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.
Purifying polar organic compounds can be very challenging. In a previous post I have discussed using reversed-phase flash chromatography to retain and purify ionizable and ionic compounds. My colleague, Dr. Elizabeth Denton, has also posted a blog on purifying very polar peptides as well. Sometimes, however, despite all your efforts with reversed-phase, success is elusive. When this happens, what do you do?
"With using the entire workflow, we're able to do things more efficiently," stated Dr. Justin Anglin during our interview in February at the Center for Drug Discovery at Baylor College of Medicine. Keep reading to find out more about how his role as the Director helped influence their state-of-the-art lab to include a full suite of Biotage products.
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.
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.
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.
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.
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).
This is an interesting question that does not have a straightforward answer. In fact, there are many materials that are potentially useful sorbents for dry loading crude mixtures. Some of the more popular are silica, diatomaceous earth (e.g. ISOLUTE® HM-N, Celite®), alumina, and Florisil®. The sorbent choice can influence your purification results because each of the available media have different chemistry and capacity. In most cases, sample/sorbent reactivity really is not a major concern, though it can occur. What is important is the sorbent’s capacity to adsorb/absorb all of your sample and the ratio of your crude sample to the amount of dry load sorbent.
Dry loading crude samples for flash purification typically works better than liquid loading, especially for challenging purifications. In this post, I discuss how the ratio of crude sample to dry load sorbent impacts purification performance.
Learn more about how to get in control of your Flash Purification.With Biotage® Selekt and Sfär chemists can be in full control of Flash Purification.
UV detection and fractionation is ubiquitous in flash chromatography. It is the default methodology used to detect and collect eluting compounds. Today’s flash chromatography systems offer UV-triggered fractionation on one, two, or a range of wavelengths in order to either increase fractionation specificity, yield, or increase sensitivity.
On December 6th, 2018, Bob Bickler, Senior Technical Specialist, recorded a webinar on Inspiring Productivity with Modern Flash Chromatography. To learn more, read the description below as well as watch the recording!