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

Principles Of Column Chromatography01:13

Principles Of Column Chromatography

9.6K
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...
9.6K
High-Performance Liquid Chromatography: Introduction01:11

High-Performance Liquid Chromatography: Introduction

4.0K
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:
4.0K
High-Performance Liquid Chromatography: Elution Process01:05

High-Performance Liquid Chromatography: Elution Process

1.9K
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...
1.9K
High-Performance Liquid Chromatography: Instrumentation00:57

High-Performance Liquid Chromatography: Instrumentation

3.5K
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.
3.5K
Ion-Exchange Chromatography01:09

Ion-Exchange Chromatography

2.7K
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...
2.7K
Types Of Column Chromatography01:29

Types Of Column Chromatography

15.0K
The stability and compatibility of column material with samples are crucial for efficient purification in chromatographic techniques. Various operating parameters such as pH, temperature, or solvent affect the packing of the column material, thereby determining the purification efficiency. The choice of column material also plays an essential role in deciding the operating parameters and can be modified based on the proteins that need to be purified.
Gel Filtration Chromatography
When the...
15.0K

You might also read

Related Articles

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

Sort by
Same author

Accuracy of menu calorie labelling in the England out-of-home food sector during 2024: assessment of a national food policy.

The British journal of nutrition·2025
Same author

An Ecological Momentary Assessment Study of Fluctuations in Inhibitory Control and Its Predictive Validity of Alcohol Use.

Substance use & misuse·2025
Same author

Comparing the impact of feedback on excess calories and physical activity calorie equivalent (PACE) information on consumer behaviour in an online hypothetical restaurant setting.

Appetite·2025
Same author

Elucidating tissue and subcellular specificity of the entire SUMO network reveals how stress responses are fine-tuned in a eukaryote.

Science advances·2025
Same author

Social influence increases the value and consumption of alcohol in the laboratory.

Alcohol, clinical & experimental research·2025
Same author

Cities Are Concentrators of Complex, MultiSectoral Interactions Within the Human-Earth System.

Earth's future·2025
Same journal

Reliability of A Vibration-Based Elastography Protocol For Assessing Achilles Tendon Stiffness Across Multiple Joint Angles In Elite Athletes.

Journal of visualized experiments : JoVE·2026
Same journal

Associations of Inflammatory and Coagulation Biomarkers with Kidney Injury Across Chronic and Acute Clinical Settings.

Journal of visualized experiments : JoVE·2026
Same journal

Intelligent Recommender Systems for Chinese Super League Fan Consumption Behavior Prediction.

Journal of visualized experiments : JoVE·2026
Same journal

A Battery of Quantitative Binocular Vision Tests for Adults: Testing Protocols.

Journal of visualized experiments : JoVE·2026
Same journal

Efficacy Analysis of Paiteling in Treating Persistent High-Risk Human Papillomavirus after Cervical Cancer Surgery.

Journal of visualized experiments : JoVE·2026
Same journal

Clinical Efficacy of Tissue-Bone Homeostasis Manipulation on Soft Tissue Balance and Function in Knee Osteoarthritis.

Journal of visualized experiments : JoVE·2026
See all related articles

Related Experiment Video

Updated: Mar 21, 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

16.0K

Post Column Derivatization Using Reaction Flow High Performance Liquid Chromatography Columns.

Andrew Jones1, Sercan Pravadali-Cekic1, Stanley Hua1

  • 1School of Science and Health, University of Western Sydney.

Journal of Visualized Experiments : Jove
|May 12, 2016
PubMed
Summary
This summary is machine-generated.

A new protocol for reaction flow post-column derivatization (RF-PCD) in high-performance liquid chromatography minimizes band broadening. This technique enhances separation efficiency and detection sensitivity compared to traditional methods.

More Related Videos

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

Curtain Flow Column: Optimization of Efficiency and Sensitivity

Published on: June 12, 2016

7.0K
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

4.7K

Related Experiment Videos

Last Updated: Mar 21, 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

16.0K
Curtain Flow Column: Optimization of Efficiency and Sensitivity
06:44

Curtain Flow Column: Optimization of Efficiency and Sensitivity

Published on: June 12, 2016

7.0K
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

4.7K

Area of Science:

  • Analytical Chemistry
  • Chromatography

Background:

  • Traditional post-column derivatization (PCD) in HPLC faces challenges with large reaction coils, leading to band broadening and reduced sensitivity.
  • Adapting PCD to modern HPLC systems requires minimizing post-column dead volume to maintain separation efficiency.

Purpose of the Study:

  • To present a protocol for reaction flow post-column derivatization (RF-PCD) using specialized HPLC columns.
  • To address the limitations of conventional PCD by reducing dead volume and improving analytical performance.

Main Methods:

  • Utilized reaction flow HPLC columns with integrated mixing frits in the end fitting.
  • Pumped derivatization reagents into outer ports of the reaction flow column, mixing against the mobile phase flow.
  • Minimized or eliminated the need for large reaction coils.

Main Results:

  • RF-PCD demonstrated improved separation efficiency and signal-to-noise ratio compared to conventional PCD.
  • The technique allows for monitoring of underivatized effluent from a central port.
  • RF-PCD effectively minimized reaction coil volumes.

Conclusions:

  • RF-PCD offers a superior alternative to conventional PCD for HPLC methods.
  • The technique shows potential for broad applicability across various post-column derivatization reactions.
  • RF-PCD enhances analytical performance by reducing band broadening and improving sensitivity.