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相关概念视频

High-Performance Liquid Chromatography: Introduction01:11

High-Performance Liquid Chromatography: Introduction

High-performance liquid chromatography(HPLC), formerly referred to as High-pressure liquid chromatography, is a powerful technique used to separate, identify, and quantify components in complex mixtures. The term "high pressure" refers to using high pressure to push the liquid mobile phase through the tightly packed columns.
In HPLC, two phases play a critical role in the separation process:
High-Performance Liquid Chromatography: Elution Process01:05

High-Performance Liquid Chromatography: Elution Process

In High-Performance Liquid Chromatography (HPLC), the elution process is critical to the separation of analytes and the quality of chromatographic results. Elution describes how compounds move through the column and separate based on their interactions with the mobile and stationary phases. This process determines the resolution, peak shape, and retention times in the chromatogram, which are essential for identifying and quantifying components in complex mixtures. Understanding the elution...
High-Performance Liquid Chromatography: Instrumentation00:57

High-Performance Liquid Chromatography: Instrumentation

High-performance liquid chromatography, or HPLC, is an analytical technique that separates liquid samples under high pressures. An HPLC instrument consists of glass bottles for storing solvents called mobile phase reservoirs. HPLC-grade solvents are used to maintain high purity, and the dissolved gases are removed using a degasser, such as a vacuum pumping system or sparging with helium. The solvents are then pumped into the analytical column using a screw-driven syringe or reciprocating pumps.

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相关实验视频

Updated: Jun 29, 2026

Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification
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多列纳米流液体染色学与加速离线梯度生成,用于稳固和敏感的单细胞蛋白质组概况.

Xiaofeng Xie1,2, Thy Truong1,2, Siqi Huang1

  • 1Department of Chemistry and Biochemistry, Brigham Young University, Provo, Utah 84602, United States.

Analytical chemistry
|June 25, 2024
PubMed
概括

我们开发了一种用于高通量蛋白质组学的多列纳米LC系统,使单细胞的快速分析成为可能. 这种强大的系统提高了灵敏度,并减少了复杂分离的维护.

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科学领域:

  • 蛋白质组学是指蛋白质组学.
  • 分析化学 分析化学
  • 生物技术是生物技术.

背景情况:

  • 高灵敏度,强度,峰值容量和吞吐量对于自下而上的蛋白质组学至关重要,特别是对于分析像单细胞这样的小样本.
  • 当前的方法在应用于复杂的蛋白质组分析时,面临速度和维护方面的限制.

研究的目的:

  • 开发一种先进的多列纳米液态染色学 (nanoLC) 系统,具有离线梯度生成,用于增强蛋白质组学.
  • 为了提高单细胞蛋白质组分析的分离的效率,稳定性和吞吐量.

主要方法:

  • 设计了一个多列纳米LC系统,使用支持多个分析列的二进制进行线下梯度生成.
  • 为了减少维护和简化故障排除,实施了一个单一的陷列,与所有分析列接口.
  • 从陷柱进入样本循环的选择性离线化被用来保护分析柱免受污染物.

主要成果:

  • 该系统可在40nL/min流速下每20分钟进行一次样本分析,近100%的质谱利用率.
  • 实现了几个月的连续运行,没有更换柱子,大大提高了柱子的寿命和系统的稳定性.
  • 该系统成功地用于分析多发性骨髓瘤细胞系中单细胞的蛋白质组,该细胞系被用lenalidomide治疗.

结论:

  • 开发的多列纳米LC系统为自下而上的蛋白质组学提供了高度灵敏,坚固和高通量解决方案.
  • 这项技术显著提高了从有限的生物样本 (如单细胞) 中分析复杂蛋白质组的能力.
  • 该系统的设计有利于并行处理和延长运行稳定性,使其适合长期蛋白质组学研究.