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:
Ion-Exchange Chromatography01:09

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: 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: 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.
Silica Gel Column Chromatography: Overview01:10

Silica Gel Column Chromatography: Overview

Silica gel column chromatography is a technique for separating compounds using a column packed with silica gel as the stationary phase. This method relies on differences in the polarity of compounds. Based on their polarities, compounds move between the stationary phase (silica gel) and the mobile phase (the solvent), forming discrete bands in the column.
Polar components tend to bind strongly to the silica gel, causing them to move slowly through the column. In contrast, nonpolar compounds...
Gas Chromatography: Types of Columns and Stationary Phases01:17

Gas Chromatography: Types of Columns and Stationary Phases

Gas chromatography (GC) relies on stationary phases to separate and analyze components in a sample. There are two main types of stationary phases: liquid and solid. Liquid stationary phases are non-volatile, thermally stable, and chemically inert liquids coated onto the column. Solid stationary phases are particles of adsorbent material, such as silica gel or molecular sieves.
For an analyte to remain on the column for a sufficient amount of time, it must exhibit some level of compatibility (or...

You might also read

Related Articles

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

Sort by
Same author

Patient-Specific MRI-Based Right Ventricle Models Using Different Zero-Load Diastole and Systole Geometries for Better Cardiac Stress and Strain Calculations and Pulmonary Valve Replacement Surgical Outcome Predictions.

PloS one·2016
Same author

Crossover phenomena of percolation transition in evolution networks with hybrid attachment.

Chaos (Woodbury, N.Y.)·2016
Same author

Relation Between C-X-C Motif Chemokine Receptor 4 Levels and the Presence and Extent of Angiographic Coronary Collaterals in Patients With Chronic Total Coronary Occlusion.

The American journal of cardiology·2016
Same author

Quantify patient-specific coronary material property and its impact on stress/strain calculations using in vivo IVUS data and 3D FSI models: a pilot study.

Biomechanics and modeling in mechanobiology·2016
Same author

Analysis of Genomewide DNA Methylation Reveals Differences in DNA Methylation Levels between Dormant and Naturally as well as Artificially Potentiated Pedicle Periosteum of Sika Deer (Cervus nippon).

Journal of experimental zoology. Part B, Molecular and developmental evolution·2016
Same author

Value of corneal epithelial and Bowman's layer vertical thickness profiles generated by UHR-OCT for sub-clinical keratoconus diagnosis.

Scientific reports·2016

Related Experiment Video

Updated: Jun 28, 2026

Simple In-House Ultra-High Performance Capillary Column Manufacturing with the FlashPack Approach
13:36

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

Published on: December 4, 2021

High-efficiency liquid chromatographic separation utilizing long monolithic silica capillary columns.

Kosuke Miyamoto1, Takeshi Hara, Hiroshi Kobayashi

  • 1Department of Biomolecular Engineering, Kyoto Institute of Technology, Matsugasaki, Sakyo-ku, Kyoto 606-8585, Japan.

Analytical Chemistry
|October 25, 2008
PubMed
Summary

Researchers developed long monolithic silica-C18 capillary columns for high-efficiency separations. These columns offer over 500,000 theoretical plates, enabling precise analysis of complex samples in reversed-phase high-performance liquid chromatography (HPLC).

More Related Videos

Improved Polymerase Chain Reaction-restriction Fragment Length Polymorphism Genotyping of Toxic Pufferfish by Liquid Chromatography/Mass Spectrometry
09:34

Improved Polymerase Chain Reaction-restriction Fragment Length Polymorphism Genotyping of Toxic Pufferfish by Liquid Chromatography/Mass Spectrometry

Published on: September 20, 2016

Related Experiment Videos

Last Updated: Jun 28, 2026

Simple In-House Ultra-High Performance Capillary Column Manufacturing with the FlashPack Approach
13:36

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

Published on: December 4, 2021

Improved Polymerase Chain Reaction-restriction Fragment Length Polymorphism Genotyping of Toxic Pufferfish by Liquid Chromatography/Mass Spectrometry
09:34

Improved Polymerase Chain Reaction-restriction Fragment Length Polymorphism Genotyping of Toxic Pufferfish by Liquid Chromatography/Mass Spectrometry

Published on: September 20, 2016

Area of Science:

  • Analytical Chemistry
  • Chromatography
  • Materials Science

Background:

  • High-performance liquid chromatography (HPLC) is crucial for chemical analysis.
  • Developing columns with enhanced efficiency and capacity is an ongoing challenge.
  • Monolithic silica capillary columns offer unique chromatographic properties.

Purpose of the Study:

  • To prepare and evaluate long monolithic silica-C18 capillary columns.
  • To assess the chromatographic efficiency and performance of these columns.
  • To demonstrate their utility in separating complex mixtures and biomolecules.

Main Methods:

  • Preparation of 100 microm internal diameter monolithic silica-C18 capillary columns.
  • Reversed-phase HPLC analysis under high pressure (up to 47 MPa).
  • Evaluation of column efficiency using theoretical plates and plate height.
  • Separation of aromatic hydrocarbons, benzene derivatives, and protein digests.

Main Results:

  • Generated 100,000–500,000 theoretical plates with single columns (90–440 cm).
  • Achieved a minimum plate height of 8.5 ± 0.5 microm and permeability of (1.45 ± 0.09) x 10(-13) m(2).
  • Connected columns achieved over 1,000,000 theoretical plates for aromatic hydrocarbons.
  • Maintained efficiency for peptide analysis with up to 0.5–2 ng injection.

Conclusions:

  • Long monolithic silica-C18 capillary columns provide exceptionally high chromatographic efficiency.
  • These columns demonstrate practical utility for high-resolution separations of diverse analytes.
  • The developed columns are suitable for demanding analytical applications, including proteomics.