Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

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...
Optimizing Chromatographic Separations01:15

Optimizing Chromatographic Separations

Optimizing chromatographic separations is crucial for obtaining clean separations in a minimum amount of time. Optimization is required for several factors, including kinetic effects related to band broadening, plate height, capacity factor, and separation factor.
Band broadening refers to spreading solute bands as they travel through the column. This broadening can impact resolution. Plate height (H) represents the length required for one theoretical plate. A lower plate height corresponds to...
High-Performance Liquid Chromatography: Types of Detectors01:15

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...
Principles Of Column Chromatography01:13

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...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Uncertainty-driven model-based search methods for method development in liquid chromatography.

Journal of chromatography. A·2026
Same author

Automated micropillar array design with Bayesian optimization and computational fluid dynamics.

Journal of chromatography. A·2026
Same author

Computational study of the mass transfer effects in metal-organic framework columns for liquid chromatography.

Journal of chromatography. A·2026
Same author

Revisiting the definition and measurement of total accessible and hold-up volumes in liquid chromatography: Current understanding and advances.

Journal of chromatography. A·2026
Same author

SERS Substrate Fabrication via Rapid Triboelectrification-Driven Self-Assembly of Close-Packed Colloidal Monolayers.

Small methods·2026
Same author

Kinetic plot method for the optimal column and instrument design for high speed chiral SFC applications.

Journal of chromatography. A·2026
Same journal

Chromatographic Purification of Complex Natural Products as a Decision Problem: Retention Prediction, Adaptive Optimization, and Experimental Feedback.

Journal of separation science·2026
Same journal

A High-Throughput Analytical Approach Using Polyaniline Doped With Oxalic Acid in Thin Film Solid-Phase Microextraction for the Determination of Personal Care Products in Recreational Waters.

Journal of separation science·2026
Same journal

Simultaneous Determination of Dechlorane-Related Compounds in Fish Muscle by Microwave-Assisted Extraction Combined With Enhanced Matrix Removal for Lipids Cleanup and Gas Chromatography-Tandem Mass Spectrometry.

Journal of separation science·2026
Same journal

Covalent Zwitterionic Peptide-Based Antifouling Coating of the Fused Silica Capillary Applied for CE Separation of Proteins.

Journal of separation science·2026
Same journal

Pharmacokinetic Study of Five Lead Components of Psoraleae Fructus in Human Serum by UHPLC-Q-TOF-MS and UHPLC-QqQ-MS/MS after Oral Administration of Qing'e Pills.

Journal of separation science·2026
Same journal

Passive Blood-Plasma Separation via Constriction-Expansion Geometry in Untreated Paper Microfluidic Devices.

Journal of separation science·2026
See all related articles

Related Experiment Video

Updated: Jun 3, 2026

Post Column Derivatization Using Reaction Flow High Performance Liquid Chromatography Columns
06:25

Post Column Derivatization Using Reaction Flow High Performance Liquid Chromatography Columns

Published on: April 26, 2016

Kinetic performance optimisation for liquid chromatography: principles and practice.

Tim J Causon1, Ken Broeckhoven, Emily F Hilder

  • 1Australian Centre for Research on Separation Science, School of Chemistry, University of Tasmania, Hobart, Australia.

Journal of Separation Science
|March 12, 2011
PubMed
Summary
This summary is machine-generated.

This tutorial introduces kinetic plots for optimizing High-Performance Liquid Chromatography (HPLC) performance. Learn to select columns and conditions for maximum efficiency in minimal time.

More Related Videos

Untargeted Metabolomics from Biological Sources Using Ultraperformance Liquid Chromatography-High Resolution Mass Spectrometry (UPLC-HRMS)
11:00

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

Published on: May 20, 2013

Curtain Flow Column: Optimization of Efficiency and Sensitivity
06:44

Curtain Flow Column: Optimization of Efficiency and Sensitivity

Published on: June 12, 2016

Related Experiment Videos

Last Updated: Jun 3, 2026

Post Column Derivatization Using Reaction Flow High Performance Liquid Chromatography Columns
06:25

Post Column Derivatization Using Reaction Flow High Performance Liquid Chromatography Columns

Published on: April 26, 2016

Untargeted Metabolomics from Biological Sources Using Ultraperformance Liquid Chromatography-High Resolution Mass Spectrometry (UPLC-HRMS)
11:00

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

Published on: May 20, 2013

Curtain Flow Column: Optimization of Efficiency and Sensitivity
06:44

Curtain Flow Column: Optimization of Efficiency and Sensitivity

Published on: June 12, 2016

Area of Science:

  • Analytical Chemistry
  • Chromatography

Background:

  • High-Performance Liquid Chromatography (HPLC) is crucial for chemical analysis.
  • Optimizing HPLC performance is key for efficient and timely separations.
  • Kinetic plots offer a graphical method for performance characterization.

Purpose of the Study:

  • To provide a tutorial on preparing and using kinetic plots in HPLC.
  • To demonstrate how kinetic plots aid in selecting optimal columns and LC conditions.
  • To discuss instrument parameters affecting HPLC performance.

Main Methods:

  • Utilizing kinetic plots for performance characterization in isocratic and gradient LC.
  • Evaluating column selection based on particle size and length.
  • Assessing LC conditions like operating pressure and temperature.
  • Analyzing instrument factors such as extra-column effects and thermal conditions.

Main Results:

  • Kinetic plots enable the selection of optimal columns and LC conditions for desired plate number or peak capacity.
  • Analysis time can be minimized by strategic selection of parameters.
  • Instrumental aspects like system volume and oven type significantly impact performance.
  • Porous-shell and monolithic columns exhibit distinct performance characteristics.

Conclusions:

  • Kinetic plots are a valuable tool for optimizing HPLC method development.
  • Informed selection of columns and instrument parameters leads to enhanced analytical efficiency.
  • Understanding instrumental limitations is vital for achieving optimal separation performance.