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

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: 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...
Analyte Adsorption and Distribution01:09

Analyte Adsorption and Distribution

In certain chromatographic separations, solutes transfer between the mobile phase and the stationary phase via sorption, which typically refers to the process of adsorption. For many chromatographic systems, the sorption process often depends on the polarity of the compounds—an expression of the overall dipole moment within the molecule. During the separation process, there is competition between the solute and solvent for adsorption to the stationary phase. Highly polar compounds and solvents...
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

Temperature-Responsive Liquid Chromatography for One- and Two-Dimensional Separations in Compound-Specific Isotope Analysis.

Analytical chemistry·2025
Same author

The Hyphenation of High-Performance Liquid Chromatography with X-ray Fluorescence for Universal, Flow-Through, Elemental Analysis of Organobromines.

Analytical chemistry·2025
Same author

Combining per-aqueous and chiral reversed phase separation modes towards an enhanced comprehensive 2-dimensional liquid chromatographic based chiral screening platform.

Journal of chromatography. A·2025
Same author

Prediction of Retention Indices in LC-HRMS for Enhanced Structural Identification of Organic Micropollutants in Water: Selectivity-Based Filtration.

Analytical chemistry·2025
Same author

Maximizing sensitivity and selectivity in LC × LC-HRMS for pesticide analysis via exploitation of per-aqueous liquid chromatography.

Journal of chromatography. A·2024
Same author

2D-CEX-FcRn-MS to Study Structure/Function Relation of mAb Charge Variants.

Analytical chemistry·2024
Same journal

The ACS at 150: The History of Analytical Chemistry Publications and a Century of Progress.

Analytical chemistry·2026
Same journal

Machine Learning-Enabled Image Analysis of Complex Chemical Mixtures: Synthetic Urine Droplets as a Test System.

Analytical chemistry·2026
Same journal

H<sub>2</sub>O<sub>2</sub>/Viscosity Tandem-Locked Fluorescent Probes Based on an In Situ Fluorophore Synthesis Strategy for Colitis Imaging and Diagnosis.

Analytical chemistry·2026
Same journal

TopoStitcher: A Geometric-Topological Structure-Guided Stitching Framework for Single-Molecule Localization Microscopy.

Analytical chemistry·2026
Same journal

Noninvasive SERS Immunosensing of Tyrosinase for Melanoma Monitoring via Microneedle Sampling Integrated with Satellite-Structured Bifunctional Nanozymes.

Analytical chemistry·2026
Same journal

Label-Free Electrochemical CRISPR Platform Gated by Allosteric Transcription Factors for Ultrasensitive Small-Molecule Detection.

Analytical chemistry·2026
See all related articles

Related Experiment Video

Updated: Jun 16, 2026

Automated Hydrophobic Interaction Chromatography Column Selection for Use in Protein Purification
10:21

Automated Hydrophobic Interaction Chromatography Column Selection for Use in Protein Purification

Published on: September 21, 2011

Stationary-phase optimized selectivity liquid chromatography: development of a linear gradient prediction algorithm.

Maarten De Beer1, Fréderic Lynen, Kai Chen

  • 1Pfizer Analytical Research Centre, Ghent University, Krijgslaan 281-S4 Bis, B-9000 Ghent, Belgium.

Analytical Chemistry
|February 12, 2010
PubMed
Summary
This summary is machine-generated.

Stationary-phase optimized selectivity liquid chromatography (SOS-LC) now offers linear gradient optimization for complex mixtures. This advancement enhances reversed-phase LC (RP-LC) separations, improving analytical efficiency.

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

Liquid Chromatography Coupled to Refractive Index or Mass Spectrometric Detection for Metabolite Profiling in Lysate-based Cell-free Systems
14:42

Liquid Chromatography Coupled to Refractive Index or Mass Spectrometric Detection for Metabolite Profiling in Lysate-based Cell-free Systems

Published on: September 23, 2021

Related Experiment Videos

Last Updated: Jun 16, 2026

Automated Hydrophobic Interaction Chromatography Column Selection for Use in Protein Purification
10:21

Automated Hydrophobic Interaction Chromatography Column Selection for Use in Protein Purification

Published on: September 21, 2011

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

Curtain Flow Column: Optimization of Efficiency and Sensitivity

Published on: June 12, 2016

Liquid Chromatography Coupled to Refractive Index or Mass Spectrometric Detection for Metabolite Profiling in Lysate-based Cell-free Systems
14:42

Liquid Chromatography Coupled to Refractive Index or Mass Spectrometric Detection for Metabolite Profiling in Lysate-based Cell-free Systems

Published on: September 23, 2021

Area of Science:

  • Analytical Chemistry
  • Chromatography

Background:

  • Stationary-phase optimized selectivity liquid chromatography (SOS-LC) is a valuable tool for optimizing separations in reversed-phase LC (RP-LC) by using multisegment columns.
  • Current SOS-LC methods and algorithms are limited to isocratic analyses, and while step gradient SOS-LC exists, it's less effective for complex mixtures with wide hydrophobicity ranges.

Purpose of the Study:

  • To develop a linear gradient prediction algorithm for SOS-LC, enabling its application as a universal RP-LC optimization method.
  • To extend SOS-LC capabilities beyond isocratic and step gradient modes to include linear gradient runs.

Main Methods:

  • Development of a novel linear gradient prediction algorithm for SOS-LC.
  • Implementation of the algorithm to support isocratic, stepwise, and linear gradient run modes.
  • Experimental validation using a mixture of 13 steroids.

Main Results:

  • The developed algorithm successfully allows SOS-LC to operate in isocratic, stepwise, and linear gradient modes.
  • Baseline separation of a complex mixture of 13 steroids was predicted using linear gradient SOS-LC.
  • Experimental results confirmed the predicted baseline separation, demonstrating the algorithm's efficacy.

Conclusions:

  • A new linear gradient prediction algorithm significantly expands the utility of SOS-LC as a generic optimization method in RP-LC.
  • This advancement provides an elegant solution for analyzing complex mixtures with broad hydrophobicity ranges using SOS-LC.
  • The demonstrated baseline separation of steroids highlights the practical applicability of linear gradient SOS-LC.