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Related Concept Videos

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: 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.
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...
Chromatographic Methods: Terminology01:18

Chromatographic Methods: Terminology

Chromatography is an analytical technique widely used in fields such as chemistry, biology, environmental science, and pharmaceuticals to separate the components of a mixture and identify substances between them. The process of chromatography is based on the interactions between two distinct phases: the stationary phase and the mobile phase. The stationary phase is fixed in place by a supporting material, while the mobile phase moves over it, carrying the solutes. As the mobile phase travels,...
Supercritical Fluid Chromatography01:18

Supercritical Fluid Chromatography

Supercritical fluid chromatography (SFC) provides a beneficial substitute for gas chromatography (GC) and liquid chromatography (LC) for certain samples because it merges the top attributes of both techniques. SFC allows the separation and analysis of compounds that GC or LC does not easily manage. These compounds are traditionally nonvolatile or thermally unstable, making GC unsuitable and lacking functional groups required for HPLC analysis.
SFC utilizes a supercritical fluid mobile phase,...
Chromatographic Methods: Classification01:12

Chromatographic Methods: Classification

Chromatographic techniques are classified in three ways: the classification is based on the physical state of the stationary and mobile phases, how the mobile phase and the stationary phase contact each other, or through the chemical or physical processes that isolate the components of the sample. Typically, the mobile phase is either a liquid or gas, while the stationary phase is either a solid or a liquid layer applied to a solid surface.
Chromatographic techniques are typically named by...

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Automated Hydrophobic Interaction Chromatography Column Selection for Use in Protein Purification
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Published on: September 21, 2011

A Protocol for Characterizing Comprehensive Two-Dimensional Liquid Chromatography Systems.

Megane K Aebischer1,2, Marie Pardon3,4, Clémence Gadot5

  • 1School of Pharmaceutical Sciences, University of Geneva, Geneva, Switzerland.

Journal of Separation Science
|May 11, 2026
PubMed
Summary
This summary is machine-generated.

This study presents a practical protocol for characterizing comprehensive two-dimensional liquid chromatography (LC × LC) systems. System characterization is crucial for optimizing LC × LC methods and improving separation performance for complex samples.

Keywords:
2D‐LCLC × LCbidimensional liquid chromatographycharacterizationsystem dispersion

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Area of Science:

  • Analytical Chemistry
  • Chromatography

Background:

  • Comprehensive two-dimensional liquid chromatography (LC × LC) enhances separation performance for complex samples compared to 1D-LC.
  • Method development in LC × LC is challenging due to interconnected thermodynamic and kinetic parameters.

Purpose of the Study:

  • To propose a practical protocol for comprehensive 2D-LC system characterization.
  • To provide a guide for optimizing LC × LC instruments and methods.

Main Methods:

  • Developed a protocol for characterizing 2D-LC instrumentation.
  • Utilized the 2D-LC Smart Calculator software for calculations.
  • Validated the protocol through inter-laboratory testing and in silico performance evaluation.

Main Results:

  • The proposed protocol facilitates system-specific characterization for reliable LC × LC implementation.
  • Instrumental features significantly impact LC × LC performance, as shown by in silico evaluations.
  • Inter-laboratory testing confirmed the protocol's usability.

Conclusions:

  • System-specific characterization is essential for successful LC × LC method development and optimization.
  • The protocol and associated software aid users in understanding and optimizing their 2D-LC systems.
  • Practical recommendations are provided for instrument adaptation and performance enhancement.