Peptide purification via flash chromatography has recently been demonstrated as a viable alternative to the more standard HPLC methods currently utilized. Flash chromatography offers peptide chemists the advantage of significantly greater loading capacity reducing the overall purification time but with a compromise of decreased peak resolution. Herein we present several strategies that, when implemented, allow for very high purity peptide samples purified by flash chromatography.
Biotage Sfär columns are quality tested to ensure they meet stringent performance criteria including efficiency and peak symmetry. Each CE-marked column is built using inert, foodgrade plastics for lower extractables, cleaner fractions, and packed to provide excellent resolution.
Die Flash-Chromatographie ist die bevorzugte Reinigungsmethode für organische Stoffe, Arzneimittel und Naturstoffe. In jüngster Zeit hat sie auch die Peptidchemie erobert, verfügt sie doch über die Fähigkeit, eine Vielzahl unterschiedlicher Verbindungen effizienter zu trennen, als dies mit anderen Vorreinigungsverfahren wie z. B. dem Ausfällen (Protein-Crash) oder der Flüssig-Flüssig-Extraktion möglich ist. Zur Herstellung reiner Verbindungen können Chemiker je nach dem gewünschten Reinheitsgrad auf eine Vielzahl unterschiedlicher Variablen zurückgreifen. In diesem Whitepaper möchten wir die Faktoren erläutern, die für eine erfolgreiche Aufreinigung mithilfe der Flash-Chromatographie kontrolliert werden müssen.
Flash chromatography, a staple component of medicinal chemistry workflow, consumes a lot of organic solvent (upwards of half a million liters annually in just North America). Most, of this organic waste is incinerated off-site, liberating CO2 into the atmosphere. Because of this environmental impact, many companies are instituting requirements to reduce organic solvent waste but leaving the implementation to their chemistry departments. In this poster, we describe several proven ways to reduce solvent use without sacrificing purification efficiency.
Confidence is what you need before you separate your precious compound on a new brand of flash chromatography column. Predictably getting your compound in a pure state is what you want, and if you can get it faster and use less solvent all the better. After all, your project needs to be completed on time, and trying something new is not always appealing. In order to take your first steps toward gaining that confidence you can follow the same procedures we use here at Biotage to evaluate columns.
In this application note, we compare the amounts of a threecomponent mixture that can be separated on columns of the same size, 25 g, but with different Silica material: SNAP KP-Sil (average 50 μm irregular particles), Sfär Silica (60 μm spherical particles) and Sfär Silica HC (20 μm spherical particles) using a Biotage® Selekt instrument.
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
Synthetic chemistry is hard enough; purifying the reaction product can be even more challenging. If your reaction mixture shows you made your target but also many byproducts, what do you do? You can resynthesize using different reaction conditions, use advanced work-up procedures, or you can purify using high-performance flash chromatography.
Normal-phase flash chromatography1 has been widely adopted as the method of choice for separation of product mixtures and reaction by-products. One of the most significant developments in this area concerns the practical separation of polar molecules. Reversed-phase purification is a modification of normalphase chromatography that provides an efficient mechanism for the separation of polar compounds.
Biotage® Selekt purification platform working in tandem with improved Biotage® Sfär columns has led to a leap forward in speed of purification and a downward trend in solvent consumption. Our team has crunched the numbers, and we’re more than happy to reveal the results.
Wetting and equilibration of C18 columns for flash chromatography are best performed at the highest possible pressures/ flowrates in order to remove the air trapped inside the porous material. That will make more C18-sites activated and accessible during the chromatography run.