Avoid workflow bottlenecks by streamlining the flash purification process in your lab. This whitepaper can help you identify, reduce, or even eliminate these bottlenecks, tanking your standard operational time from hours to minutes. Learn how essential tools including TLC, a smartphone application and a simple dipping probe for fraction ID offer a novel, systematic approach to help accelerate both synthesis and purification.
Key Objectives:
Learn the key tools to setting up a streamlined purification workflow in your lab.
Review the most time-consuming processes in a routine flash purification workflow and how to avoid these bottlenecks.
Explore how modern technology, including a smartphone application, can assist with method development.
Benjamin T. Manard*, Kayron T. Rogers, Brian W. Ticknor, Shalina C. Metzger, N. Alex Zirakparvar, Benjamin D. Roach, Debra A. Bostick, and Cole R. Hexel
Abstract
The ability to directly measure uranium isotope ratios on environmental swipes has been achieved through a solution-based microextraction process and represents a significant advancement toward the development of a rapid method to analyze international nuclear safeguard samples. Here, a microextraction probe is lowered and sealed onto the swipe surface, and analytes within the sampling site (∼8 mm2) are dissolved and extracted into a flowing solvent of 2% nitric acid (HNO3). The mobilized species are subsequently directed into an inductively coupled plasma-mass spectrometer (ICP-MS) for accurate and precise isotope ratio determination. This work highlights the novelty of the sampling mechanism, particularly with the direct coupling of the microextraction probe to the ICP-MS and measurement of uranium isotope ratios. The preliminary method detection limit for the microextraction-ICP-MS method, utilizing a quadrupole-based MS, was determined to be ∼50 pg of 238U. Additionally, precise and accurate isotope ratio measurements were achieved on uranium reference materials for both the major (235U/238U) and minor (234U/238U and 236U/238U) ratios. While the present work is focused on directly measuring uranium isotopic systems on swipe surfaces for nuclear safeguards and verification applications, the benefits would extend across many applications in which direct solid sampling is sought for elemental and isotopic analysis.
Advion Interchim Scientific® Plate Express™ (Advion Interchim Scientific®, Ithaca, NY) was utilized to extract uranium from the swipe surfaces.
This webinar features several useful tools to simplify or even eliminate the sample preparation involved prior to mass spectrometry. In this educational session you will learn about several prep-free mass spec techniques that are available, and how to select which method is best for your analysis. Advion will share their perspectives on some of the new tools and protocols to minimize and simplify sample preparation. You’ll learn how to reduce your time in the lab and see your mass spec results in as little as 30 seconds!
As an attendee, you will learn more about:
How to select the best tools for sample introduction based on your compound
How new tools can reduce or eliminate sample preparation for results in <30 seconds
New workflows to maximize your time in the lab by simplifying complex processes
This webinar was presented at the 2023 LabXpo Virtual event by Lab Manager and LabX, recorded July 20, 2023.
Authors: USDA-Agricultural Research Service, Agios Pharmaceuticals, TIC Gums, United States; Universidade Federal de Lavras, Brazil
Abstract
Ethanol extract of cell mass of Serratia marcescens strain N4-5, when applied as a treatment to cucumber seed, has been shown to provide control of the oomycete soil-borne plant pathogen Pythium ultimum equivalent to that provided by a seed-treatment chemical pesticide in some soils. Two dominant compounds in this extract, prodigiosin and the serratamolide serrawetin W1, were identified based on mass and collision induced dissociation mass fragmentation spectra. An additional four compounds with M+H+ masses (487, 541, 543, and 571) consistent with serratamolides reported in the literature were also detected. Several other compounds with M+H+ masses of 488, 536, 684, 834, 906, and 908 m/z were detected in this ethanol extract inconsistently over multiple liquid chromatography coupled with tandem mass spectrometry (LC/MS–MS) runs. A purified preparation of prodigiosin provided control of damping-off of cucumber caused by P. ultimum when applied as a seed treatment while ethanol extract of cell mass of strain Tn246, a transposon-mutant-derivative of strain N4-5, did not. Strain Tn246 contained a mini-Tn5 Km insertion in a prodigiosin biosynthetic gene and was deficient in production of prodigiosin. All other compounds detected in N4-5 extract were detected in the Tn246 extract. This is the first report demonstrating that prodigiosin can control a plant disease. Other compounds in ethanol extract of strain N4-5 may contribute to disease control.
TLC/MS characterization of prodigiosin was performed using the Advion Interchim Scientific® Plate Express™ Automated TLC Plate Reader and expression® Compact Mass Spectrometer (CMS).
University of Surrey, Surrey and Borders Partnership NHS Foundation Trust, Advion, Ltd.
Abstract
Surface mass spectrometry methods can be difficult to use effectively with low cost, portable mass spectrometers. A new analytical method is presented where analytes are extracted from a sample using a solvent flushed across the surface under high pressure, separated using a chromatography column and then analysed using the Advion Interchim Scientific® expression® Compact Mass Spectrometer (CMS). In this publication, we demonstrate the efficacy of the method for the quantitative detection of cocaine and benzoylecgonine in urine and oral fluid.
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.
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.
