Knowing the Chromatography Workflow Consisting of many different and evolving techniques, chromatography is used across multiple branches of science to separate molecules. With a focus on preparation and/or analysis of these molecules, there are two main phases of chromatography used for separating components: the mobile phase and the stationary phase. For glassware, labware and life science benchtop equipment suggestions by chromatography phase, please see the following pages.
Preparation
When preparing the column, careful thought must be given to provide best sample separation. Consider sample polarity, size, and affinity. Preparation is dictated by what type of chromatography is used – analytical or preparative.
Running the Column
Columns must be set up properly for samples to run effectively. Factors like loading the column or making sure it is vertically straight, could make a vast difference in sample collection.
Fraction Collection
Fraction collection varies, depending on how well you can visualize your sample on the column. Best practice dictates to collect many small fractions which can be pooled together once their purity is confirmed.
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Analysis
Thin layer chromatography is a common technique to confirm which fraction contains the desired eluent. Additional techniques like UV/Vis spectrophotometry can be used separately or in concert.
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Concentration
Often, the pure sample must be at a higher concentration than what the pooled fractions permit. Tools such as evaporators are available.
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Clean Up
Properly cleaned glassware and plasticware allows for confidence in future analyses using the same methodology.
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