Learn how SpiroChem AG (Switzerland) utilized the Advion expression® Compact Mass Spectrometer (CMS) and Plate Express™ TLC Plate Reader for reaction monitoring and compound identification to aide in accelerating drug discovery.
Abstract:
In highly competitive research environments, the ability to access more complex structural spaces efficiently is a predictor of a company’s ability to generate novel IP-protected small molecule candidates with adequate properties, hence filling their development pipelines. SpiroChem is consistently developing new synthetic methodologies and strategies to access complex molecular structure, thereby facilitating and accelerating small molecule drug discovery. Pushing the limits of what are perceived as complex molecular structures allows SpiroChem and its clients to unleash creativity and explore meaningful chemical spaces, which are under-exploited sources of novel active molecules. In this article, we [SpiroChem] explain how we differentiated ourselves in a globalized R&D environment and we provide several snapshots of how efficient methodologies can generate complex structures, rapidly.
A: The Rappsilber Laboratory is interested in studying the machinery of life, in particular protein structures and their interactions in living cells. Determining protein behaviour is crucial for developing new drugs or understanding terrible diseases such as cancer. We are doing this by developing state-of-the-art methods and new chemical tools for crosslinking mass spectrometry (crosslinking-MS), including new crosslinker reagents and crosslinking chemistry. Working on the chemistry-biology interface, our work encompasses chemical synthesis of peptides, fluorophore tagged sensors, peptide- and protein-reactive crosslinking reagents and bioorthogonal metal catalysts.
Q: WHAT WAS YOUR PREVIOUS WORKFLOW OR CHALLENGES?
A: This year (2020) we built a chemistry lab from scratch, here at the Institute of Biotechnology (TU Berlin). We had the opportunity to really think about our work flow requirements and create the most efficient chemical synthesis pipeline for a modern, 21st century, small chemistry laboratory. However, we are a satellite campus, located more than 10 km from the Institute of Chemistry, which meant that we were constrained by the lack of usual chemistry infrastructure. One of the major challenges that we faced here, is that without chemistry-specific analytical capability, we are working almost blind. Every step of chemical synthesis, including reaction monitoring, work-up, purification and final product characterisation required an exhausting and disruptive journey across Berlin!
Q: WHY DID YOU INCORPORATE THE EXPRESSION® CMS INTO YOUR LABORATORY?
A: We operate an advanced proteomics facility, with cutting edge, high-precision, high sensitivity mass spectrometers for analysing extremely complex peptide mixtures. But a chemist needs a workhorse, not a Formula 1 car. The Advion Interchim Scientific expression® CMS was a no-brainer. We needed a system that is versatile, compact, easy-to-use, easy-to-maintain (~10 minute calibration), robust and allows fast analysis (result in <30 seconds). This system is all of this, but furthermore it is a delight to use. The ASAP® Direct Analysis Probe works fantastically well for analysis of solids and liquids. Switching between APCI and ESI is so quick and easy. The Plate Express® system makes it a joy to identify reaction products directly from TLC plates, greatly simplifying subsequent purifications. The Plate Express™ also works really well for identifying synthesized peptides from TLC spots. The expression® CMS sits at the heart of our chemistry lab and workflow.
Q: WHO WOULD YOU RECOMMEND TO PURCHASE THE EXPRESSION® CMS?
A: We would recommend the Advion Interchim Scientific expression® CMS to literally any research laboratory, big or small. The versatility of the system is truly impressive.
A: The research of the new Institute for Drug Discovery seeks to combine computational and experimental efforts to investigate proteins and their interactions with small molecule substrates. The Medicinal Synthetic Chemistry Core is part of the new institute and responsible for the synthesis of potentially bio-active compounds based on the computational results predicting a high binding affinity to the proteins’ active sites.
Q: WHAT WAS YOUR PREVIOUS WORKFLOW OR CHALLENGES?
A: Previously, we separated all unknown by-products of our synthesis in order to investigate their structure based on NMR and external mass spec service. This time wasting workflow often resulted in a dead-end strategy with low efficiency.
Q: WHY DID YOU INCORPORATE THE expression® CMS INTO YOUR LABORATORY?
A: Since the purchase of the CMS, we improved our strategic synthesis planning extraordinarily. By using Plate Express™ as TLC-Interface together with the expression® CMS, the by-products could become quickly separated on TLC-plates and directly analyzed by mass spectrometry afterwards. Now we see it, now we know it, and we get it quickly purified.
Q: WHO WOULD YOU RECOMMEND TO PURCHASE THE expression® CMS?
A: I will highly recommend the CMS to all research institutes with a strong synthetical and natural product background of small molecules.
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.
Rey Juan Carlos University, Complutense University of Madrid, University Rennes
Abstract
Lichens produce unique secondary metabolites with a rich potential as bioactive compounds. In many cases, the use of these molecules is limited by the low concentration of these compounds in thalli, low growth rate in culture, and changes in chemical patterns between thalli and aposymbiotic culture. In addition, the massive collection of some species of industrial interest can cause damage to lichen diversity and the associated environment. Six lichenized fungi (Arctoparmelia centrifuga, Parmelia saxatilis, Parmelina tiliacea, Platismatia glauca, Xanthoparmelia tinctina, and Usnea ghattensis) with biotechnological interest and belonging to Parmeliaceae have been cultured in order to test culture conditions and obtain enough biomass for further studies. In addition, we analyzed the compounds synthetized in axenic conditions and they were compared with chemosyndromes identified in complete thalli. Arctoparmelia centrifuga, P. saxatilis, P. tiliacea and X. tinctina were successfully cultivated while for P. glauca and U. ghattensis we only obtained sporulation and germination of the spores. The chemical pattern of the compounds secreted into the culture media varied significantly from the chemosyndrome of the whole thallus. Phenolic compounds of pharmacological and industrial interest (usnic acid, aspicilin, α-alectoronic acid, physodic acid, lobaric acid and nordivaricatic acid) and a wide variety of potentially bioactive compounds were obtained during the culture process.
Analysis was performed by LC/MS using the Advion Interchim Scientific® expression® Compact Mass Spectrometer (CMS).
Cornell University, Kaiser Permanente Fontana, Bar-Ilan University
Abstract
The consumption of teff (Eragrostis tef), a gluten-free cereal grain, has increased due to its dense nutrient composition including complex carbohydrates, unsaturated fatty acids, trace minerals (especially Fe), and phytochemicals. This study utilized the clinically-validated Gallus gallus intra amniotic feeding model to assess the effects of intra-amniotic administration of teff extracts versus controls using seven groups: (1) non-injected; (2) 18Ω H2O injected; (3) 5% inulin; (4) teff extract 1%; (5) teff extract 2.5%; (6) teff extract 5%; and (7) teff extract 7.5%. The treatment groups were compared to each other and to controls. Our data demonstrated a significant improvement in hepatic iron (Fe) and zinc (Zn) concentration and LA:DGLA ratio without concomitant serum concentration changes, up-regulation of various Fe and Zn brush border membrane proteins, and beneficial morphological changes to duodenal villi and goblet cells. No significant taxonomic alterations were observed using 16S rRNA sequencing of the cecal microbiota. Several important bacterial metabolic pathways were differentially enriched in the teff group, likely due to teff’s high relative fiber concentration, demonstrating an important bacterial-host interaction that contributed to improvements in the physiological status of Fe and Zn. Therefore, teff appeared to represent a promising staple food crop and should be further evaluated.
Analysis was performed by LC/MS using the Advion Interchim Scientific expression® Compact Mass Spectrometer (CMS).
This study evaluates the effects of intra-amniotic administration of yacon (Smallanthus sonchifolius) flour soluble extracts (YFSE) on intestinal bacterial populations, brush border membrane (BBM) functionality and morphology, by using the Gallus gallus model.
Highlights:
Yacon flour soluble extract (YFSE) increased the relative abundance of bifidobacterial and Lactobacillus.
YFSE upregulated the expression of iron and zinc transporters.
YFSE decreased gene expression of inflammation marker and hepcidin.
YFSE increased glycogen concentration in pectoral muscle.
YFSE improved BBM functionality and intestinal morphology.
Analysis was performed by LC/MS using the Advion expression Compact Mass Spectrometer (CMS).
Selcuk University, University “G. d’Annunzio” of Chieti-Pescara, University of Perugia, University of Urbino
Abstract
Coronilla species, belonging to the Coronilla genus (Fabaceae), have long been used in traditional medicine for treating cold, diabetes, pain, and as cardiotonics. The goal of the present study was to explore the phytochemical composition and pharmaco-toxicological properties of C. minima. In this regard, phenolic content, scavenging/reducing properties and antimicrobial activity toward pathogen bacterial (Escherichia coli, Pseudomonas aeruginosa, Bacillus cereus, Staphylococcus aureus) and fungal strains (Candida albicans, C. tropicalis, Aspergillus tubigensis and A. minutus) were investigated. Extract effects on human colon cancer HCT116 cell viability were also assayed. Finally, a bioinformatics approach was conducted with the aim to identify putative microbial and human protein targets underlying antibacterial, antimycotic, and antiproliferative effects.
Analysis was performed by LC/MS using the Advion expression Compact Mass Spectrometer (CMS).
New 1-hydroxy-10-alkylacridin-9(10H)-one BF2-chelates absorb in the blue-green part of the electromagnetic spectrum and emit fluorescence with moderate quantum yields of 8–45% in toluene. The dyes show large Stokes shifts about 4300 cm−1, decay times between 5 ns and 15 ns in toluene and high photostabilities. Introduction of a fluorine atom into the acridone cycle results in an increase of the fluorescence quantum yield and decay time whereas immobilization in a rigid polymer matrix (polystyrene) further extends the lifetime up to 18 ns. Large Stokes shifts and long emission decay time make this dye class an interesting platform for time-resolved imaging and sensing applications.
Analysis was performed with APCI ionization on the Advion expressionL Compact Mass Spectrometer (CMS).
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.
