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: 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...
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.
Supercritical Fluid Chromatography01:18

Supercritical Fluid Chromatography

Supercritical fluid chromatography (SFC) provides a beneficial substitute for gas chromatography (GC) and liquid chromatography (LC) for certain samples because it merges the top attributes of both techniques. SFC allows the separation and analysis of compounds that GC or LC does not easily manage. These compounds are traditionally nonvolatile or thermally unstable, making GC unsuitable and lacking functional groups required for HPLC analysis.
SFC utilizes a supercritical fluid mobile phase,...
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...
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...

You might also read

Related Articles

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

Sort by
Same author

Development of analytical functions with high-performance liquid phase separation systems on the basis of the moment theory.

Analytical sciences : the international journal of the Japan Society for Analytical Chemistry·2025
Same author

Moment analysis method for the determination of permeation kinetics of coumarin at lipid bilayers of liposomes by using capillary electrophoresis.

Electrophoresis·2024
Same author

Moment analysis method for determination of rate constants of solute permeation across interface of spherical molecular aggregates by means of high-performance liquid chromatography.

Journal of chromatography. A·2024
Same author

Moment Analysis Method for Measurement of Reaction Equilibrium and Rate Constants by Using High-Performance Liquid Chromatography.

Analytical chemistry·2024
Same author

A study on attempt for determination of permeation kinetics of coumarin at lipid bilayer of liposomes by using capillary electrophoresis with moment analysis theory.

Journal of chromatography. A·2022
Same author

Moment theory of affinity capillary electrophoresis for analysis of reaction kinetics of intermolecular interactions.

Journal of chromatography. A·2022

Related Experiment Video

Updated: Jun 14, 2026

Improving High Viscosity Extrusion of Microcrystals for Time-resolved Serial Femtosecond Crystallography at X-ray Lasers
07:26

Improving High Viscosity Extrusion of Microcrystals for Time-resolved Serial Femtosecond Crystallography at X-ray Lasers

Published on: February 28, 2019

Kinetic study on external mass transfer in high performance liquid chromatography system.

Kanji Miyabe1, Yuuki Kawaguchi, Georges Guiochon

  • 1Graduate School of Science and Engineering for Research, University of Toyama, 3190, Gofuku, Toyama 930-8555, Japan.

Journal of Chromatography. A
|March 30, 2010
PubMed
Summary

External mass transfer coefficients were determined for C(18)-silica particles using pulse response and peak-parking methods. Correlations for Sherwood, Reynolds, and Schmidt numbers were validated for HPLC systems, confirming the accuracy of literature equations for small particles.

More Related Videos

Novel Techniques for Observing Structural Dynamics of Photoresponsive Liquid Crystals
10:35

Novel Techniques for Observing Structural Dynamics of Photoresponsive Liquid Crystals

Published on: May 29, 2018

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 14, 2026

Improving High Viscosity Extrusion of Microcrystals for Time-resolved Serial Femtosecond Crystallography at X-ray Lasers
07:26

Improving High Viscosity Extrusion of Microcrystals for Time-resolved Serial Femtosecond Crystallography at X-ray Lasers

Published on: February 28, 2019

Novel Techniques for Observing Structural Dynamics of Photoresponsive Liquid Crystals
10:35

Novel Techniques for Observing Structural Dynamics of Photoresponsive Liquid Crystals

Published on: May 29, 2018

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:

  • Chromatography
  • Chemical Engineering
  • Physical Chemistry

Background:

  • Accurate mass transfer coefficients are crucial for optimizing chromatographic separations.
  • Understanding external mass transfer is key to predicting band broadening in HPLC.
  • Previous studies have established correlations for mass transfer in packed beds, but validation for micro-scale particles is ongoing.

Purpose of the Study:

  • To measure external mass transfer coefficients (k(f)) for C(18)-silica particles in HPLC.
  • To investigate the relationship between Sherwood (Sh), Reynolds (Re), and Schmidt (Sc) numbers under varying flow rates.
  • To validate existing literature correlations for mass transfer in packed beds with micro-scale particles.

Main Methods:

  • Utilized pulse response and peak-parking chromatography techniques.
  • Employed C(18)-silica spherical particles (50.6 µm diameter) with a methanol/water (70/30, v/v) mobile phase.
  • Analyzed elution peak profiles and moments to determine k(f), intraparticle diffusion, and surface diffusion coefficients.

Main Results:

  • Measured Sh, Re, and Sc numbers within ranges of 0.004–0.05, 1.8x10^3–2.7x10^3, respectively.
  • Found Sh to be proportional to Re^α and Sc^β, with exponents α (0.26–0.41) and β (0.31–0.36) consistent with literature.
  • Demonstrated that conventional literature correlations accurately estimate k(f) for micro-scale HPLC particles with a ~15% error using the Wilson-Geankoplis equation.

Conclusions:

  • External mass transfer coefficients in HPLC can be reliably determined using pulse response and peak-parking methods.
  • Established correlations between Sh, Re, and Sc numbers are applicable to micro-scale C(18)-silica particles.
  • Conventional correlations provide accurate estimations of k(f) in HPLC, even for sub-50 micrometer particles, supporting their use in system design and optimization.