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?”
Equilibrating silica flash chromatography columns is something I always do. There are chemists who see this as an unnecessary, time-and-solvent-wasting step. However, because getting consistent, predictable results is a priority for me, I equilibrate to remove the variability that can be caused by heat generated as solvent initially contacts the silica. Consistency is really important when running flash column chromatography because re-runs are time consuming and may put your compound at risk.
In this post, I examine the role of equilibration speed and duration to show its impact, or lack there of, on purification performance.
Reversed-phase flash chromatography usage is increasing rapidly. In fact, over the past 10 or so years, reversed-phase flash chromatography use has increased a dramatic 650%! This is amazing growth despite the fact that reversed-phase flash columns are considerably more expensive than silica columns and you need to evaporate water from your fractions. So, what’s driving this change in chemists’ modus operandi?
In this post, I will explain why chemists are increasingly using reversed-phase flash chromatography for routine, intermediate, and final compound purification and provide and example as well.
Most flash column manufacturers now offer “high performance” flash chromatography columns with the promise of higher loading, increased purity, and even reduced solvent consumption. Working for Biotage, I have made those valid claims for our products as well.
On July 26th, 2018, Bob Bickler, Senior Technical Specialist, recorded a webinar on A Roadmap to Successful Flash Chromatography. To learn more, read the description below as well as watch the recording!
For most synthesis and natural product chemists, flash chromatography is the primary tool for purification and isolation of compounds of interest. Purification methods include flash system defaulted linear gradients (e.g. 0-100%), active gradient modification (on-the-fly) during purification, and unique method creation based one either the chemist’s experience or TLC data (typically a linear gradient). Rarely do chemists work to optimize the purification to maximize target compound purity by employing a step gradient. In this post, I discuss the value optimized step gradients provide chemists.