UV-Based Fraction Collection
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UV-Based Fraction Collection

 

Intro and Background Literature

Reversed-phase liquid chromatography (LC) is often associated with quantitative or analytical-scale analysis, but it is also useful as a preparative scale tool for purification. For any purification project, there are three possible targets: purity, yield, and throughput. These targets are related to one another. Two of these targets can be achieved at the expense of the third. You can clarify your path forward by prioritizing the objectives of your purification project before method development.

Tips for bulk purification

Bulk purifications prioritize purity and yield at the expense of throughput, and these guidelines will assist you with method development.

  • Your column only needs to resolve the target compound from impurities. All other peaks can coelute

  • Always verify that your sample is soluble in mobile phase solvents.

  • Work out the method and loading conditions on the analytical system before scaling them to the preparative system.

  • To account for dwell volume differences, remember to incorporate an isocratic hold where necessary

More Information

Principles and Practical Aspects of Preparative Liquid Chromatography - A Primer

A Tale of Two Samples: Understanding the Purification Workflow from Different Perspectives, Part 1

A Tale of Two Samples: Understanding the Purification Workflow from Different Perspectives, Part 2

Agilent InfinityLab LC Purification Solutions - Quick Reference Guide

Agilent InfinityLab LC Purification Solutions - Purify Your Samples with Maximum Flexibility

1290/1260 LC/MSD System, preparative or analytical-scale

Fraction collection workflow

Set up parameters, target compounds, and fraction collection triggers. See Fraction Collectors.

Main steps in a Fraction collection workflow

  1. Set up a method with fraction collection trigger signals. See Create a new method.

  2. Prepare acquisition, submit samples

  3. Evaluate results Review the fraction results in Data Analysis

  4. Report the fraction results: Fraction Collection report items

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