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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.
In HPLC, two phases play a critical role in the separation process:

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...

High-Performance Liquid Chromatography: Types of Detectors

High-Performance Liquid Chromatography: Types of Detectors

The role of the detectors in High-Performance Liquid Chromatography (HPLC) is to analyze the solutes as they exit from the chromatographic column. The detector recognizes the solute's property and generates corresponding electrical signals, which are converted into a readable graph of the detector's response versus elution time called a chromatogram at the computer. There are several types of HPLC detectors, each with its own advantages and limitations, depending on the analyte properties and...

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Related Experiment Video

Updated: May 22, 2026

Untargeted Metabolomics from Biological Sources Using Ultraperformance Liquid Chromatography-High Resolution Mass Spectrometry (UPLC-HRMS)

Untargeted Metabolomics from Biological Sources Using Ultraperformance Liquid Chromatography-High Resolution Mass Spectrometry (UPLC-HRMS)

Published on: May 20, 2013

UPLC: a preeminent technique in pharmaceutical analysis.

Ashok Kumar¹, Gautam Saini, Anroop Nair

¹M. M. College of Pharmacy, M. M. University, Mullana, Ambala, Haryana-133207, India. ashokmmcp@rediffmail.com

Acta Poloniae Pharmaceutica

|May 19, 2012

Summary

Ultra-high performance liquid chromatography (UPLC) offers significant advancements over traditional HPLC, enhancing speed, resolution, and sensitivity in drug discovery and development. This review explores UPLC

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Large Scale Non-targeted Metabolomic Profiling of Serum by Ultra Performance Liquid Chromatography-Mass Spectrometry (UPLC-MS)

Large Scale Non-targeted Metabolomic Profiling of Serum by Ultra Performance Liquid Chromatography-Mass Spectrometry (UPLC-MS)

Published on: March 14, 2013

Related Experiment Videos

Last Updated: May 22, 2026

Untargeted Metabolomics from Biological Sources Using Ultraperformance Liquid Chromatography-High Resolution Mass Spectrometry (UPLC-HRMS)

Untargeted Metabolomics from Biological Sources Using Ultraperformance Liquid Chromatography-High Resolution Mass Spectrometry (UPLC-HRMS)

Published on: May 20, 2013

Large Scale Non-targeted Metabolomic Profiling of Serum by Ultra Performance Liquid Chromatography-Mass Spectrometry (UPLC-MS)

Large Scale Non-targeted Metabolomic Profiling of Serum by Ultra Performance Liquid Chromatography-Mass Spectrometry (UPLC-MS)

Published on: March 14, 2013

Area of Science:

Analytical Chemistry
Pharmaceutical Science

Background:

Pharmaceutical companies require advanced analytical tools for efficient drug discovery and development.
Technological progress in chromatography has led to enhanced performance in analytical laboratories.

Purpose of the Study:

To review the fundamental principles and instrumentation of Ultra-High Performance Liquid Chromatography (UPLC).
To highlight the advantages of UPLC compared to High-Performance Liquid Chromatography (HPLC).
To emphasize the diverse pharmaceutical applications of UPLC.

Main Methods:

Review of technological advancements in chromatographic techniques.
Focus on particle chemistry, detector design, and system optimization.
Comparison of UPLC operational parameters (particle size, pressure, linear velocity) with HPLC.

Main Results:

UPLC utilizes sub-2 µm particles and operates at higher pressures, enabling greater linear velocities.
UPLC demonstrates dramatic improvements in analysis speed, resolution, and sensitivity over HPLC.
UPLC represents a significant focal point in liquid chromatography studies.

Conclusions:

UPLC offers superior performance characteristics compared to HPLC for pharmaceutical analysis.
The technique's enhanced speed, resolution, and sensitivity make it crucial for modern drug development.
UPLC is a pivotal advancement in liquid chromatographic studies with broad pharmaceutical applications.