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Principles Of Column Chromatography01:13

Principles Of Column Chromatography

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

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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.
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Chromatography: Introduction01:10

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Chromatography is a technique used to separate compounds based on differences of partitioning between two phases, the stationary phase and the mobile phase.
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Chromatographic Methods: Terminology01:18

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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,...
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High-Performance Liquid Chromatography: Introduction01:11

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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.
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High-Performance Liquid Chromatography: Elution Process01:05

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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...
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Updated: Dec 19, 2025

Automated Hydrophobic Interaction Chromatography Column Selection for Use in Protein Purification
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Batch chromatography with recycle lag. II-Physical realization and experimental validation.

Abimaelle S Chibério1, Gonçalo F M Policarpo1, João C Antunes1

  • 1LAQV, REQUIMTE, Department of Chemistry, NOVA School of Science and Technology, Universidade NOVA de Lisboa, Caparica 2829-516, Portugal.

Journal of Chromatography. A
|June 8, 2020
PubMed
Summary

Researchers designed and validated an eluate recycling device (ERD) for batch chromatography. This system recycles eluate fractions, improving efficiency for large-scale applications.

Keywords:
Batch chromatography with recycle lagOne-column analogue of multicolumn chromatographyRecycle chromatography

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Area of Science:

  • Chemical Engineering
  • Analytical Chemistry
  • Separation Science

Background:

  • Batch chromatography is a key separation technique.
  • Recycle lag in batch chromatography can impact efficiency.
  • Optimizing eluate handling is crucial for large-scale applications.

Purpose of the Study:

  • To design, construct, and validate a prototype eluate recycling device (ERD).
  • To implement an approximate "first in, first out" (FIFO) system for eluate fractions.
  • To demonstrate the ERD's utility in large-scale batch chromatography.

Main Methods:

  • Detailed construction and setup of the ERD connected to a chromatography column.
  • Utilized two-way valves and six-port switching valves for operational control.
  • Experimental validation using reversed-phase chromatography for nucleoside mixture separation.

Main Results:

  • Successful experimental validation of the ERD setup.
  • Demonstrated effective recycling of eluate fractions.
  • Identified potential performance improvements through 3D printing of the fluid distributor.

Conclusions:

  • The developed ERD is a viable physical realization of batch chromatography with recycle lag.
  • The system is simple to set up, particularly for large-scale operations.
  • 3D printing offers a pathway to enhance ERD performance.