Successfully completing an organic synthesis involves three typically lengthy, but crucial steps: reaction monitoring, compound identification, and purification. These steps can create bottlenecks in the synthesis, unless a purpose-built, streamlined, and reliable workflow is on hand.
This whitepaper delves into some of the challenges faced by synthetic organic chemists as they navigate compound identification and purification struggles. It also describes how using a workflow that couples thin layer chromatography (TLC) and mass spectrometry (MS) to flash purification can help increase the efficiency of the entire process.
Key Objectives：
- Understand how a workflow that combines thin-layer chromatography, mass spectrometry, and flash purification can increase the speed and efficiency of an entire organic synthesis process.
- Learn about prep-free techniques that have become the new standard: scraping TLC spots, developing flash methods and performing long liquid chromatography/MS runs are not necessary with modern instrumentation that is specifically developed to take the prep-work and guess-work out of the purification process.
- Become informed about a cost-effective, user-friendly benchtop workflow solution for organic compound identification and purification that can save time and money for any lab.
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.
The combination of the Advion Interchim Scientific expression® CMS and puriFlash® flash chromatography technologies speed up the synthesis, purification and characterization of a compound.
Using the Advion Interchim Scientific expression® Compact Mass Spectrometer, the Plate Express™ TLC Plate reader, the ASAP® Atmospheric Solids Analysis Probe, and the puriFlash® XS520 flash purification system, it is now possible to execute a fast, user-friendly workflow to run TLC, harness the power of your cell phone to develop a flash method, and finally utilize a simple dipping probe to confirm fractions in seconds.
With easy sampling techniques, intuitive software, and advanced detection technology, you can easily and confidently synthesize, purify and characterize your compound, in a flash.
The compound synthesis, purification and confirmation process often presents challenges throughout. From ensuring successful synthesis of your product, setting up a flash purification method, and finally confirming ID of fractions, the lengthy work flow can be time consuming and repetitive.
In this application note, we demonstrate how the combination of Advion Interchim Scientific technologies speed up the synthesis, purification and characterization of a compound.
This series features the Advion Interchim Scientific expression® Compact Mass Spectrometer (CMS), the Plate Express™ TLC Plate Reader, the Atmospheric Solids Analysis Probe (ASAP®), and the puriFlash® XS520 flash purification system.
The Advion Interchim Scientific suite of systems and consumables allows users to harness the power of mass spectrometry, flash chromatography, prep LC and more. Simple and robust, download our brochure to learn more about:
- The Advion Interchim Scientific expression® compact mass spectrometer (CMS): A fast and easy analytical tool for the organic chemist. Ideal for fast reaction monitoring, the expression® CMS features a single quadrupole that can adapt to multiple ionization sources in seconds, including both ESI and APCI. The expression® CMS offers a variety of novel sampling techniques, including fast assay methods for liquids, solids, gases, and even air-sensitive compounds.
- Direct mass analysis of TLC plates in 30 seconds at the push of a button with Plate Express™
- One-touch analysis of solids and liquid samples with the ASAP® probe
- LC/CMS
- puriFlash® ultra performance flash purification: Ideal for method development and purification of rare and high added value compounds, the Interchim family of puriFlash® systems offer users a wide range of throughput options and the highest recovery rates at >95%.
- Mass-Guided Purification: the Interchim puriFlash® + Advion CMS offers the ideal solution for Flash-MS, and can provide fraction identification in <30 seconds.
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Fill out the form to download the full Advion Interchim Scientific brochure now.
Merck & Co.
Abstract
The evolution of supercritical fluid chromatography (SFC) instrumentation, improved detection capability, and expanded modifier range has led to extending the reach of SFC to the analysis of a broader spectrum of analytes beyond enantioselective separations. However, preparative SFC has yet to see the same technological revitalization, especially in regards to the purification of highly polar analytes. Enhanced fluidity liquid chromatography (EFLC) has been demonstrated as one of the ways to extend the applicable range of SFC instrumentation to highly polar analytes such as proteins, carbohydrates, and nucleotides. Despite recent applications of EFLC for challenging mixtures of hydrophilic metabolites and analogs, its viability in preparative purification, which is of great importance to the pharmaceutical industry, remains unknown. Herein, multiple chromatographic parameters that are critical to achieve feasible EFLC purification methods were investigated, including system pressure as a function of modifier composition (for several MeOH:H2O ratios), effect of diluent injection conditions on peak shape, and optimization of mass load with diluent composition. The usage of 50% acetonitrile or methanol diluents provided the most volumetric loading capacity. In the case of sucrose, leveraging higher analyte solubility in water proved to be more favorable than the volumetric loading capacity of diluents with higher organic content. In fact, an 80 mg injection of sucrose was possible on a 2 cm preparative HILIC column with minimal peak shape degradation. The combined information led to the successful demonstration of EFLC for the preparative separation of sugars using readily available MS-directed SFC instrumentation.
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.
