱 - 质谱(UPLC-MS)
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Chemistry
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JoVE Journal Chemistry
Large Scale Non-targeted Metabolomic Profiling of Serum by Ultra Performance Liquid Chromatography-Mass Spectrometry (UPLC-MS)

规模以上非靶向代谢组学剖析血清超高效液相色谱 - 质谱(UPLC-MS)

Full Text
13,539 Views
07:34 min
March 14, 2013

DOI: 10.3791/50242-v

Corey D. Broeckling1, Adam L. Heuberger1, Jessica E. Prenni1

1Proteomics and Metabolomics Facility,Colorado State University

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Please note that some of the translations on this page are AI generated. Click here for the English version.

Overview

This article presents a protocol for non-targeted metabolite profiling of serum using ultra performance liquid chromatography coupled with mass spectrometry (UPLC-MS). It details the workflow from sample preparation to data analysis and metabolite identification.

Key Study Components

Area of Science

  • Metabolomics
  • Mass Spectrometry
  • Biochemistry

Background

  • Non-targeted metabolite profiling is crucial for understanding metabolic processes.
  • UPLC-MS is a widely used technique for analyzing complex biological samples.
  • Quality control is essential for ensuring the reliability of analytical results.
  • Identifying small molecule biomarkers can aid in various biomedical research applications.

Purpose of Study

  • To establish a robust method for metabolite extraction and analysis.
  • To ensure analytical stability through quality control measures.
  • To facilitate the identification of biomarkers in serum samples.

Methods Used

  • Extraction of metabolites in a plate format.
  • Establishment of a quality control sample.
  • Mass spectral data acquisition on QC and randomized serum samples.
  • Data pre-processing and statistical analysis for metabolite identification.

Main Results

  • Successful profiling of metabolites in serum samples.
  • Identification of potential small molecule biomarkers.
  • Demonstration of the robustness of the UPLC-MS method.
  • Insights into metabolic variations in serum.

Conclusions

  • The protocol provides a reliable framework for non-targeted metabolite profiling.
  • Quality control measures enhance the reproducibility of results.
  • UPLC-MS is effective for identifying biomarkers in serum.

Frequently Asked Questions

What is non-targeted metabolite profiling?
It is a technique used to analyze a wide range of metabolites in biological samples without prior knowledge of their identities.
Why is quality control important in metabolite profiling?
Quality control ensures the reliability and reproducibility of the analytical results over time.
What role does UPLC-MS play in this study?
UPLC-MS is used to separate and identify metabolites in serum samples effectively.
How can the results of this study be applied?
The results can be used to identify small molecule biomarkers for various diseases and conditions.
What are the key steps in the metabolite profiling workflow?
Key steps include sample preparation, data acquisition, data analysis, and metabolite identification.
Is this method applicable to other biological samples?
Yes, the method can be adapted for various biological matrices beyond serum.

超高效液相色谱-质谱联用 (UPLC-MS) 非靶向代谢物分析是研究代谢的一种强大技术。 本文概述了用于血清非靶向代谢物分析的典型工作流程,包括样品组织和制备、数据采集、数据分析、质量控制和代谢物鉴定。

该程序的总体目标是在大规模实验中对血清进行稳健的非靶向代谢物分析。首先,以板形式提取代谢物。建立稳定的 QC 样品和方法,以确保分析的长期稳定性。

然后重复采集 QC 和随机血清样品的质谱数据,进行数据、预处理、统计分析和代谢物鉴定。通过 U-P-L-C-M-S 进行非靶向代谢物分析的结果可用于鉴定血清中的小分子生物标志物。此处介绍了用于血清分析的方案。

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代谢组学分析 非靶向分析 UPLC-MS 血清 分析工作流程 样品制备 数据采集 质量控制 代谢物鉴定

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