Challenging separations, we all have faced this vexing problem. You synthesized your compound, analyzed it, and know your molecule is in there, based on LC-MS or TLC. Then, you do some method development using a silica TLC plate and see a major spot with some minor, early-eluting impurities. You think that the purification will be easy only to find that your “purified” compound has some co-eluting impurities. Now what? Should you change solvents or change stationary phase?

In this post, I will show how changing the column media but keeping the same solvents removed a co-eluting impurity in one of my reaction mixes.

Welcome to a second installment of the Biotage Cannabis Application development flash blog.  The first post, dated August 20  2016, outlined an orthogonal approach to isolating cannabinoids from winterized extract.  Give it read if you have not seen it

Today we investigate the “hot” topic of pesticide elimination from extract.  Specifically – Myclobutanil remediation.

Biotage®, a pioneer in Flash Purification, launched the unique, removable cap SNAP flash chromatography columns in 2007. This beneficial column design feature continues with the newest Biotage flash columns named Sfär columns.

Various flash chromatography sample loading options are available including liquid and dry loading. Choosing the right technique is important because your sample loading choices (sample solvent and dry load sorbent), can have a major impact on the results.

In this post, I compare the two techniques and show the benefits dry loading with a form of diatomaceous earth can bring to your purification.

For chemists, flash chromatography is part of their everyday synthesis workflow. For most syntheses, crude reaction mixtures are purified by normal-phase (aka adsorption) chromatography.  There are times, however, where the crude mixture’s complexity and polarity make normal-phase chromatography very challenging.  For these situations, reversed-phase (aka partition) chromatography may be a preferred option.

But, if you have only one flash system available, can you, should you, and how do you efficiently switch from non-polar, normal-phase solvents to polar, reversed-phase solvents – and back again without issues? In this post I'll attempt to shed some light on the topic.

Previously, I have posted on a normal-phase flash chromatography method to separate and isolate CBG from a CBD-rich hemp distillate. CBG is just one of many naturally occurring minor cannabinoids of interest in this fast-growing market.

The answer to this question is yes, reversed-phase can sometimes provide a better separation and thus better purification than normal-phase.  When is reversed-phase likely to be the better choice is a different, and likely better, question.

In this post I will try to demonstrate when reversed-phase is likely the better purification mode.

This is a question being asked of my colleagues and me more and more frequently, especially in pharma accounts.  Why?  Well, you are familiar with the adage – Time is Money, right.  Well this really applies to them. A new molecular entity (NME) created as a pharmaceutical can take up to a decade and a billion dollars to bring to market.  Granted, the biggest costs are in the clinical trials but the synthetic route and the time to discover and make the compound – and purify it – plays a major role within drug discovery and development. This timeline is not helped by the ever increasingly difficult-to-synthesize compounds being investigated as drug candidates today.

With that in mind, this post focuses on ways to speed the purification process without sacrificing purity and yield.

Recently, one of our readers wrote and asked how to determine solvent strength in normal-phase flash chromatography. This is an excellent question because solvent strength is one of several factors impacting flash chromatography performance.

In this post I will explain how solvent strength can easily be determined.

Media particle size and solvent flow rate play major roles in chromatographic separations including flash purification.  This is true in both reversed-phase chromatography (aka partition chromatography) as well as normal-phase chromatography. The roles played are related to the overall compound mass-transfer kinetics and diffusion/dispersion as they migrate through the column.  Smaller particles reduce sample dilution by reducing interstitial volume, while flow rate impacts the ability of molecules to efficiently pass through the porous particles. In this post, I will show how both particle size and flow rate impact flash chromatography.

In our more environmentally aware climate, chemical and pharmaceutical companies now prioritize reducing organic solvent use in chemistry labs. Employees and shareholders alike are pushing their companies to become greener which impacts how chemistry, both synthesis and purification, is performed.

Cannabis entrepreneurs continually seek to differentiate themselves from others in the market. Some focus on THC while others focus on CBD. What I have seen recently after attending some cannabis-specific conferences is a growing interest in isolating/purifying some of the minor, naturally occurring phytocannabinoids such as CBG and CBDV.

Varying the concentrations of mobile phase solvents during  flash purification chromatography enhances the ability of the technique to effectively isolate the desired compound from reaction by-products and unconsumed reagents.  Choosing how these concentrations will be varied over time has a significant effect on the purity and recovery of desired compounds.

What is the best starting strong solvent %?   What is the ending strong solvent %?  Should the mobile phase concentrations vary gradually in a linear manner or should they vary step-wise or something else altogether?  Most separations are performed once, occasionally a handful of times.  Because of this, spending effort optimizing a gradient is just not very productive unless there are aids in choosing the gradient profile that provides an effective purification with minimum effort.

Software in flash chromatography instruments, makes it simple to create a gradient.  Now, what should that gradient look like?

In this post I compare isocratic, step, and linear gradients and provide some sage advice on choosing among them.

If you synthesize organic amine compounds, especially heterocyclic, secondary, or tertiary amines, you likely have encountered problems with their chromatography using silica columns. With the amine groups being basic and silica being acidic, there is a natural attraction between the two. This sometimes strong attraction often requires the use of a competing amine in the solvent system. Modification of the mobile phase with the addition of a solvent like triethyl amine can provide a successful purification. Often times the use of an amine-modified stationary phase can provide the needed conditions to avoid the acid-base interaction that can interfere with a successful flash chromatography purification.

In this post I will discuss how amine-functionalized silica can simplify organic amine purification and your life (at least in the lab).

In many of my previous posts I have used the term column volume, typically abbreviated as CV, as a value used to help determine separation quality and loading capacity.  However, I recently was asked a question about this topic from a chemist who understands the column volume concept but wanted to better understand its definition and how it is determined.

In this post I will explain what a column volume is and how it is determined empirically.

This is an age-old question that has been around a long time, perhaps as long as me (and I have been around a while) –  “Does silica dissolve in methanol?”