How does solvent choice impact flash column chromatography performance?

Selectivity and solvent strength are the most important factors that determine success or failure of a chromatographic separation. These two independent dynamics apply to both normal- and reversed-phase chromatography and should be optimized, especially when high fraction purity is needed.

In this post I will discuss the impact that elution solvent choice has on both normal- and reversed-phase purification. Continue reading How does solvent choice impact flash column chromatography performance?

How do I determine loading capacity in reverse phase flash column chromatography?

As the popularity of prep-scale, reversed-phase flash chromatography increases, so does the frequency that I get asked this question, “How do I determine loading capacity in reversed-phase flash chromatography?”

In the world of HPLC, loading capacity isn’t normally a concern as it is primarily an analytical technique.  In the synthetic organic chemistry world, most purification is performed with silica gel where flash column purification methods are developed and loading capacity estimated from TLC data.  However, when normal-phase flash does not work and reversed-phase flash is needed, the question of how to determine reversed-phase loading capacity comes up.

In this post I will attempt to provide some guidelines to help you understand and determine reversed-phase flash chromatography loading capacity. Continue reading How do I determine loading capacity in reverse phase flash column chromatography?

Flash column equilibration – is it a waste of time or necessary step?

Flash column chromatography has been practiced by chemists since the 1970s. That practice required the silica in the column be properly wetted to remove trapped gasses to ensure uniform flow (remember those days of not letting air into the column?). Today, with automated flash chromatography systems and pre-packed cartridges as the norm, chemists ask me – do I really need to pre-equilibrate my cartridge?

In this post I explore the impact on chromatography that equilibration, and lack thereof, has on separation performance.

Continue reading Flash column equilibration – is it a waste of time or necessary step?

How do I create an efficient gradient flash chromatography method?

For most organic and medicinal chemists flash chromatography is just another step in the synthesis work flow – react, analyze, purify, react, analyze, purify… until the final product is made. The desired product of each reaction, and the mixture of other species present are, of course, different with each cycle.  Separating the desired compound efficiently without a lot of hassle is something I have written about in this post as well as in others in this series.

In this post, I’ve written about how that TLC (thin layer chromatography) plate you use for monitoring your reaction can be used to create reliable, efficient, effective gradients.

Continue reading How do I create an efficient gradient flash chromatography method?

How do I Choose the Right Column Size for Purification by Flash Chromatography?

In all my years of working with medicinal and organic chemists, I have found that choosing how many grams of silica to use for purification by flash chromatography is something frequently guessed at. Getting the size of the column right is awfully important because using too few grams of silica will doom your purification to failure and using more an optimal mass of the stationary phase means the purification  consumes excess silica, solvents, and a chemist’s time. To determine the optimal amount of silica for a purification, I rely on a factor called ΔCV (delta Column Volume) to identify the best loading capacity on any cartridge. I have also found that ΔCV this is a better loading capacity predictor for flash purification than ΔRf.

Continue reading How do I Choose the Right Column Size for Purification by Flash Chromatography?

How to Optimize TLC to Enhance Purification by Flash Chromatography

In this article I discuss the optimization of solvent ratios to generate ideal Rf (retention factor) values on TLC plates.  Then I show how maximizing efficiency of flash chromatography achieves higher loading with rapid and reliable isolation of compounds, reduced solvent use and improved separation. Continue reading How to Optimize TLC to Enhance Purification by Flash Chromatography

Using TLC to Scout Flash Chromatography Solvents

TLC is the tool most used for normal-phase flash chromatography method development. For many chemists, a solvent system of hexane (or heptane) + ethyl acetate is the first, and sometimes only, solvent system evaluated. Though often useful, ethyl acetate may not always provide the optimal purification conditions. Continue reading Using TLC to Scout Flash Chromatography Solvents