Isolera™ Dalton

Here we show how flash chromatography with mass directed fractionation can be used for purifying carbohydrate derivatives such as β-D-glucosamine pentaacetate 1 as a model compound, and other UV transparent or poorly UV absorbing compounds using either normal or reversed-phase chromatography.
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In this study we synthesized two closely related amphipathic peptides and show the mass directed flash purification of non-acetylated ‘18A’ from a crude mixture containing a single residue deletion sequence ‘17A’ (-Glu residue) using the Biotage® Isolera Dalton™.
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The term “Green Chemistry” has become a major part of the science community’s lexicon. In this application note we will look at two areas for flash chromatography: 1. Replacing chlorinated solvents with those considered more environmentally friendly. 2. Reducing solvent use and waste generation with more thoughtfully applied chromatography principles.
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The UV absorption spectrum of some solvents overlaps with the product they dissolve, meaning that fraction collection processes cannot distinguish between solvent and product. Luckily, there is technology that solves this problem.
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In mass detection, the presence of ions can suppress peaks from the analytes of interest. This study concludes that Isolera™ Dalton performs successful separations even with ions present in the sample.
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Fragrance compounds, being invisible to UV, can be elusive and difficult to isolate. In this application, a four component mixture of fragrance compounds was purified using Accelerated Chromatographic Isolation™ (ACI) and identified by mass detection.
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This application note shows how the two closely related peptide structures (MW 2139 and 2267) were separated using reversed phase mass directed flash chromatography with Isolera™ Dalton.
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Mass-directed flash purification is a simple and effective technique to detect and fractionate compounds with poor UV chromophores. In this study, fatty acid methyl ester mixtures were purified by mass-directed flash purification using the Biotage Isolera™ Dalton System.
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This application note outlines how to use the Isolera system to perform a size-exclusion separation as part of a sample clean up method defined by the EPA, a key sample preparation step in environmental laboratories.
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Traditional RP-HPLC techniques not always critical because improvements in solid-phase peptide synthesis techniques over the years (microwave heating and ChemMatrix® solid phase supports) have reduced the burden on downstream purification steps. Mass directed flash purification can be a viable alternative to more expensive (and lower capacity) semi- or prep-scale RP-HPLC methods.
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In this application note we demonstrate the power of mass detection in Isolera™ Dalton by investigating the incomplete separation of two dyes, Butter yellow (Mass 225.29 g/mol) and Sudan red (Mass 278.31 g/mol), by normal phase flash purification.
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Developed by collaboration with the University of Eindhoven, European Regional Marketing and R+D, this application note describes the application of Isolera Dalton to the detection of multiply charged peptide ions prior to purification. This effectively increases the range of the Isolera Dalton mass detection, for this class of compounds and demonstrates that Isolera Dalton is a fully integrated purification instrument rather than a simple MS based analytical tool.
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In this example, Isolera™ Dalton was used to fractionate completely non-UV absorbing starting materials and products from a classic reductive amination reaction. A resin bound cyanborohydride reagent was used to greatly facilitate the work flow by eliminating the need for traditional work-up, and the mixture applied to flash chromatography purification methods. Isolera, Dalton
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Isolera™ Dalton was used to fractionate closely running nicotinic extracts from commercially available tobacco leaves. Keywords: natural, products, normal, extraction, tobacco, nicotine, nornicotine
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Biotage has developed tools for every step of the organic process, with the entire workflow in mind. This dedicated suite of products vastly expands the range of options in order to truly accelerate discoveries of new molecules for future innovations.
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Be sure you have your target compound the moment your purification is complete. The Isolera™ Dalton integrates the Biotage approach to flash chromatography with true compound identification using mass detection.
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This brochure presents the extensive range of instrumentation for flash purification from Biotage. To complement these products Biotage also offers a complete range of flash consumables, including columns in a variety of sizes packed with irregular and spherical silica, making Biotage your one-stop partner for your flash purification needs. Keywords: Isolera™ Dalton, Isolera™ Spektra, 10 Isolera™ Spektra One and Four, Isolera™ Spektra LS (Large Scale), Isolera™ Prime, Isolera™ ELSD-1080, Biotage® Flash 75/150, Biotage® Flash 400
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User report: Flash instruments. Chugai Pharmaceutical uses Biotage flash chromatography products for drug discovery research. When deciding to convert from manual open-column procedures to automated systems, they chose successive generations of Biotage products, ranging from the Flash+® packed column to the Biotage® Horizon, SP1, Isolera™ Spektra, and Isolera™ Dalton automated flash chromatography systems.
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User Report: Isolera™ Dalton, Okayama University. The laboratory of Prof. Hiroyuki Miyachi at Okayama University installed Biotage’s Isolera Dalton, an automated mass-directed purification system for flash chromatography, allowing the fractionation and collection of optical isomers. “I felt like the world’s first mass-detection automated purification system had finally arrived.”
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Leu-Enkephalin-amide (YGGFLNH2, Ca. MW = 554.65) was synthesized on the Rink amide ChemMatrix® resin at a scale of 0.5 mmol. Crude peptide (100 mg) was purified in duplicate using the Isolera Dalton equipped with a Biotage® SNAP KP-C18-HS 12g cartridge.
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The direct injection kit is intended to be used to inject samples into Isolera™ Dalton Nanolink using a syringe with a Luer-Lok tip.
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If high back pressure is detected in the makeup pump lines, and/or the High Makeup Flow Pressure dialog appears, the microfilter may be partially clogged and needs replacing. This is a step by step guide.
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Isolera™ Dalton System Care Guide
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How to Install Isolera SW 2.3
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Flash purification involves a simple liquid chromatography technique » Method development uses TLC as a way of deciding the parameters for the separation » Isocratic separations are easiest to develop, but gradient separations are more powerful » Software in the Isolera helps with conversion of an isocratic separation to a gradient » It is possible with the Spektra software to run step gradients » Loading options are dependent on the column type » SNAP offers the most flexibility » Care must be taken to choose the best loading option to get good purifications
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Preparation, How Much are You Purifying?
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The Isolera leak detection system is designed with safety in mind. Unlike vapor sensing devices with unreliable solvent vapor sensitivity, the Isolera leak detector’s sensitive, solvent-resistant refractive index (RI) monitor rapidly signals the Isolera flash system to stop pumping if any liquid, volatile or non-volatile, is detected thus protecting the chemist, system, and lab from potential hazards.
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Improvements in solid phase peptide synthesis strategies and development of resin linkages susceptible to low acid cleavage conditions has enabled synthesis of long peptides while keeping the protecting groups intact. This strategy is now used for the preparation of chemically synthesized proteins, wherein shorter peptide fragments are ligated together. They are also found in the synthesis of peptide macrocycles that utilize head-to-tail cyclization strategies. Although linear synthesis of protected peptides is generally straightforward, purification of these compounds using traditional reversed phase methods is quite challenging. Herein we describe the use of normal phase chromatography for purification of fully protected peptides.
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Natural product chemistry deals with discovering the previously unknown in nature. Compounds found in nature are typically found in low quantity and thus extractions are needed to isolate certain compounds classes or at least compounds with similar solubility.
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For most organic and natural product chemists flash chromatography is a necessary part of their research. As such, many chemists need quick isolation of at least one desired component from a crude mixture in relatively high yield and purity. This need for speed, purity, and yield pits these desires against each other as you can typically optimize on only two of the three goals. In this poster, we will describe some techniques that help chemists optimize flash purification and maximize speed, yield, and purity.
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Although capable of very high resolution, RP-HPLC is often limited by low column loading capacity, therefore demanding a significant time investment for peptide purification. As an alternative strategy, reversed-phase flash chromatography can also be used to purify synthetic peptides. The larger particle size used in flash column chromatography enables much larger loading capacity, thereby significantly reducing the time required for peptide purification.
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When it comes to polar organic compound purification, many chemists turn to normal-phase flash chromatography often utilizing dichloromethane and methanol as the eluting solvents. While this can work, it often can be challenging to optimize due to methanol’s high polarity and protic chemistry.
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As reversed-phase flash chromatography gains traction in medicinal chemistry labs the need to monitor its cost and safety are becoming more important. Commonly used reversed-phase solvents typically include water with an organic solvent such as methanol or acetonitrile – each have advantages and disadvantages.
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For those chemists performing organic synthesis, reaction mixture purification by flash column chromatography is an integral and necessary part of the synthesis process. However, flash chromatography consumes large volumes of solvent which either needs to be recycled or disposed. ACS 2016.
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Reversed-phase chromatography is typically used when you need to separate several milligrams of relatively polar compounds that either are not soluble in normal-phase solvents or are not compatible with bare silica because they react, stick, or both. If you are currently using reversed-phase at preparative scale, such as flash chromatography, you know the mobile phase limitations – water with either methanol, acetonitrile, or THF. As with normal-phase flash chromatography, when it comes time to purify you want your crude sample fully solubilized in the weakest possible solvent at the highest possible concentration. ACS 2016
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DMSO and DMF are suitable injection solvents for reversed-phase flash purification. DMSO shows it can be loaded in larger volumes (up to 0.05 mL/g of C18 media or 3.5% of a column volume) without affecting chromatographic separations or carrying compounds with it.
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New, higher default flow rates enabled by Isolera ACI systems decreases purification times compared to other systems’ and cartridges’ default flow rate settings without sacrificing separation efficiency. Incorporation of gradient optimization by converting a linear gradient to a step gradient further reduces purification time and solvent use while maintaining separation integrity for a target molecule.
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In this poster we demonstrate real-time detection and identification of peptides during reversed-phase flash chromatography with the new Biotage® Isolera™ Dalton mass-directed flash purification system. Presented at ACS Spring 2014.
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In this poster we will show how using reversed-phase flash chromatography with the new Biotage® Isolera™ Dalton mass-directed flash purification system provides real-time simultaneous separation, detection, and identification of lipids. Presented at ACS Spring 2014.
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In order to obtain a fast synthesis of an N-unprotected tripeptide, microwave (mw) assisted deprotection, coupling and partial deprotection were applied starting with a purified protected dipeptide [Fig. 1]. The three consecutive mw assisted steps were performed without any intermediate purifications and were accomplished within some 30 minutes. The resulting crude mixture containing the Boc deprotected tripeptide ester was shown to be quite complex [Fig. 2]. The crude product was subjected to automated reversed phase flash chromatography, followed by automated fraction pooling and “thin film evaporation”. The total time for the entire procedure was 1 h 20 min. The purity of the product was > 99%.
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The tasks carried out during the maintenance visit shall include servicing the listed components and a final check which is based on factory acceptance and qualifications tests. These two procedures constitute a Biotage Preventive Maintenance (PM) Plan.
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Using the Biotage automated workflow, models have shown that the time taken to produce a target molecule can be reduced by up to eighty percent, allowing project delivery timescales to be reduced accordingly.
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Traditional approaches to compound purification involving chromatography utilize large volumes of relatively toxic and expensive solvents, and significantly contribute to the environmental footprint of organizations involved in molecular research. Current directives for greener chemistry have put pressure on organizations to reduce the environmental impact of their work.
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This document outlines service products specific to Isolera™ Dalton. For further information on Biotage service products please see “Biotage Service Agreements” brochure (PPS330).
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With running costs and time a concern in most labs, ACI™ now promises to take users to the end point even faster and even more efficiently.
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The mass-directed flash purification system Isolera™ Dalton produces its final mass spectrum by compiling data from hundreds of individual mass spectra in real time.
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Isolera Dalton identifies compounds by mass in real time during flash separation, leading to greater confidence in purification, higher purity, greater recovery, and a significant saving in time. Reduce or eliminate the need for time consuming off-line mass analysis during development programs, increase throughput and effectiveness for both chemists and analytical groups.
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The Isolera leak detection system is designed with safety in mind. Unlike vapor sensing devices with unreliable solvent vapor sensitivity, the Isolera leak detector’s sensitive, solvent-resistant refractive index (RI) monitor rapidly signals the Isolera flash system to stop pumping if any liquid, volatile or non-volatile, is detected thus protecting the chemist, system, and lab from potential hazards.
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