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.
Justus Liebig University Giessen, University of Modena and Reggio Emilia
Abstract
Effect-directed analysis (EDA) by the combination of high-performance thin-layer chromatography (HPTLC) with biologi- cal and enzymatic assays represents one of the latest tools available for the rapid bioprofiling of complex matrices, such as plant extracts. In this ambit, the aim of this project was the non-targeted screening of inflorescence extracts from ten different hemp varieties for components exhibiting radical scavenging, antibacterial, enzyme inhibiting and estrogen-like effects.
The characterization of two prominently multipotent bioactive com- pound zones was finally achieved by HPTLC-HRMS and preliminary assigned as cannabidiolic acid and cannabidivarinic acid.
HTPLC analysis was coupled via the Advion Plate Express® TLC Plate Reader.
The compound synthesis, purification and confirmation processes often present challenges. 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.
This webinar will provide a foundation for flash purification processes, and will showcase a novel work flow concept that breaks down the process in to easy steps for optimal success. We will offer tips to optimize each essential segment of the workflow, and share seamless steps to quickly and easily drive the process from reaction monitoring to flash purification and fraction ID.
Presented by Dr. Daniel Eikel, Director of Customer Service and Product Applications, Advion.
This webinar was hosted by C&EN and recorded 8/13/20.
In this study, total saponins were extracted from Pleurotus ostreatus cultivated with Astragalus as one of organic culture substrates. High Performance Thin Layer Chromatography (HPTLC) assay showed total saponins could be separated effectively, and four kinds of spots were identified as AG I, AG II, AG III, and AG IV, respectively. FTIR spectra based on HPTLC separation assay showed the saponin characteristic groups including ‐OH, C‐H, C=O, and the glycoside linkaged to sapogenin group C‐O‐C, suggesting the four kinds of spots belonged to cycloartane‐type triterpene saponins. The primary mass spectra of precursor ion (HPTLC‐ESI‐MS) assay further proved the main composition of four kinds of spots was AG I‐IV, respectively. Physical properties, including the detection of specific rotation and melting point, revealed the separation of high‐purity saponin monomer by HPTLC. HPTLC‐dual wavelength spectrodensitometric method detection showed that content of astragaloside I‐IV was ranged from 0.2 to 0.5 mg/g, and the total astragalosides contents attained to 1.397 mg/g, indicating P. ostreatus could bioaccumulate astragalosides from Astragalus. These results demonstrated the characterization of astragalosides based on the separation of HPTLC was effective, and supported to consider astragalosides‐enriched P. ostreatus as functional edible fungus for food and medical applications.
Practical Application
Currently, the consumption of enriched foods has become common and continues to increase due to urgent demanding for foods with high nutritional value. Pleurotus ostreatus is a functional edible fungus, which not only can produce secondary metabolites, but can enrich bioactive ingredients. Astragalosides have a wide range of biological activities, especially currently being tested as cardioprotective agent. In this study, P. ostreatus was cultivated through adding Astragalus into culture substrates, which realized massive enrichment of astragalosides. Astragalosides‐enriched P. ostreatus as functional edible fungus could be extensively used in food and medical areas, especially for the prevention of cardiovascular diseases.
Analysis was performed by LC/MS using the Advion expression Compact Mass Spectrometer (CMS).
Mass spectrometry is an analytical technique used to determine the mass-to-charge ratio of ions in a sample and therefore the sample composition. This method is used across many industries including food and beverage, environmental monitoring, and clinical. There are many features of mass spectrometers that must be considered before purchasing such as the required mass analyzer technology, ionization source, and dissociation technique. Join Lab Manager and our panel of experts as we discuss how to decipher mass spectrometry technology offerings and find the right solution for your lab.
As an attendee, you will learn more about:
Available technology for performing mass spectrometry
Features to consider when purchasing mass spectrometry instruments
How to find the right mass spectrometer for your application
Environmental monitoring is vital to understanding the world around us. Air quality, water, and soil testing all provide valuable information about an environment’s status. Tools and technologies for environmental monitoring can be used to evaluate the sanitation of a food and beverage lab or to monitor an ecosystem’s recovery after an oil spill. Join Lab Manager and our panel of experts as we review the tools available for environmental testing and monitoring.
As an attendee, you will learn more about:
Developments in technology for environmental testing and monitoring
Considerations when purchasing technology for environmental testing and monitoring
Applications for environmental testing and monitoring across industries
This webinar was hosted by Lab Manager Tech Trends and recorded July 9, 2020.
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 collection of solid particulates and liquids from surfaces by the use of cloth swipes is fairly ubiquitous. In such methods, there is a continuous concern regarding the ability to locate and quantitatively sample the analyte species from the material. In this effort, we demonstrate the initial coupling of an Advion Plate Express plate reader to a liquid sampling–atmospheric pressure glow discharge (LS-APGD) microplasma ionization source with an Orbitrap mass spectrometer to perform uranium isotopic analyses of solution residues on cotton swipes. The Plate Express employs a sampling probe head to engage and seal against the swipe surface. Subsequentially, the analyte residues are desorbed and transported within a 2% HNO3 electrolyte flow to the ionization source. Quantitative recoveries were observed following a single 30 s extraction step, with the absolute mass sampled per extraction being ∼100 ng. While the intrasample variability in the analytical responses for triplicate sampling of the same swipe yield ∼30% RSD, this lack of precision is offset by the ability to determine isotope ratios for enriched uranium specimens with a precision of better than 10% RSD. Pooled, intersample precision (n = 9) was found to be <5%RSD across the various sample compositions. Finally, 235U/238U determinations (ranging from 0.053 to 1.806) were accurate with errors of <10%, absolute. The 234U- and 236U-inclusive ratios were determined with similar accuracy in enriched samples. While the driving force for the effort is in the realm of nuclear nonproliferation efforts, the ubiquitous use of cloth swipes across many application areas could benefit from this convenient approach, including the use of versatile, reduced-format mass spectrometer systems.
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 combination of the Advion Interchim Scientific® expression® CMS puriFlash® flash chromatography technologies speed up the synthesis, purification and characterization of a compound.
