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High-Performance Liquid Chromatography: Instrumentation

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: Introduction

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 Process

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...

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Mapping Dysfunctional Protein-Protein Interactions in Disease

Mapping Dysfunctional Protein-Protein Interactions in Disease

Published on: October 24, 2025

A practical guide to nano-LC troubleshooting.

Marek Noga¹, Filip Sucharski, Piotr Suder

¹Department of Neurobiochemistry, Faculty of Chemistry and Regional Laboratory, Jagiellonian University, Krakow, Poland.

Journal of Separation Science

|August 9, 2007

Summary

Troubleshooting capillary liquid chromatography (LC) systems is crucial for reliable results. This paper offers practical solutions for common issues like void volumes and injection problems in capillary LC.

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Area of Science:

Analytical Chemistry
Separation Science

Background:

Capillary LC offers powerful analytical capabilities.
Technical challenges can hinder capillary LC system operation and data reliability.

Purpose of the Study:

To provide practical guidance on diagnosing and resolving common capillary LC issues.
To present solutions for problems including void/dead volumes, leakages, and sample injection.

Main Methods:

Review and summarization of common capillary LC system problems.
Discussion of troubleshooting strategies and solutions.
Inclusion of exemplary nano-LC separation examples.

Main Results:

Identification of key troubleshooting areas in capillary LC systems.
Practical advice and solutions for system diagnosis and problem resolution.
Demonstration of typical issues through nano-LC examples.

Conclusions:

Effective troubleshooting is essential for maintaining capillary LC system performance.
Addressing issues like void volumes and injection problems ensures reliable analytical outcomes.
This guide aids scientists in overcoming daily operational challenges in capillary LC.