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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.
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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.

Principles Of Column Chromatography

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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...

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Related Experiment Video

Updated: May 16, 2026

Fabrication of the Thermoplastic Microfluidic Channels

Fabrication of the Thermoplastic Microfluidic Channels

Published on: February 3, 2008

Fabrication of high-quality microfluidic solid-phase chromatography columns.

Jens Huft¹, Charles A Haynes, Carl L Hansen

¹Centre for High-Throughput Biology, University of British Columbia, Vancouver, British Columbia, Canada.

Analytical Chemistry

|December 14, 2012

Summary

A new low-pressure bead packing method enables high-performance liquid chromatography (HPLC) columns in microfluidic devices. This technique ensures robust integration and exceptional efficiency for analytical separations in chemistry and biology.

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Simple In-House Ultra-High Performance Capillary Column Manufacturing with the FlashPack Approach

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Published on: December 4, 2021

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Last Updated: May 16, 2026

Fabrication of the Thermoplastic Microfluidic Channels

Fabrication of the Thermoplastic Microfluidic Channels

Published on: February 3, 2008

Simple In-House Ultra-High Performance Capillary Column Manufacturing with the FlashPack Approach

Simple In-House Ultra-High Performance Capillary Column Manufacturing with the FlashPack Approach

Published on: December 4, 2021

Area of Science:

Microfluidics
Analytical Chemistry
Materials Science

Background:

Integrating high-performance chromatography columns into microfluidic devices presents challenges.
Existing methods may lack robustness or efficiency for advanced analytical applications.

Purpose of the Study:

To develop a novel low-pressure bead packing technique for robust integration of high-performance chromatography columns in poly(dimethylsiloxane) microfluidic devices.
To demonstrate the efficiency and reproducibility of the packed microfluidic columns.

Main Methods:

Utilized a low-pressure bead packing technique.
Employed multilayer soft lithography (MSL) for microfluidic device fabrication.
Incorporated a novel column geometry with micrometer-sized bypass channels.
Performed pulse tests to evaluate column performance.

Main Results:

Achieved rapid packing of multiple high-quality bead bed columns in parallel with near-perfect yield.
Demonstrated exceptional reproducibility and efficiency in microfluidic columns.
Measured plate counts of 1,650,000/m ± 7% and a reduced plate height of h = 0.12 ± 7%.

Conclusions:

The developed low-pressure bead packing technique enables robust integration of high-performance chromatography columns in microfluidic devices.
The novel column geometry and packing method result in highly efficient and reproducible microfluidic chromatography.
This advancement offers new possibilities for integrated sample processing and separation in biological and chemical analyses.