Welcome to the Biotage Flash Purification Blogs.

    How to Purify your Target Compound with Minimal Impurities

    February 23, 2021 at 2:05 PM / by Bob Bickler posted in Chromatography Fundamentals, purification

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    Purifying your synthetic product efficiently in high yield with minimal impurities is every chemist’s goal. At discovery-scale, flash chromatography is the go-to purification technique as it is relatively simple and effective.

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    How does gradient slope impact flash chromatography loading capacity?

    February 9, 2021 at 1:40 PM / by Bob Bickler posted in Chromatography Fundamentals, Reversed-phase, Troubleshooting and Optimization, Scale-Up

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    For many synthetic chemists the primary purification goal is to isolate as much synthetic product as possible with a minimum of 80% purity. The go-to technique for product isolation is flash purification (flash chromatography), especially for intermediates.

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    Which solvent is better for reversed-phase flash chromatography - methanol or acetonitrile?

    January 26, 2021 at 1:49 PM / by Bob Bickler posted in Chromatography Fundamentals, acetonitrile, methanol, selectivity

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    An interesting question for synthesis chemists as the need for reversed-phase flash chromatography for both intermediates and final compounds increases. Historically, normal-phase flash chromatography has been the “go-to” purification tool for synthetic intermediates. Today, however, intermediates and final products are increasingly polar. These polar reaction mixtures cause purification challenges in both separation and recovery with normal-phase (silica) thus requiring use of reversed-phase flash chromatography.

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    TLC-based vs. Generic Gradients – Which Gives the Best Results?

    January 12, 2021 at 1:40 PM / by Bob Bickler posted in Chromatography Fundamentals, Green, Method development

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    Over the course of my career, I have had terrific interactions with a multitude of chemists discussing chromatography. When it comes to flash chromatography the approaches from these chemists ranged from running generic 0-100% ethyl acetate in hexane gradients to modeled gradients based on tribal knowledge for a type of synthetic molecule to always using TLC for method development to prep HPLC. Each of these techniques are used because they provide some level of success. To me, though, if I spend time and resources synthesizing a highly valuable and unique molecule, then I want to purify the reaction mixture with the best possible method.

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    Can Flash Chromatography be Green? Part 1

    January 4, 2021 at 2:30 PM / by Bob Bickler posted in Amine, Chromatography Fundamentals, Solvents, Green, Media and Resin

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    The term “Green Chemistry” has become a major part of the science community’s lexicon. When I think about green chemistry and its relationship to flash column chromatography I think of two specific areas where it applies...

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    Can Flash Column Chromatography be Green? Part 2

    January 4, 2021 at 2:28 PM / by Bob Bickler posted in Chromatography Fundamentals, Green

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    Applying green chemistry principals to flash purification is becoming increasingly important.  In this post, I discuss ways to make flash column chromatography greener by reducing solvent use through optimization of gradient shape.

    This is a follow-on to my earlier post where I presented some greener alternatives to DCM as a solvent in flash column chromatography.

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    When should I use a “high-performance" flash chromatography column?

    December 29, 2020 at 5:00 PM / by Bob Bickler posted in Chromatography Fundamentals, efficiency

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    The evolution of flash column chromatography has brought chemists many new and exciting options for crude mixture purification. Among them are so-called high-performance flash columns or cartridges.  These high-performance purification tools are typically filled with silica or other media 15 to 30 microns in particle diameter (versus 40-63 micron for standard flash media) and provide the expectation of better separations and higher purity fractions.  That’s really enticing but how often do you need these types of columns especially since they are, of course, more expensive?  Well, that question is what I address below.

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    Why is TLC Rf important for flash column chromatography optimization?

    December 28, 2020 at 4:10 PM / by Bob Bickler posted in Chromatography Fundamentals, TLC

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    Many chemists I talk to understand that TLC data is useful for flash chromatography method development. Most also know that they should try to get their target compound to elute with an Rf between 0.15 and 0.4 by adjusting TLC solvent strength. Have you ever wondered why this is important and how Rf values impact flash chromatography results?

    In this post I will explain the relationship between TLC Rf and flash elution volumes (CV) and why having your target compound elute in the Rf range is needed.

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    Using pH to optimize reversed-phase flash chromatography separations

    December 8, 2020 at 8:51 PM / by Bob Bickler posted in Chromatography Fundamentals, Reversed-phase, Solvents, Media and Resin, Detector

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    I have previously posted on the topic of normal-phase optimization by evaluating different solvent blends or mixtures. I have also touched on reversed-phase method development as well suggesting chemists use HPLC to optimize their purification.

    In this post, I will look at the impact modifying mobile phase pH can have on reversed-phase separations.

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    Detecting the undetectable in flash column chromatography, part 2

    December 1, 2020 at 2:19 PM / by Bob Bickler posted in Chromatography Fundamentals, Troubleshooting and Optimization, Detector

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    Compound detection challenges are, for many chemists, a part of life. In a previous post I discussed how wavelength focusing can help your flash system detect and fractionate compounds with poor chromophores. However, compounds naked to UV-Vis light,  such as carbohydrates, are impossible to detect by UV when separating by liquid chromatography.

    There are some alternatives, however, and in this post I will discuss the application of evaporative light scattering detection (ELSD) to flash purification.

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    Does high performance flash column chromatography require high performance TLC for method development?

    November 25, 2020 at 4:34 PM / by Bob Bickler posted in Chromatography Fundamentals, TLC

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    Higher performance flash columns are becoming all the rage these days.   Chemists are using them for challenging as well as for routine purification.   As a result, I am often asked, "do I need high-performance TLC plates for method development?"

    In this post I will explain why the answer is no.

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    When should I add acid to my detector make-up solvent when using mass-directed flash chromatography?

    November 20, 2020 at 2:57 PM / by Bob Bickler posted in Amine, Chromatography Fundamentals, Reversed-phase, Solvents

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    Increasingly, organic and medicinal chemistry labs use mass-directed flash chromatography to isolate synthesized compounds. Mass-directed flash chromatography benefits are many, including collecting only the targeted molecule(s) in the reaction mixture. This approach simplifies compound purification since you know what you have made and it's associated mass.

    However, there are mass detection nuances that can be challenging. One of these is to know when an acid should be added to the mass detector’s make-up solvent to protonate targeted molecules. In this post, I will provide some insight on this topic.

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    Can reversed-phase chromatography compound elution volume/time be predicted?

    November 18, 2020 at 9:26 PM / by Bob Bickler posted in Chromatography Fundamentals, Reversed-phase, Method development

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    When developing reversed-phase flash chromatography methods it is important to understand the impact that a change in solvent ratio has on compound retention and, therefore, separation performance. Unlike normal-phase chromatography where you can optimize separations using TLC and a wide variety of solvents and solvent ratios, reversed-phase limits you to 3 to 4 solvents, including water, using either HPLC or small flash columns for method development. Those solvents include:

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    Invest 10 minutes on TLC and save a day of grief

    November 13, 2020 at 1:17 PM / by Bob Bickler posted in Chromatography Fundamentals, Green, TLC

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    Flash column chromatography is used by between 20 and 40 thousand organic synthesis chemists worldwide, an amazing number. For most of these chemists flash chromatography is an important part of their daily workflow but allocating time for good method development is often not considered, which can lead to less than ideal purification results.

    In this post I focus on how allocating just 10 minutes on thin-layer chromatography (TLC) for method development can save you a lot of grief later on.

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    Why is my UV baseline changing during flash column chromatography?

    November 6, 2020 at 8:15 AM / by Bob Bickler posted in Chromatography Fundamentals, Solvents, Media and Resin

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    A baseline that rises or drops when using flash chromatography with a UV detector can be a problem, especially if you are trying to collect compounds with poor detectability or that exist in low quantities.

    In this post I will talk about the causes and solutions for a rising (or even dropping) baseline.

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    How does media pore size impact small-molecule flash column chromatography?

    October 30, 2020 at 6:40 PM / by Bob Bickler posted in Chromatography Fundamentals, Troubleshooting and Optimization, purification

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    For most chemists, flash purification is a means to an end. In other words, it is a tool needed to purify and isolate one compound from a mixture of compounds so that the next reaction can occur with reduced by-product formation. Other than choosing between normal- or reversed-phase, there typically is not much thought put into cartridge selection, especially not related to stationary phase media porosity.

    For most small molecules, this approach makes sense, but for larger molecules and very lipophilic compounds, factoring for media porosity should be included.  In this post, I will discuss the impact media porosity can have on chromatographic performance.

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