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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: 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...

Principles Of Column Chromatography

Principles Of Column Chromatography

The chromatography technique was first invented in 1901 by Michael S. Tswett, a Russian botanist, to separate plant pigments using organic solvents. Further, in 1941, Archer John Porter Martin and R. L. M. Synge modified the technique by packing silica gel into a column. A mixture of amino acids was then separated on the packed column using chloroform and water mixture as the mobile phase. This was the first report on column chromatography. At present, column chromatography is a widely used...

Ion-Exchange Chromatography

Ion-Exchange Chromatography

Ion-exchange chromatography, or IEC, is a technique for separating ions based on their affinity for the stationary phase. The stationary phase is a cross-linked polymer resin with covalently attached ionic functional groups. The functional groups can be either positively charged (cation exchangers) or negatively charged (anion exchangers). A cation exchanger consists of a polymeric anion and active cations, while an anion exchanger is a polymeric cation with active anions. The choice of...

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:

Types Of Column Chromatography

Types Of Column Chromatography

The stability and compatibility of column material with samples are crucial for efficient purification in chromatographic techniques. Various operating parameters such as pH, temperature, or solvent affect the packing of the column material, thereby determining the purification efficiency. The choice of column material also plays an essential role in deciding the operating parameters and can be modified based on the proteins that need to be purified.
Gel Filtration Chromatography
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Related Experiment Video

Updated: Jul 12, 2026

Automated Hydrophobic Interaction Chromatography Column Selection for Use in Protein Purification

Automated Hydrophobic Interaction Chromatography Column Selection for Use in Protein Purification

Published on: September 21, 2011

Development of a Hydraulic-Drive Automatic Column Chromatography System for High Precision Isotope Analysis.

Wenkai Zhang¹, Qilong Li¹, Yingran Liang¹

¹State Key Laboratory of Geomicrobiology and Environmental Changes, China University of Geosciences (Wuhan), Wuhan 430074, China.

Analytical Chemistry

|July 10, 2026

Summary

A novel hydraulic chromatography system offers metal-free, high-throughput isotope analysis. This automated, cost-effective platform ensures accurate separations, minimizing contamination for precise scientific measurements.

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Post Column Derivatization Using Reaction Flow High Performance Liquid Chromatography Columns

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Published on: April 26, 2016

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Last Updated: Jul 12, 2026

Automated Hydrophobic Interaction Chromatography Column Selection for Use in Protein Purification

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Published on: September 21, 2011

Post Column Derivatization Using Reaction Flow High Performance Liquid Chromatography Columns

Post Column Derivatization Using Reaction Flow High Performance Liquid Chromatography Columns

Published on: April 26, 2016

Area of Science:

Analytical Chemistry
Geochemistry
Environmental Science

Background:

Conventional chromatography systems risk metal contamination and corrosion from acidic solutions.
High-precision isotope analysis requires meticulous sample preparation to avoid errors.

Purpose of the Study:

To develop a high-throughput, automated, and metal-free chromatography system for precise isotope analysis.
To overcome limitations of conventional systems in handling corrosive materials.

Main Methods:

A hydraulic remote-drive mechanism controls eluent delivery, isolating metal components from corrosive environments.
Multichannel parallel elution (8 channels) with pressure sensing and mass flow feedback control.
Construction from low-cost polymers for scalability and affordability.

Main Results:

Achieved high accuracy (<3% error) and interchannel consistency (<0.5% variation) in separations.
Demonstrated high recoveries for Cd (97.5 ± 1.6%) and Cu (98.3 ± 1.4%) isotope separation.
Obtained isotopic ratios from geological reference materials consistent with published values, with low procedural blanks.

Conclusions:

The developed system is a scalable, cost-effective, and metal-free solution for automated sample preparation in isotope analysis.
It effectively mitigates contamination and corrosion issues, enhancing reliability for geological and environmental studies.
The platform supports high-throughput separations crucial for advancing high-precision isotopic measurements.