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Related Concept Videos

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...
Principles Of Column Chromatography01:13

Principles Of Column Chromatography

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...
Types Of Column Chromatography01:29

Types Of Column Chromatography

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
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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: 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: 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:

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Related Experiment Video

Updated: May 27, 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

Closed loop control of the multi-column solvent gradient purification process.

M Krättli1, G Ströhlein, L Aumann

  • 1Institute for Chemical and Bioengineering, ETH Zurich, Wolfgang-Pauli-Strasse 10, Zurich, Switzerland.

Journal of Chromatography. A
|November 8, 2011
PubMed
Summary

A new PID controller simplifies Multi-column Countercurrent Solvent Gradient Purification (MCSGP) operations using only UV signals. This automated system efficiently centers the product window for expected purity, aiding operators in continuous chromatography.

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Area of Science:

  • Chromatography
  • Process Control
  • Chemical Engineering

Background:

  • Multi-column Countercurrent Solvent Gradient Purification (MCSGP) is a continuous chromatographic technique.
  • Operator support is crucial for optimizing MCSGP performance and ensuring product quality.
  • Existing control methods may be complex or lack real-time adaptability.

Purpose of the Study:

  • To develop a Proportional-Integral-Derivative (PID) controller for assisting operators in MCSGP.
  • To implement a control strategy using only online UV signals for simplicity and speed.
  • To validate the controller's effectiveness in startup, steady-state maintenance, and disturbance rejection.

Main Methods:

  • Development of a PID controller utilizing online UV signals from each column outlet.
  • Operator-defined product window based on batch chromatogram analysis.
  • Testing the controller with a model protein and a peptide purification process.

Main Results:

  • The controller effectively drives the MCSGP unit to a stable steady state during startup.
  • It maintains a stable steady state and rejects external disturbances.
  • The controller ensures the withdrawn product window is centered within the expected purity region of the UV chromatogram.

Conclusions:

  • The developed PID controller is a reliable and efficient tool for MCSGP operation.
  • Simple control implementation using UV signals allows for fast control action.
  • The controller supports operators in achieving desired purity, complementing offline analytics.