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

Ion-Exchange Chromatography01:09

Ion-Exchange Chromatography

826
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...
826
Chromatographic Methods: Terminology01:18

Chromatographic Methods: Terminology

2.6K
Chromatography is an analytical technique widely used in fields such as chemistry, biology, environmental science, and pharmaceuticals to separate the components of a mixture and identify substances between them. The process of chromatography is based on the interactions between two distinct phases: the stationary phase and the mobile phase. The stationary phase is fixed in place by a supporting material, while the mobile phase moves over it, carrying the solutes. As the mobile phase travels,...
2.6K
Principles Of Column Chromatography01:13

Principles Of Column Chromatography

7.3K
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...
7.3K
High-Performance Liquid Chromatography: Introduction01:11

High-Performance Liquid Chromatography: Introduction

2.4K
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:
2.4K
Peptide Identification Using Tandem Mass Spectrometry01:33

Peptide Identification Using Tandem Mass Spectrometry

7.0K
Tandem mass spectrometry, also known as MS/MS or MS2, is an analytical technique that employs two mass analyzers. Essentially it is a series of mass spectrometers that helps isolate a particular biomolecule and then helps study its chemical properties.
This technique helps gather information regarding the protein from which the peptide was obtained and to study the peptides’ amino acid sequence. Identifying peptides from a complex mixture is an important component of the growing field of...
7.0K
Affinity Chromatography01:03

Affinity Chromatography

1.4K
Affinity chromatography is a powerful technique extensively utilized for separating and purifying specific biomolecules from complex mixtures. It capitalizes on the highly selective binding between an analyte and its counterpart, such as antibody-antigen interactions. The counterpart is immobilized on the stationary phase, forming an affinity column. The stationary phase typically consists of solid support, such as agarose or porous glass beads, immobilizing the affinity ligand. The mobile...
1.4K

You might also read

Related Articles

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

Sort by
Same author

Metal Hydroxide Composite with g-C<sub>3</sub>N<sub>4</sub> Catalyzed Oxidation of Antibiotics: A Step Towards Sustainable Development Goals for Antimicrobial Resistance.

Indian journal of microbiology·2026
Same author

Peptide Mapping Using Multienzyme Digestion Strategies Integrated with LC-HRMS Workflow: A Case Study.

Journal of peptide science : an official publication of the European Peptide Society·2026
Same author

A way forward for generic development of telmisartan tablets: Integrating critical material attributes and bio-predictive dissolution for target product profile.

European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V·2026
Same author

Tailoring isomeric transformations and surface adsorption dynamics of glycopeptides under thermal and photolytic stress.

Analytical and bioanalytical chemistry·2025
Same author

Proteomic Profiling Unraveling the Role of Lactate Dehydrogenase a in Vascular Repair and Functional Recovery after Spinal Cord Injury.

ACS chemical neuroscience·2025
Same author

Assessment of Thermal and Photolytic Stress Effects on the Stability of Primary Structure of Synthetic Liraglutide Using LC-HRMS/MS.

Journal of peptide science : an official publication of the European Peptide Society·2025

Related Experiment Video

Updated: Sep 26, 2025

Solid Phase Synthesis of a Functionalized Bis-Peptide Using "Safety Catch" Methodology
11:42

Solid Phase Synthesis of a Functionalized Bis-Peptide Using "Safety Catch" Methodology

Published on: May 15, 2012

24.8K

Synthetic pharmaceutical peptides characterization by chromatography principles and method development.

Nitish Sharma1, Divya Kukreja1, Tushar Giri1

  • 1The National Institute of Pharmaceutical Education and Research-Ahmedabad (Ministry of Chemicals and Fertilizers, Government of India), Opposite Air force Station, Gandhinagar, India.

Journal of Separation Science
|April 23, 2022
PubMed
Summary

This review covers peptide characterization and method development challenges. It details chromatographic techniques to ensure peptide product sameness and stability, crucial for pharmaceutical manufacturing.

Keywords:
high-performance liquid chromatographyion-exchange chromatographymethod developmentsize-exclusion chromatographysynthetic peptides

More Related Videos

Constructing Cyclic Peptides Using an On-Tether Sulfonium Center
07:11

Constructing Cyclic Peptides Using an On-Tether Sulfonium Center

Published on: September 28, 2022

2.8K
Synthesis and Characterization of 1,2-Dithiolane Modified Self-Assembling Peptides
09:54

Synthesis and Characterization of 1,2-Dithiolane Modified Self-Assembling Peptides

Published on: August 20, 2018

7.4K

Related Experiment Videos

Last Updated: Sep 26, 2025

Solid Phase Synthesis of a Functionalized Bis-Peptide Using "Safety Catch" Methodology
11:42

Solid Phase Synthesis of a Functionalized Bis-Peptide Using "Safety Catch" Methodology

Published on: May 15, 2012

24.8K
Constructing Cyclic Peptides Using an On-Tether Sulfonium Center
07:11

Constructing Cyclic Peptides Using an On-Tether Sulfonium Center

Published on: September 28, 2022

2.8K
Synthesis and Characterization of 1,2-Dithiolane Modified Self-Assembling Peptides
09:54

Synthesis and Characterization of 1,2-Dithiolane Modified Self-Assembling Peptides

Published on: August 20, 2018

7.4K

Area of Science:

  • Pharmaceutical Science
  • Analytical Chemistry
  • Biochemistry

Background:

  • Peptides, defined as polymers of 40 or fewer amino acids by the FDA, require rigorous characterization when manufacturing sources change.
  • Ensuring product sameness involves assessing impurities, degradation products, and aggregation levels of peptide active pharmaceutical ingredients.
  • Method development for peptide molecules presents unique challenges due to numerous variables impacting separation, recovery, and stability.

Purpose of the Study:

  • To review the fundamentals of peptide degradation and characterization methods.
  • To provide insights into chromatographic technique selection for peptide analysis.
  • To guide the optimization of method development parameters for peptide separation and recovery.

Main Methods:

  • Utilizes various chromatographic techniques including reversed-phase high-performance liquid chromatography-ultraviolet/high-resolution mass spectrometry, supercritical fluid chromatography, size-exclusion chromatography, and ion-exchange chromatography.
  • Focuses on method development strategies for peptide molecules.
  • Explores parameters influencing chromatographic separation, recovery, and stability.

Main Results:

  • Highlights the importance of chromatographic techniques in proving peptide product sameness.
  • Discusses common challenges in peptide method development.
  • Offers a guide for selecting appropriate chromatographic parameters for peptide characterization.

Conclusions:

  • Chromatographic techniques are essential for characterizing peptide active pharmaceutical ingredients and ensuring product consistency.
  • Careful consideration of method development variables is critical for successful peptide analysis.
  • This review provides a framework for optimizing peptide characterization through informed chromatographic parameter selection.