Continuous Purification from Flow Chemistry Systems with In-Line Flash Chromatography

Recorded June 15, 2022.

Join Christopher Thomson, Researcher at Heriot-Watt University, as he shares his work expanding the tool kit of automated flow synthesis with the development of in-line flash chromatography purification.

In this webinar, you will learn:

  • An introduction to flow chemistry and in-line purification technologies.
  • An overview of prior continuous chromatography methods, their advantages and limitations.
  • How to interface flow reactors with puriFlash automated chromatography systems.
  • ‘Tips & Tricks’ for performing continuous in-line flash chromatography.
  • Future prospects for developing in-line flash chromatography.

Chris completed his chemistry with biochemistry degree (MChem, 1st Class) at Heriot-Watt University, Edinburgh, in 2018 and was recipient of the ‘William H. Perkin Prize’ for excellence in organic chemistry. He was then awarded a PhD scholarship through the EPSRC funded centre for doctoral training in critical resource catalysis (CRITICAT), under the joint supervision of Dr Filipe Vilela and Dr Ai-Lan Lee.

Chris’ research focuses on the development and implementation of enabling technologies – especially flow chemistry – to enhance heterogeneous photocatalysis for organic synthesis.

He has published several papers on the development of flow systems featuring enabling technologies, such as: in-line NMR spectroscopy, in-line UV-Vis. spectroscopy, static mixing photocatalyst monoliths produced via functional material additive manufacturing, and most recently, reported the first example of continuous in-line flash chromatography – which will be the subject of this webinar.

Complement Flow Synthesis With In-Line Purification Using Flash Chromatography

In this application note, we describe a novel method to perform the continuous isolation of flow synthesis products from residual starting materials, catalysts or by-products to expedite chemical discovery. A promising new approach is highlighted here, featuring the Advion Interchim Scientific puriFlash® 5.250 preparative LC system.

The first results of our cooperation with the VilelaLAB and Continuum Flow Lab at Heriot-Watt University, Edinburgh, on this topic are outlined here and can be read in more detail in the following source: C.G.Thomson et al.: Expanding the Tool Kit of Automated Flow Synthesis: Development of In-line Flash Chromatography Purification, J. Org. Chem. 2021, 86, 20, 14079–14094.

Natural Products Purification – Improved Workflow with Mass Spec

Lab Manager Tech Trends Webinar. Recorded August 11, 2021.

This webinar features the Interchim PuriFlash® flash purification and preparative HPLC system and demonstrates separation of natural products, purification of specific target molecules and the combination of mass spectrometry detection of the same both for product confirmation and mass directed fraction collection using an Advion expression® compact mass spectrometer.

As a viewer, you will learn more about:

  • Natural product separation and analyte isolation from tea and herbs/spices starting from TLC separation
  • Column selection criteria for flash chromatography and preparative reverse phase HPLC
  • Integrating mass spectrometry detection of analytes from TLC plates and collected fractions
  • Mass spectrometry directed fraction collection to simplify the entire workflow and automate purification processes

Mass Directed Fraction Collection for Preparative SFC Collection of a 0.2% Impurity

This is one from a set of two application notes. In this application note, we determine the purity of collection of a small (0.2%) impurity in a concentrated sample mix by mass-directed fraction collection using supercritical fluid chromatography (SFC) coupled with the Advion expression Compact Mass Spectrometer (CMS).

For the other application note on the recovery and purification of two, three component standard mixtures, click here.

Advion co-authored with a major pharmaceutical partner to create two application notes highlighting the benefits of supercritical fluid chromatography (SFC) coupled to a compact mass spectrometer for mass directed fraction collection. These application notes conclude that mass‐directed fraction collection with a simplified passive splitter on the Preparative JASCO SFC coupled with expression CMS system increases purification selectivity by mass directed fraction collection while maintaining excellent recovery and purity. The MS accepted high concentrations of sample material with peak shapes trailing edges almost indistinguishable from the UV trace. Low, Middle and High isocratic flow rates all showed comparable MS source clearance leading to minimal tailing beyond the UV. Gradients further minimized peak tailing as expected on the UV and this was also seen on the TIC and XIC. Peak purities for all main band purifications were ≥99.9% pure and minor impurity collections showed equivalent success yielding 100% pure fractions. Acceptable recoveries for all collected fractions were 94% or better.

Peak Express – Unique Mass Spectrometer Software to Look Beyond the Noise and Detect Minor Components and Impurities for the expression® CMS

Introducing a new way to analyze and present mass spectral data, the Advion Peak Express patented software uses the delta spectrum (ΔS) to quickly and automatically detect significant peaks within mass spectra based on their relative change in intensity over time instead of their absolute intensity.

  • Find impurities and minor components that would otherwise be missed
  • Find unknowns in complex samples such as natural products
  • Automatically find adducts, dimers, fragments, side reactions, and other unexpected compounds in real time and post processing
  • Control mass-directed purification without providing an exact mass
  • Get extracted ion chromatogram (XIC) quality data while scanning the entire mass range

Mass Directed Fraction Collection

The Advion expression Compact Mass Spectrometer (CMS) was coupled to a Normal Phase Flash Chromatography system for mass directed fraction collection. Typically a UV detector is used to monitor compounds eluting from Normal Phase columns. A limitation of the UV detector is that it relies on compounds having a chromophore in the chemical structure, hence an analyte of interest may not be detectable if it lacks a suitable UV chromophore or coelutes with another compound. In addition, solvent systems can interfere with and hinder UV absorbance. The Advion expression CMS provides the ultimate in selectivity, unambiguously identifying residual starting materials, desired product ions, side reactants and impurities by their molecular weight despite co-eluting compounds and solvent peaks. Mass directed fraction collection greatly speeds up the chemist’s work flow with confidence in the content and purity of the collected fractions by eliminating off-line TLCs, workup and core lab LC/MS assays.

Simple, high-performance coupling of SFC to CMS for mass-directed fraction collection

This is one from a set of two application notes. In this application note, we review recovery and purity determination of two, three component standard mixtures by supercritical fluid chromatography (SFC) couped with the Advion expression Compact Mass Spectrometer (CMS).

For the other application note on the fraction collection of a 0.2% impurity, click here.

Advion co-authored with a major pharmaceutical partner to create two application notes highlighting the benefits of supercritical fluid chromatography (SFC) coupled to a compact mass spectrometer for mass directed fraction collection. These application notes conclude that mass‐directed fraction collection with a simplified passive splitter on the Preparative JASCO SFC coupled with expression CMS system increases purification selectivity by mass directed fraction collection while maintaining excellent recovery and purity. The MS accepted high concentrations of sample material with peak shapes trailing edges almost indistinguishable from the UV trace. Low, Middle and High isocratic flow rates all showed comparable MS source clearance leading to minimal tailing beyond the UV. Gradients further minimized peak tailing as expected on the UV and this was also seen on the TIC and XIC. Peak purities for all main band purifications were ≥99.9% pure and minor impurity collections showed equivalent success yielding 100% pure fractions. Acceptable recoveries for all collected fractions were 94% or better.