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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:
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.
Size-Exclusion Chromatography01:08

Size-Exclusion Chromatography

In size-exclusion chromatography (SEC), also known as molecular-exclusion or gel-permeation chromatography, molecules are separated based on their sizes. This technique is important for separating large molecules such as polymers and biomolecules. The two classes of micron-sized stationary phases encountered in SEC are silica particles and cross-linked polymer resin beads. Both materials are porous, but their pore sizes vary significantly.
Silica particles offer advantages such as rigidity,...
Ion Exchange01:17

Ion Exchange

Ion exchange chromatography separates charged molecules from a solution by reversibly exchanging them with mobile, or 'active', ions associated with the oppositely charged stationary phase. This method can be used to separate ions, soften and deionize water, and purify solutions. The polymers comprising the ion-exchange column are high-molecular-weight and chemically stable polymers, crosslinked to be porous and essentially insoluble. They are also functionalized with either acidic or basic...
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...

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Related Experiment Video

Updated: Jun 25, 2026

Fabrication of a Dipole-assisted Solid Phase Extraction Microchip for Trace Metal Analysis in Water Samples
09:42

Fabrication of a Dipole-assisted Solid Phase Extraction Microchip for Trace Metal Analysis in Water Samples

Published on: August 7, 2016

Polymer microchips integrating solid-phase extraction and high-performance liquid chromatography using reversed-phase

Jikun Liu1, Chien-Fu Chen, Chia-Wen Tsao

  • 1Department of Mechanical Engineering, University of Maryland, College Park, Maryland 20742, USA.

Analytical Chemistry
|March 10, 2009
PubMed
Summary
This summary is machine-generated.

This study introduces polymer microfluidic chips for peptide separation using polymethacrylate monoliths. The integrated design enhances sensitivity and separation performance for high-performance liquid chromatography (HPLC).

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Fabrication of the Thermoplastic Microfluidic Channels
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Fabrication of the Thermoplastic Microfluidic Channels
16:00

Fabrication of the Thermoplastic Microfluidic Channels

Published on: February 3, 2008

Area of Science:

  • Analytical Chemistry
  • Materials Science
  • Biotechnology

Background:

  • Polymer microfluidic chips offer advantages for miniaturized analytical systems.
  • High-performance liquid chromatography (HPLC) is crucial for peptide separation.
  • Integrating sample preparation with separation enhances analytical efficiency.

Purpose of the Study:

  • To develop and demonstrate a polymer microfluidic chip for integrated peptide separation and sample preparation.
  • To improve sensitivity and separation performance in liquid chromatography.
  • To enable high-pressure operation for efficient separations.

Main Methods:

  • In situ photopolymerization of polymethacrylate monoliths within microfluidic chips.
  • Integration of a solid-phase extraction (SPE) element with a reversed-phase separation column.
  • Utilizing solvent bonding and a high-pressure needle interface for chip integrity.
  • Performing gradient reversed-phase separation on fluorescein-labeled peptides and BSA tryptic digest.

Main Results:

  • Demonstrated efficient peptide separation and sample enrichment on-chip.
  • Achieved a 150-fold improvement in detection sensitivity.
  • Reduced peak width by 10-fold for enhanced separation resolution.
  • Successfully operated the polymer chips at pressures exceeding 20 MPa.

Conclusions:

  • The developed microfluidic HPLC system provides a powerful platform for sensitive peptide analysis.
  • On-chip SPE and separation integration significantly boost analytical performance.
  • The robust chip design supports high-pressure, efficient liquid chromatography separations.